Registration Dossier
Registration Dossier
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EC number: 231-272-0 | CAS number: 7473-98-5
Endpoint summary
Administrative data
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Effects on fertility
Description of key information
The reproductive toxicity of the registered substance was determined in an Extended One Generation Reproductive Toxicity study using read-across to a analogue substance.
Administration of the source substance by once daily (or twice daily 3 hours apart on some occasions) oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no test item-related deaths. There were occasional observations of decreased activity, abnormal, uncoordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose level-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body weight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirmed by overall ambulation values, or any associated clinical findings, detailed functional observations or brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 males only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels (up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertrophy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney weights were seen in both sexes. In neurological investigations, there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions. There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1 adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the in-life and pathological findings were considered not to be adverse.
Link to relevant study records
- Endpoint:
- extended one-generation reproductive toxicity - with F2 generation and developmental neurotoxicity (Cohorts 1A, 1B with extension, 2A and 2B)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Remarks:
- Information on target substance.
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
See attachment
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See attachment
3. ANALOGUE APPROACH JUSTIFICATION
See attachment
4. DATA MATRIX
See attachment - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 424. Neurotoxicity Study in Rodents
- Version / remarks:
- 21 July 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 443 (Extended One-Generation Reproductive Toxicity Study)
- Version / remarks:
- 25 June 2018
- Deviations:
- no
- Limit test:
- no
- Justification for study design:
- ECHA decisions on testing proposals:
Target substance requirement:
Extended one-generation reproductive toxicity study (Annex X, Section 8.7.3.; test method: OECD TG 443) in rats, oral (gavage) route specified as follows:
- Ten weeks premating exposure duration for the parental (P0) generation;
- Dose level setting shall aim to induce systemic toxicity at the highest dose level;
- Cohort 1A (Reproductive toxicity);
- Cohort 1B (Reproductive toxicity) without extension to mate the Cohort 1B animals to produce the F2 generation.
Source substance requirement:
Additional data generated from the extension to mate the Cohort 1B to produce the F2 generation and also the Cohorts 2A and 2B (developmental neurotoxicity) was a requirement of the decision on a testing proposal for the source Hydroxycyclohexyl phenyl ketone (TPE-D-2114510002-76-01/F) and that this has been included for openness and completeness of reporting of the actual experimental study performed. - Species:
- rat
- Strain:
- Wistar
- Remarks:
- Han Wistar (Crl:WI(Han))
- Details on species / strain selection:
- The Han Wistar rat was chosen as the animal model for this study as it is an accepted rodent species for nonclinical toxicity testing by regulatory agencies, it is also the preferred model for this OECD guideline study.
- Sex:
- male/female
- Route of administration:
- oral: gavage
- Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- Control
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Dose / conc.:
- 900 mg/kg bw/day (nominal)
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- At 900 mg/kg/day, decreased activity was observed for one male on Day 2 and abnormal, unco
ordinated gait for 3 males and 4 females between Days 2 to 4. Decreased activity, abnormal, uncoordinated gait or low carriage were occasionally noted for a further 8 females between Days 5 to 70 and decreased activity on one occasion for each of 2 females during lactation. These findings were considered to be test item-related.
Salivation and/or ploughing (also a chewing action for one female at 900 mg/kg/day) were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, on multiple days throughout the dosing period. The incidence of these findings was dose level-related.
Animal 4515F (900 mg/kg/day) had abnormal clincal observations of irregular respiratory rate, hunched posture, erect fur, cold to touch, prominent backbone, abnormal gait, decreased activity and red discharge from vagina the day after parturition (LD 1) but these mostly resolved before scheduled termination. The clinical observations were considered to be likely due to the trauma of giving birth rather than being test item-related.
Animal 4525F (900 mg/kg/day) had abnormal clincal observations of irregular/decreased respiratory rate, erect fur, eyes partly closed, decreased activity up to 2-4 hours postdose on LD 7 only. No
similar findings were observed for this animal during the dosing period and as a cause could not be
determined, these findings were likely related to the dosing procedure. - Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- None of the unscheduled deaths were considered to be test item-related.
On Study Day 40, approximately 3 hours after dosing, Animal 4510F given 900 mg/kg/day was found
with clinical signs of irregular respiratory rate, hunched posture, partly closed eyes, erect fur, unc
oordinated and abnormal gait and decreased activity and was euthanised. There were no findings
at necropsy or microscopically to suggest the likely cause of death. This animal had not had any
abnormal clinical signs before this (apart from salivation and ploughing) and had consistently gained
body weight.
Animals 2512F and 4502F given 100 or 900 mg/kg/day respectively, were euthanised on LD 1 or LD
0 and due to the very young age of the pups, they were also euthanised. Animal 2512F had clinical
signs of irregular respiratory rate, hunched posture, pale skin and cold to touch, erect fur, uncoor
dinated and abnormal gait in the afternoon of LD 1. Animal 4502F was found subdued, cold to touch,
prostrate/lying on side with decreased respiratory rate approximately 1 hour after dosing on LD 0. Th
ere were no findings at necropsy or microscopically to suggest the likely cause of death and both an
imals had not had any abnormal clinical signs before this (apart from salivation and ploughing for 4
502F) and had consistently gained body weight. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See Tables 1-4.
For males given 900 mg/kg/day, there was slightly lower body weight gain over the dosing period resulting in male body weight being 3.6% lower than the concurrent control mean value by Day 134. For females given ≥ 100 mg/kg/day, a dose level-related higher body weight gain of up to 10.3% at 900 mg/kg/day compared to controls was noted during the pre-pairing period and this slightly higher body weight was generally maintained throughout gestation and lactation.
There was no test item-related effect on male body weight at 100 or 300 mg/kg/day. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- There were occasional slightly higher food consumption values of up to 14.4% of controls noted for
males at 300 or 900 mg/kg/day up to Day 106. At ≥100 mg/kg/day for males from Days 106 to 134,
there was higher food consumption of up to 17.5%. For females given ≥100 mg/kg/day, there was an
overall dose level-related higher food consumption of up to 20.6% of control values during the prepairing
period, and during gestation and lactation, food consumption was higher for females by up to
13.2% at 300 or 900 mg/kg/day only. - Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Haematology
See tables 18-19.
Coagulation
See tables 15-16.
All differences in haematology or coagulation parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control values and/or were of a magnitude of variation commonly observed in rats. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 12-13.
At ≥300 mg/kg/day for males and ≥100 mg/kg/day for females, triglycerides were lower than controls by up to 0.75-fold, and for males only at ≥300 mg/kg/day, cholesterol higher by up to 1.30-fold. - Endocrine findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of
necropsy in F0 adult males given 300 or 900 mg/kg/day.
There were no test item-related effects for F0 adult males given 100 mg/kg/day or F0 adult females gi
ven up to 900 mg/kg/day or for culled pups on PND 4 or unselected pups on PND 21.
There were no test item-related changes to T4 levels at up to 900 mg/kg/day for samples taken on the
morning of necropsy for the F0 generation animals, for culled pups on PND 4 or unselected pups on
PND 21.
The differences in these parameters were considered not to be test item-related based on their small
magnitude, inconsistent direction, absence of a dose response, general overlap of individual values
with the range of control and/or were of a magnitude of variation commonly observed in rats. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See tables 27-28.
All differences in urine parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats. - Immunological findings:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Test item-related Microscopic Findings
In the liver, there was diffuse centrilobular hepatocyte hypertrophy of minimal to mild severity. For
males only, this was seen in all dosage groups, whereas for females this was only identified in ani
mals dosed at 900 mg/kg/day. Although this was also found in one control animal (M1021), the
incidence was higher in animals treated with the test item and followed a dose-related distribution.
This finding was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in f
emales dosed at 900 mg/kg/day.
The higher liver weights recorded in animals given the test item, were correlated to the histopatholo
gic findings of centrilobular hypertrophy. This correlation was evident in the males dosed at ≥ 100 mg/
kg/day and females dosed at 900 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal
severity. This was characterised by an increase in the number of goblet cells. For males only, this was
seen in at ≥ 300 mg/kg/day, whereas for females this was only identified in animals dosed at 900 mg/
kg/day. Although this was also found in one control animal (M1013), the incidence was convincingly
higher in animals treated with the test item at ≥ 300 mg/kg/day and showed a dose-related distribut
ion. This finding was therefore considered test item-related in males dosed at ≥ 300 mg/kg/day and in
females dosed at 900 mg/kg/day.
Additional Microscopic Findings:
In the spleen, there was a slightly higher incidence of increased hematopoietic cells with a minimal
severity, primarily in males given the test item at a doses of 100 mg/kg/day and 900 mg/kg/day. A
slightly higher incidence was seen in females dosed at 900 mg/kg/day only. However, due to the
overall low incidences and because this is a common background finding, it was considered not test i
tem-related.
Other microscopic findings were of the nature commonly observed in this strain and age of rat, or
occurred at a similar incidence in control and treated animals, and, therefore, were considered not to
be test item-related. - Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- There was no effect on the number or length of estrous cycles during the 2 weeks before pairing or during the mating period at up to 900 mg/kg/day, compared with controls.
On the morning of necropsy, while the majority of females in all groups were still in diestrus, a few in each group had come back into estrus. - Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- There were no test item-related effects on the percentage of motile sperm, progressively motile sperm or straight line velocity at up to 900 mg/kg/day.
F0 control male 1020M control was paired with 1520F which was found not to be pregnant at scheduled necropsy. The sperm parameters for male 1020M were within the normal ranges.
F0 Animal 2011M given 100 mg/kg/day had a very low sperm motility of 2% at necropsy, however, the female it had been paired with 2511F, became pregnant and littered successfully.
At 900 mg/kg/day, F0 Control 1024M was paired with 1524F, and 4010M was paired with 4511F. These females failed to produce litters, however, the sperm parameters for the males were within the normal ranges.
Sperm Count and Morphological Analysis:
There were no test item-related effects on sperm count or morphology at up to 900 mg/kg/day.
Spermatid Count:
There was no test item-related effect on the number of spermatids at up to 900 mg/kg/day. - Reproductive performance:
- no effects observed
- Description (incidence and severity):
- There were no test item-related effects on male or female mating performance or fertility, or duration of gestation in females, at up to 900 mg/kg/day.
F0 control 1520F was found not to be pregnant at scheduled necropsy.
F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early resorption) and 4511F was not pregnant.
F0 Animals 1501F, 1502F, 1506F, 1518F, 1521F (control), 2517F (100 mg/kg/day), 350F2 (300 mg/kg/day) and 4523F (900 mg/kg/day) did not have a positive mating sign during pairing and therefore gave birth before their expected GD 21.
See table 20, table 23, and table 25. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- body weight and weight gain
- food consumption and compound intake
- clinical biochemistry
- organ weights and organ / body weight ratios
- histopathology: non-neoplastic
- reproductive function (oestrous cycle)
- reproductive function (sperm measures)
- reproductive performance
- Critical effects observed:
- no
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- Salivation and/or ploughing were noted for all animals (except Surplus Animal 3650F) given ≥300 mg/kg/day and for a few animals given 100 mg/kg/day, on several days throughout the dosing period. The incidence of these findings was dose level-related.
Surplus Animal 4146M had erect fur on Day 5 postdose and 4147M had irregular respiratory rate, erect fur, eyes partly closed and abnormal gait on Day 4 postdose. Both animals (given 900 mg/kg/day) did not have any similar findings during the dosing period and as a cause could not be determined, these abnormal findings were likely related to the dosing procedure.
For all generations, the other clinical observations recorded were considered not to be test item-related as they were of low incidence, are commonly seen during gestation or lactation, were also seen in some control animals or were not dose level-related. - Mortality / viability:
- mortality observed, non-treatment-related
- Description (incidence and severity):
- Animal 3608F given 300 mg/kg/day had convulsions and died within minutes of receiving its first dose
on Day 1. Grossly, the lungs had pale discoloration and abnormal consistency and the trachea had fr
othy pale content. Microscopically, there was multifocal mixed cell infiltration of the alveolar space of
the lung which was considered to correlate with the gross findings in the lung. Overall, these findings
were considered to have been related to the dosing procedure and were therefore considered to be
the cause of death.
Control Animal 1628F was euthanised on LD 1 because all pups in the litter died. At necropsy, a
pale pancreas was noted but no microscopic findings could be correlated and no cause of death w
as identified. The animal had not had any abnormal clinical signs before this and had consistently g
ained body weight.
Animal 4634F was found dead in the afternoon of LD 18. At necropsy, in the abdominal cavity, there
was dark red fluid accumulation and a soft red mass adherent to the stomach. Microscopically this
was correlated with a well demarcated hematocyst of marked severity in relation to hepatic tissue. Thi
s finding was considered to be the animal’s cause of death. As it’s pups had not been weaned, they w
ere also euthanised. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 5-11.
For animals in Cohorts 1A and 1B, there was a general trend of slightly lower body weight gain (up to
7.0%) for males at 900 mg/kg/day compared to controls, but no test item related-effects for males at 100 or 300 mg/kg/day or females at ≥100 mg/kg/day during pre-pairing, gestation or lactation.
In Cohort 2A and for Surplus animals, body weight gain was variable and did not follow any trend. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- There were occasional higher food consumption values of up to 19.6% of controls noted for males
at 300 or 900 mg/kg/day, in some F1 generation cohorts. There was a general trend of higher food
consumption for females of up to 22.4% higher at 300 or 900 mg/kg/day but during gestation and la
ctation female food consumption was similar to controls. - Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See table 20.
All differences in haematology or coagulation parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control values and/or were of a magnitude of variation commonly observed in rats. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- See table 14.
At ≥100 mg/kg/day for males and females, triglycerides were lower than controls by up to 0.62-fold,
and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold.
For both generations, all other differences in clinical chemistry parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See table 29 .
All differences in urine parameters were considered not to be test item-related
based on their small magnitude, inconsistent direction, absence of a dose response, general overla
p of individual values with the range of control and/or were of a magnitude of variation commonly ob
served in rats. - Sexual maturation:
- no effects observed
- Description (incidence and severity):
- There was considered to be no effect on sexual maturation for F1 males or females at up to 900 mg/kg/day.
- Anogenital distance (AGD):
- no effects observed
- Description (incidence and severity):
- There was no effect on anogenital distance or normalised anogenital distance for males or females at up to 900 mg/kg/day.
- Nipple retention in male pups:
- no effects observed
- Description (incidence and severity):
- On PND 13, there was no effect on nipple retention in male pups i.e. no nipples were present, at dose levels up to 900 mg/kg/day.
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- F1 Cohort 1A
Test item-related Organ Weight Differences:
Liver weights in animals dosed at ≥ 300 mg/kg/day were statistically significantly higher than the controls and showed a dose-related response. In males, liver weights were up to ~24% higher and in females up to ~29% higher (relative to body weight at 900 mg/kg/day). This was therefore considered test item-related.
Kidney weights in animals dosed at ≥ 300 mg/kg/day in males and at ≥ 100 mg/kg/day in females, were statistically significantly higher than the controls and showed a dose related response. In males, kidney weights were up to ~23% higher and in females up to ~16% higher (relative to body weight at 900 mg/kg/day). Although this was not correlated to any microscopic findings, this was considered test item-related in view of the similar weight differences recorded in the F0 and F1 cohort 1B animals.
Additional Organ Weight Differences:
Testis weights relative to body weight were statistically significantly higher by ~11% only at 900 mg/kg/day compared to the controls. However, males dosed at 900 mg/kg/day had a lower terminal body weight and in view of the minor weight difference and no microscopic findings that could be correlated. This was therefore considered incidental.
Ovary weights were statistically significantly higher by ~19% (relative to body weight at 300 mg/kg/day). However, no histologic findings could be correlated to this finding and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Thyroid/parathyroid gland weights were statistically significantly higher by ~21% (relative to body weight) in female animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, thyroid hormone was not increased in a noteworthy fashion, and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Seminal vesicle weights were statistically significantly higher by ~22% (relative to body weight) in animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, so this was considered not test item-related.
There were individual organ weight values that were different from their respective controls. There were, however, no patterns or correlating data to suggest these values were test item-related.
F1 Cohort 1B
Test item-related Organ Weight Differences:
Liver weights in animals dosed at ≥ 300 mg/kg/day were statistically significantly higher than the co
ntrols and showed a dose-related response. In males, liver weights were up to ~24% higher and in
females up to ~29% higher (relative to body weight at 900 mg/kg/day). This was therefore considered
test item-related.
Kidney weights in animals dosed at ≥ 300 mg/kg/day in males and at ≥ 100 mg/kg/day in females,
were statistically significantly higher than the controls and showed a dose related response. In males,
kidney weights were up to ~23% higher and in females up to ~16% higher (relative to body weight
at 900 mg/kg/day). Although this was not correlated to any microscopic findings, this was considere
d test item-related in view of the similar weight differences recorded in the F0 and F1 cohort 1B an
imals.
Additional Organ Weight Differences:
Testis weights relative to body weight were statistically significantly higher by ~11% only at 900 mg/
kg/day compared to the controls. However, males dosed at 900 mg/kg/day had a lower terminal b
ody weight and in view of the minor weight difference and no microscopic findings that could be corr
elated. This was therefore considered incidental.
Ovary weights were statistically significantly higher by ~19% (relative to body weight at 300 mg/kg/
day). However, no histologic findings could be correlated to this finding and no similar differences
were found in any other generation or cohort of animals, so this was considered incidental.
Thyroid/parathyroid gland weights were statistically significantly higher by ~21% (relative to body
weight) in female animals dosed at 900 mg/kg/day, compared to controls. However, no histologic fi
ndings could be correlated to this finding, thyroid hormone was not increased in a noteworthy fashio
n, and no similar differences were found in any other generation or cohort of animals, so this was c
onsidered incidental.
Seminal vesicle weights were statistically significantly higher by ~22% (relative to body weight) in
animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be co
rrelated to this finding, so this was considered not test item-related.
(Cohort 2A, 2B, F1 unselected pups, F2 unselected pups):
There were no test item-related organ weight differences. - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- Scheduled Euthanasia Animals - F1 Animals:
There were no test item-related gross findings.
Gross findings observed were of the nature commonly observed in this strain and age of rat, or occurred at a similar incidence in control and treated animals, and, therefore, were considered not to be test item-related.
Scheduled Euthanasia Animals (F1 Unselected pups):
There were no test item-related gross pathology findings.
The only abnormality noted was a dark red focus on the thymus in one male pup. Although the parents of these pups had been given Omnirad 184 at a dose of 900 mg/kg/day, this finding was considered not to be test item-related because it is a common agonal change. - Histopathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- (Cohort 1A)
In the liver, there was diffuse centrilobular hypertrophy of minimal to mild severity. For males only,
this was seen in all dosage groups, whereas for females this was only identified in animals dosed at
≥ 300 mg/kg/day. The incidence followed a dose-related distribution and was therefore considered
test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at ≥ 300 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184 were correlated to the histopat
hologic findings of centrilobular hypertrophy. This correlation was evident in males dosed at ≥ 100 mg/
kg/day and females at ≥ 300 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal seve
rity. This was characterised by an increase in the number of goblet cells. For both females and male
s, this was seen at ≥ 300 mg/kg/day. Although at a low incidence, this finding was only seen in anim
als treated with the target substance and in view of similar findings seen in the F0 generation, this finding was
considered test item-related in males and females dosed at ≥ 300 mg/kg/day.
Additional Microscopic Findings:
In the kidneys, there was a higher incidence of basophilic tubules with a minimal to mild severity, in
male animals given the target substance at a dose of 900 mg/kg/day. This was however a small difference
and although the males from F0 and F1 cohort 1B showed a similar difference, it was even less appa
rent at the highest dose level (900 mg/kg/day). Due to the fact that this lesion is a common backgro
und finding and occurred at a low severity, this was considered incidental.
In the liver of three animals given the target substance (two females dosed at 100 mg/kg/day and one female
dosed at 300 mg/kg/day), there was focal necrosis with subacute inflammation in the medial lobe
parenchyma, with a mild severity. Due to the low incidence, this finding was considered not test item related.
(Cohort 1B)
In the liver, there was diffuse centrilobular hypertrophy of minimal to mild severity. For males only, t
his was seen in all dosage groups, whereas for females this was only identified in animals dosed at
≥ 300 mg/kg/day. The overall incidence followed a dose related distribution and was therefore consid
ered test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at ≥ 300 mg/kg/day.
The higher liver weights recorded in animals given the target substance, was correlated to the histop
athologic findings of centrilobular hypertrophy. This correlation was evident in males dosed at ≥ 100
mg/kg/day and females at ≥ 300 mg/kg/day. There were no test item-related microscopic findings in
the reproductive tissues.
(F1 Cohort 2A and 2B)
There were no test item-related microscopic findings. Microscopic findings occurred at similar incid
ences in control and treated animals, and, therefore, were considered not to be test item-related. - Other effects:
- no effects observed
- Description (incidence and severity):
- Acoustic startle:
There were no test item-related effects on maximum response amplitude, average response amplitude or time to maximum response in males or females at up to 900 mg/kg/day.
Estrous Cycles:
The number of days after vaginal opening before an estrus occurred was comparable with controls at all test item dose levels. There was no effect on the number or length of estrous cycles during the 2 weeks before necropsy at up to 900 mg/kg/day, compared with controls.
There was no effect on the number or length of estrous cycles during the 2 weeks before pairing or during the mating period at up to 900 mg/kg/day, compared with controls.
Mating Performance, Fertility and Duration of Gestation:
The following Cohort 1B females were euthanised on GD 24 as they had failed to produce litters and were found not to be pregnant; 1629F (control), 2624F, 2627F, 2628F and 2637F (100 mg/kg/day) and 3629F (300 mg/kg/day).
See table 22, table 24, and table 26
Pre- and Post-Implantation Loss:
There were no test item-related effects on number of corpora lutea, implant counts, pre- or post-implantation loss or total number of pups born at up to 900 mg/kg/day. - Behaviour (functional findings):
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test item-related effects on qualitative or quantitative functional observational para
meters at up to 900 mg/kg/day. - Developmental immunotoxicity:
- no effects observed
- Description (incidence and severity):
- Splenic Immunophenotyping Evaluation:
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the immune cell populations in either the F0 adults rats analysed following administration of Omnirad 184 by oral gavage, or their resulting offspring. - Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- food consumption and compound intake
- clinical biochemistry
- organ weights and organ / body weight ratios
- histopathology: non-neoplastic
- Critical effects observed:
- no
- Clinical signs:
- not examined
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- There were no unscheduled deaths for F2 generation animals.
- Body weight and weight changes:
- not examined
- Food consumption and compound intake (if feeding study):
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- no effects observed
- Nipple retention in male pups:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- There were no test item-related organ weight differences.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no test item-related gross pathology findings.
Abnormalities noted at necropsy included a dark red focus on the thymus in one female pup. Although the parents of these pups had been given the target substance at a dose of 100 mg/kg/day, this finding was considered not to be test item-related because it is a common agonal change. In the liver of one male and three female pups whose parents had been treated with the target substance at a dose of 100 mg/kg/day there was mottled discoloration, abnormal appearance and a pale nodule on the medial lobe. Due to the low incidence and because no other animals were found with these gross lesions, they were considered not test item-related. - Histopathological findings:
- not examined
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- no effects observed
- Description (incidence and severity):
- Splenic Immunophenotyping Evaluation:
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the immune cell populations in either the F0 adults rats analysed following administration of Omnirad 184 by oral gavage, or their resulting offspring. - Key result
- Dose descriptor:
- NOAEL
- Generation:
- F2
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- viability
- sexual maturation
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Lowest effective dose / conc.:
- 900 mg/kg bw/day
- Treatment related:
- no
- Conclusions:
- The administration of the target substance once daily (or twice daily 3 hours apart on some occasions) via
oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no
test item-related deaths. There were occasional observations of decreased activity, abnormal, un
coordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose le
vel-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body we
ight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/
day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. Ther
e were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and
F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity
was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirme
d by overall ambulation values, or any associated clinical findings, detailed functional observations or
brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/
kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 male
s only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels
(up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/
kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values
or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target
organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertro
phy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney
weights were seen in both sexes. In neurological investigations, there were no test item-related brain
weight differences, gross or histological findings or effects on the brain dimensions. There were no
histological findings in the reproductive tissues of the females that had not been observed to litter tha
t would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1
adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the i
n-life and pathological findings were considered not to be adverse. For F1 and F2 developmental para
meters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on
the pup numbers, survival or development. - Executive summary:
Read-across approach:
The methodology used for choosing potential read-across source substances for this current assessment was two-fold:
- Identify substances that have a similar molecular structure to the target substance, for example, having common functional groups as well as likely having structurally-similar degradation products, and
- Identify which of these source substances have the necessary toxicology data to fulfil the data gap.
The substance finally selected for the read-across source met the scientific hypothesis and comparison criteria below:
- The target and source substance are small molecules with common functional groups which will influence their toxicokinetic behaviour. The predicted metabolic route and hence breakdown products within the body are common for both substances.
- A comparison of existing mammalian (rat) toxicity data shows that both substances were of low toxicity, with both having high values for No Observed Adverse Effect Levels (NOAEL) for repeated dose toxicity studies in rats. In particular, no developmental toxicity was observed in either substance at the dose levels tested.
- The intended use of both substances is as Photoinitiators which means high commonality in the manufacture, formulation and use. Thus the potential for (accidental) human and environmental exposure is the same for both substances.
It is therefore concluded that the target substance will have the same reproductive toxicity outcomes as that of the source substance for an Extended One Generation Reproductive Toxicity (EOGRT) Study.
Executive summary of the study:
The objective of this study was to determine the potential toxicity of the test item, when given by oral gavage to adult rats and their offspring. This study was designed to provide an evaluation of reproduction and development as well as a thorough evaluation of systemic toxicity in pregnant and lactating females and their offspring. Detailed examination of key developmental endpoints, such as offspring viability, neonatal health, developmental status at birth, physical and functional development until adulthood, nervous and immune system development was expected to identify any specific target organs in the offspring. In addition, the study provided information about the effects of the test item on the integrity and performance of the adult male and female reproductive systems and a neurobehavioural assessment of the F1 generation.
The study design was as follows:
Text Table 1 Experimental Design – F0 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
25
25
2
Test item
100
10
10
10.03
100
25
25
3
Test item
300
10
30
30.09
301
25
25
4
Test item
900
10
90
90.27
903
25
25
a Dose volume was based on the most recent body weight measurement.
Control/vehicle = 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 2 Experimental Design – F1 Unselected PND (Post Natal Day) 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 3 Experimental Design – Cohort 1A (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Test item
100
10
10
10.03
100
20
20
3
Test item 300
10
30
30.09
301
20
21d
4
Test item
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and
450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
d One female given 300 mg/kg/day died within minutes of receiving its first dose on Day 1, and another female
was added to the group.Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 4 Experimental Design – Surplus F1 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Test item
100
10
10
10.03
100
10
10
3
Test item
300
10
30
30.09
301
10
10
4
Test item
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 5 Experimental Design – Cohort 1B (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Test item
100
10
10
10.03
100
20
20
3
Test item
300
10
30
30.09
301
20
20
4
Test item
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 6 Experimental Design – F2 Unselected PND 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 7 Experimental Design – Cohort 2A (Neurobehavioural)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Test item
100
10
10
10.03
100
10
10
3
Test item
300
10
30
30.09
301
10
10
4
Test item
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 8 Experimental Design – Cohort 2B (Neuropathology)a
Group No.
Number of Animals
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Cohort 2B were not dosed as they were terminated on PND 21/22.
In Week 17 of the study (F0 dosing and the start of F1 dosing), the 900 mg/kg/day dosing formulation could not be pulled up into the gavage tube being used for newly weaned pups, due to an unknown viscosity property, resulting in some subsets of F0 and/or F1 animals either not being dosed or being given the 300 mg/kg/day formulation either once or twice daily instead (refer to section 4.6.1 for details). From 19 Aug 2021, Group 4 F1 animals were dosed twice daily 3 hours apart at 450 mg/kg (equivalent to 900 mg/kg/day) until they were old enough for the next size gavage tube on 30 Aug 2021. Although there were occasions when animals were not dosed or received a dose level lower than that required by the study protocol, there was considered to be no overall impact on the results or study outcome due to the length of time that each generation was administered the test item dose levels correctly.
The following parameters and end points were evaluated in this study: clinical observations, body weights, food consumption, estrous cycles, mating performance, fertility indices, duration of gestation and overall litter performance, litter survival indices, litter and pup weights, pre-weaning physical development of F1 pups, assessments of sexual maturation of F1 animals, clinical pathology parameters (haematology, coagulation, clinical chemistry, and urinalysis), thyroid stimulating hormone (TSH) and thyroxine (T4) analysis, gross necropsy findings, splenic immunophenotyping analysis, organ weights, sperm evaluation, ovarian follicle counts and histopathological examinations.
None of the unscheduled deaths in any generation were considered to be test item-related.
For the F0 generation at 900 mg/kg/day, test item-related clinical observations of decreased activity, abnormal/uncoordinated gait and low carriage were occasionally seen for males and/or females before pairing, and decreased activity only during lactation. For F0 and F1 generations, salivation and/or ploughing were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, throughout the dosing periods. F0 and F1 males given 900 mg/kg/day had slightly lower body weight gain, and F0 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day.
There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development.
For males at 900 mg/kg/day, during the last 20 minutes of the 1 hour session, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this, the cause of this apparent reduction in motor activity could not be determined.
For F0 males given ≥300 mg/kg/day and F0 females given ≥100 mg/kg/day, triglycerides were lower by up to 0.75-fold, and for males only, cholesterol higher by up to 1.3-fold. For F1 animals at ≥100 mg/kg/day, triglycerides were lower than controls by up to 0.62-fold, and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold. There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day.
In the F0 animals dosed from 10 weeks prior to mating and continuing throughout mating and gestation until the females were at least LD 21, microscopically, centrilobular hypertrophy in the liver was considered test item-related in all dose groups in males but only at 900 mg/kg/day in females and mucosal hyperplasia of the glandular region, appearing as an increased number of goblet cells, was considered test item-related at ≥ 300 mg/kg/day in males and only at 900 mg/kg/day in females. Higher liver and kidney weights were seen in both sexes at all dose levels.
In the F1 cohort 1A animals, dosed from PND 21 until at least PND 90, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. In the stomach, there was mucosal hyperplasia of the glandular regions, appearing as an increased number of goblet cells. This finding was considered test item-related in males and females at ≥ 300 mg/kg/day. Liver weights were higher at ≥ 300 mg/kg/day in males and females. In addition, higher kidney weights were seen in females at all dose levels and at ≥ 300 mg/kg/day in males.
In the F1 cohort 1B animals, that were used to produce an F2 generation, and that were dosed from PND 21 until at least LD 21, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. Liver weights were higher at all dose levels in males and only at 900 mg/kg/day in females. In addition, higher kidney weights were seen in animals dosed at 900 mg/kg/day. Stomachs were not assessed microscopically in these animals. There were no test item-related organ weight differences in the female reproductive tissues.
There were no test item-related organ weight differences in a limited list of tissues in F1 and F2 pups that were euthanised at PND 21 without active dosing.
In the F1 cohort 2A and 2B animals used for neurological investigations (euthanised on PND 21 or dosed from PND 21 until at least PND 78-80), there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions.
There were no test item-related gross findings in the entire study, and there were only 6 premature deaths, all of which were considered not test item-related.
There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility.
In conclusion, administration of the test item by once daily (or twice daily 3 hours apart on some occasions) oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no test item-related deaths. There were occasional observations of decreased activity, abnormal, uncoordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose level-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body weight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirmed by overall ambulation values, or any associated clinical findings, detailed functional observations or brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 males only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels (up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertrophy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney weights were seen in both sexes. In neurological investigations, there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions. There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1 adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the in-life and pathological findings were considered not to be adverse. For F1 and F2 developmental parameters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on the pup numbers, survival or development.
- Endpoint:
- extended one-generation reproductive toxicity - with F2 generation and developmental neurotoxicity (Cohorts 1A, 1B with extension, 2A and 2B)
- Type of information:
- experimental study
- Remarks:
- Performed on source substance.
- Adequacy of study:
- key study
- Study period:
- April 2021 - April 2022
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Reason / purpose for cross-reference:
- read-across source
- Remarks:
- Hydroxycyclohexyl phenyl ketone
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 424. Neurotoxicity Study in Rodents
- Version / remarks:
- 21 July 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 443 (Extended One-Generation Reproductive Toxicity Study)
- Version / remarks:
- 25 June 2018
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Justification for study design:
- At this time, studies in laboratory animals provide the best available basis for extrapolation to humans and are required to support regulatory submissions. Acceptable models that do not use live animals currently do not exist. The Han Wistar rat was chosen as the animal model for this study as it is an accepted rodent species for nonclinical toxicity testing by regulatory agencies. The total number of animals to be used in this study is considered to be the minimum required to properly characterise the effects of the test item. This study has been designed such that it does not require an unnecessary number of animals to accomplish its objectives.
The oral route of exposure was selected because this is the intended route of human exposure. In the OECD 414 study (prenatal developmental toxicity), female rats dosed at 900 mg/kg bw/day had treatment related clinical signs like piloerection and lethargy, among others, that occurred over several days but mostly resolved by the end of treatment. Females had lower body weights and weight gains from Days 15-20 post coitum and absolute and relative food consumption was also lower Days 6-17 post coitum. There were no unscheduled deaths during the course of the study and no maternal toxicity was observed in the 100 or 300 mg/kg bw/day groups. No developmental toxicity was observed in the 100, 300 and 900 mg/kg bw/day groups. Based on the results in this prenatal developmental toxicity study (OECD 414, conducted with GLP certification) the maternal No Observed Adverse Effect Level (NOAEL) for the test substance was established as being 300 mg/kg bw/day and developmental NOAEL was established as 900 mg/kg bw/day.
The proposed dose levels for this study were: Low dose 100 mg/kg bw/day, Mid dose 300 mg/kg bw/day, High dose 900 mg/kg bw/day on the basis that evidence of systemic toxicity will be observed at the top dose level. - Specific details on test material used for the study:
- - Identification: Omnirad 184
- Alternate identification: Hydroxycyclohexyl phenyl ketone
- EC number: 213-426-9
- CAS number: 947-19-3
- Physical description: Light white solid
- Storage condition of test material: Ambient, protected from light
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the
exposure medium) and during storage: Test Item is stable in the vehicle when prepared and stored
under the same conditions at concentrations bracketing those used in the present study - Species:
- rat
- Strain:
- Wistar
- Remarks:
- Han Wistar (Crl:WI(Han))
- Details on species / strain selection:
- The Han Wistar rat was chosen as the animal model for this study as it is an accepted rodent species for nonclinical toxicity testing by regulatory agencies, it is also the preferred model for this OECD guideline study.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River UK, Margate, Kent, UK
- Age at study initiation: ca 6-7 weeks for males and 5-6 weeks for females
- Weight at study initiation: 150 to 250 g (males)/100 to 150 g (females)
- Housing: Group housed (up to 3 or 4 animals of the same sex and same dosing group together). A
few days prior to mating, F0 and Cohort 1B males were be transferred to individual cages with solid
bottoms. F0 and Cohort 1B females were transferred to these cages for mating. Mated F0 and Co
hort 1B females were transferred to individual solid bottomed cages. White paper tissue was supplied
as nesting material from Gestation Day 20. F0 and Cohort 1B females with litters were retained in t
his type of cage until termination. On a suitable day after completion of mating, the F0 and Cohort 1B
males were re-housed with their original cage mates.
- Diet (e.g. ad libitum): Special Diet Services VRF1. Ad libitum, except during designated procedures
- Water (e.g. ad libitum): Public supply tap water. Freely available to each animal from water bottles
(except during
designated procedures).
- Acclimation period: The F0 animals were allowed to acclimate to the Test Facility rodent toxicology
accommodation for 10 days before the commencement of dosing (replacement animals at least 5
days).
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23°C
- Humidity (%): 40 - 70%
- Air changes (per hr): Ten or more air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark (except during designated pro
cedures)
IN-LIFE DATES: From: 12 Apr 2021 To: 21 Dec 2021 - Route of administration:
- oral: gavage
- Vehicle:
- CMC (carboxymethyl cellulose)
- Remarks:
- CMC (carboxymethyl cellulose) 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q Water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: Dose formulations divided into aliquots where required to
allow them to be dispensed on each dosing occasion
VEHICLE
- Justification for use and choice of vehicle: Carboxymethylcellulose, analysis performed in a s
eparate study showed good stability in this vehicle
- Concentration in vehicle: 10, 30, 90mg/mL
- Amount of vehicle: 10ml/kg - Details on mating procedure:
- Frequency: F0 - Day 71
Cohort 1B – PND 107-111
Procedure: Pairing was on a 1 male to 1 female basis. During the evening (after 5 pm), females
were housed with their allocated co-group male partner. The animals were paired in ascending numerical order. Vaginal lavages were taken early each morning from the day of pairing until mating
had occurred and the stage of estrous observed in each vaginal lavage was recorded. The day of
detection of a copulatory plug in situ and/or of sperm in the lavage was designated Gestation Day 0.
The pairing period for each pair of animals was a maximum of 14 nights (unless a longer period was
deemed appropriate).
If evidence of mating was not observed by the end of the pairing period, the female was separated from the male during the morning following the last night of pairing and treated as if mating had occurred during that night. Procedures for that female were continued as if it had mated on the last
night of pairing.
For each female the time taken to show a positive mating sign and the number of failed opportunities t
o mate (estruses passed without a sign of mating) was evaluated. - Analytical verification of doses or concentrations:
- yes
- Remarks:
- Concentration & Homogeneity, and stability analysis
- Details on analytical verification of doses or concentrations:
- Analyses was performed by Gas Chromatography (GC) using a validated analytical procedure.
Concentration and Homogeneity Analysis
Sample Allocation: 2 for analysis, 3 for backup for control; 2 for analysis and 3 for backup per stratum
for Groups 2-4.
Sampling Containers: Appropriate sized volumetric flasks
Storage Conditions: Temperature set to maintain 4°C, protected from light
Acceptance Criteria: For concentration: mean sample concentration results within or equal to
± 15% of theoretical concentration. Each individual sample concentration result within or equal to ±
20%. For homogeneity, relative standard deviation (RSD) of concentrations of ≤ 10% for each group.
Stability analyses performed previously in conjunction with 426730 demonstrated that the Test Item
is stable in the vehicle when prepared and stored under the same conditions at concentrations
bracketing those used in the present study. - Duration of treatment / exposure:
- F0 Males: 10 weeks prior to mating and continuing throughout and after mating until the day before t
ermination.
F0 Females: 10 weeks prior to mating and continuing throughout mating and gestation until at least
LD 21.
Cohort 1A: From PND 21 until the day before necropsy
Surplus Animals: From PND 21 until the day before necropsy
Cohort 1B: From PND 21 until the day before necropsy
Cohort 2A: From PND 21 until the day before necropsy - Frequency of treatment:
- Once daily.
From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day. - Details on study schedule:
- F0
Animal Arrival (F0): 02 Apr 2021
Replacement Animals: 09 Apr 2021
Initiation of Dosing: 12 Apr 2021
F0 Start of Estrous Assessment: 07 Jun 2021
F0 Pairing for Mating: 21 Jun 2021
F0 Parturition (Expected): from ca 13 Jul 2021
F0 Dams (Lactation Day (LD) 21) and Unselected F1 Pups Necropsies: from ca 03 Aug 2021
F0 Male Necropsies (Including Computer Assisted Sperm Assessment (CASA)): from 26 Aug 2021
F1
First expected Post Natal Day (PND) 1: from 14 Jul 2021
PND 4 Culled Pups Necropsy and Terminal Blood Sampling: from 17 Jul 2021
Initiation of Dosing (Cohorts 1A + surplus animals, 1B and 2A): from ca 04 Aug 2021
PND 21 Unselected Pups Necropsy and Terminal Blood Sampling: from ca 04 Aug 2021
Cohort 2B Necropsies (PND 21): from 04 Aug 2021
Cohort 2A Necropsies (PND 78-80): from 29 Sep 2021
Cohort 1A Necropsies (PND 91) (males include CASA): from 12 Oct 2021
Surplus animals Necropsies (ca PND 52): 06 Sep 2021
F2
Cohort 1B - Start of Estrous Assessment: 19 Oct 2021
Cohort 1B - Mating: from 02 Nov 2021
Cohort 1B – Parturition (Expected): from 24 Nov 2021
Cohort 1B PND4 Culled Pups Necropsy and Terminal Blood Sampling: from 28 Nov 2021
Cohort 1B – Dams (LD 21) And F2 litter Necropsies: from 15 Dec 2021
Cohort 1B Male Necropsies: from 09 Dec 2021
Completion of In-life: 21 Dec 2021 (Last date of necropsy) - Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- Control
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Dose / conc.:
- 900 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 10 animals per sex per dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale:
- F0 animals were randomly assigned to groups. Males and females were randomised separately.
- Fasting period before blood sampling for clinical biochemistry: animals not fasted - Positive control:
- n/a
- Parental animals: Observations and examinations:
- F0 Animals
CAGE SIDE OBSERVATIONS:
Pre and Postdose Observations, All F0 animals.
Frequency: Once predose and up to 4 hours postdose after the end of each group for each sex.
DETAILED CLINICAL OBSERVATIONS:
All F0 animals Weekly; from at least Week 1 and throughout the study.
BODY WEIGHT:
All F0 animals.
Males: weekly beginning Week -1.
Females: weekly beginning Week -1 until pairing for mating and then on GD 0, 7, 14 and 20 and LD 1,
4, 7, 14 and 21.
FOOD CONSUMPTION:
All F0 animals.
Males: weekly from Week -1 until pairing for mating. Then weekly after mating and re-housing.
Females: weekly beginning week -1 until pairing for mating and then on GD 0-7, 7-14 and 14-20, and
LD 1-7, 7-14 and 14-21
WATER CONSUMPTION:
All F0 animals. Regular basis throughout the study.
Mortality:
All F0 animals. At least twice daily (once at the start and once towards the end of the working day)
beginning upon arrival through termination/release. - Oestrous cyclicity (parental animals):
- Estrous Cycle Monitoring (Number of Estrous Cycles and Mean Cycle Length):
All F0 females. From 2 weeks prior to pairing until day of detection of a copulatory plug in situ and/or of sperm in the lavage. Until either mating was detected or the end of the mating period. Also, on the morning of necropsy. - Sperm parameters (parental animals):
- Parameters examined in F0 and Cohort 1A Males:
Computer Aided Sperm Assessment (CASA)
From all F0 and Cohort 1A males at terminal euthanasia, the right cauda epididymis was placed in 0.3% BSA in Medium 199 (as per SOP/PAT/069) and the sperm was allowed to “swim out” into the medium. An appropriate dilution of the sperm suspension was prepared and examined using a Ham ilton Thorne sperm motility analyser.
Sperm Count and Morphological Analysis
The cauda epididymis was minced and suspended. Dilutions of this sperm suspension were counted using a haemocytometer to obtain a total sperm count which was expressed per cauda epididymis and per gram of cauda epididymis. Optionally, the cauda epididymis was frozen prior to assess ment. From all samples of the sperm suspension described in the preceding paragraph, a sperm smear was prepared and stained with eosin Y solution. At least two hundred sperm per animal were evaluated for morphological abnormalities
Spermatid Count
The right testes was decapsulated and homogenised. The number of homogenisation resistant spermatids in dilutions of this suspension were counted using a haemocytometer to obtain a total spermatid count which was expressed per testis and per gram of testis. Optionally, the testis was frozen prior to assessment.
F0 Animals and Cohort 1A
Testis were weighed. - Litter observations:
- STANDARDISATION OF LITTERS
For females, on Lactation Day 4, litter size was standardised to 8 pups per litter (and if possible, co
mprised of 4 males and 4 females). Any additional pups were selected at random for use in bloodsampling
or necropsy procedures. Where 4 males and 4 females were not available on PND 4, addi
tional pups of the opposite sex were used to ensure there were a total of 8 pups. Where the total nu
mber of pups in the litter on PND 4 was 8 or fewer, no adjustment was performed. Where practicable,
any pups that were found dead or were killed during the postnatal period were sexed and appropriatel
y examined as above. Any externally abnormal decedent pup was preserved in 10% neutral buffered
formalin; externally normal pups were discarded.
General In-life Assessments – F1 Animals (Cohorts 1A + Surplus Animals, 1B and 2A)
- Mortality:
All F1 animals At least twice daily (once at the start and once towards the end of the working day)
beginning upon arrival through termination/release.
- Pre and Postdose Observations:
All F1 animals. Once predose and up to 4 hours postdose after the end of each group for each sex.
- Detailed Clinical Observations:
All F1 animals. Weekly from weaning, starting on a suitable day within one week of weaning of the
majority of litters (weaning on PND 21).
- Individual Body Weights:
All F1 animals. On day of weaning and then weekly, starting on a suitable day within one week of wea
ning of the majority of litters (beginning of nominal Week 4 of age). For Cohort 1B females only, we
ekly body weights were continued until pairing for mating and then mated females were weighed on
Gestation Day 0, 7, 14 and 20 and Lactation Day 1, 4, 7, 14 and 21. A body weight may also have
been taken on Lactation Day 0 for dosing purposes only. Any Lactation Day 0 body
weights were not reported but were maintained in the study data. Pups were weighed individually on
PND 1, 4, 7, 14 and 21.
- Food Consumption:
All F1 animals. Excluding surplus animals. Weekly, starting on a suitable day within one week of wea
ning of the majority of litters (Nominal Week 4 of age).
For Cohort 1B animals, food consumption was recorded until pairing for mating.
1B Mated females: over the periods Gestation Days 0-7, 7-14 and 14-20, and Lactation Days 1-7,
7-14 and 14-21.
1B Males: weekly after mating and re-housing.
- Estrous Cycle Monitoring:
F1 females. Cohort 1A + Surplus Animals. From the day after vaginal patency, continuing until one
estrous smear had been identified.
F1 females. Cohort 1A. Smears were also taken for at least 14 consecutive days prior to necropsy, up
to and including the day of necropsy.
F1 females. Cohort 1B. From 2 weeks prior to pairing until day of detection of a copulatory plug in situ
and/or of sperm in the lavage.
- Water Consumption:
All F1 animals. Regular basis throughout the study.
PARAMETERS EXAMINED
The numbers of live and dead pups born in each litter were recorded as soon as possible after
completion of parturition on LD 0. The live pups were counted and examined daily for the presence o
f milk in the stomach and for any externally visible abnormalities up to and including PND 4 and then on PND 7, 14 and 21. Individual pup weights were taken for each litter on PND 1, 4, 7, 14 and 21. A
head count of the pups was performed each day.
Pre-Weaning Physical Development of F1 and F2 Pups:
Ano-genital distance was measured for the pups of both sexes on PND 1. Nipple retention was
assessed in males on PND 13.
Assessment of Sexual Maturation Cohorts 1A+ Surplus Animals, 1B and 2A:
Commencing on PND 28, females were examined daily for vaginal opening. The day on which the va
gina became open was recorded, as was the body weight on that day.
Commencing on PND 35, males were examined daily for balanopreputial separation. The day on wh
ich separation occurred was recorded, as was the body weight on that day.
GROSS EXAMINATION OF DEAD PUPS:
Pups Found Dead or Euthanised Prematurely Before PND 14: Where practicable, these animals were
sexed and then checked for the presence of milk in the stomach and for the presence of any extern
ally visible abnormalities. Any externally abnormal pups were fixed in 10% neutral buffered formalin
for optional further examination. Externally normal pups were discarded.
Pups Found Dead or Euthanised Prematurely On or After PND 14: These animals were subject to a
gross necropsy. An external examination was followed by an inspection of the cranial, thoracic and
abdominal contents. Internal sex was also confirmed. Representative samples of any abnormal t
issues were taken and fixed in neutral buffered 10% neutral buffered formalin. These carcasses were
then discarded.
ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:
Perfusion fixation, neuropathology and brain morphometry was performed for all animals. The fixed b
rains were removed and weighed, and the length and maximum width of the brain was measured for
all animals selected for neuropathology.
Brain, dorsal root ganglion, and spinal cords tissues were prepared for neuropathology examination.
Neuropathology and brain morphometry were performed.
ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:
Lymph nodes were weighed to evaluate immunotoxic effects from 10 rats/sex/group from Groups 1-4.
One-half of the spleen was used for immunophenotyping assessment from 10 rats/sex/group from
Groups 1-4. - Postmortem examinations (parental animals):
- GROSS NECROPSY
- Gross necropsy consisted of a complete necropsy examination, which included evaluation of the
carcass and musculoskeletal system; all external surfaces and orifices; cranial cavity and external
surfaces of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and
tissues. The reproductive tracts of all F0 females were be examined for signs of implantation, the
number of any implantation sites being recorded. The total number of corpora lutea
graviditatis were recorded for each female. For F0 females necropsied on GD 24 as they had failed to
produce a litter, the uteri of all non-pregnant females were examined for implantation sites and fixed
in 10% neutral buffered formalin. On GD 24, if a female was found to be pregnant, the number and
type of any implantation sites were recorded and the total number of corpora lutea graviditatis were
recorded also.
HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination and weighed, respectively: Artery,
aorta, Body cavity, nasal (Processing at the level of the first
upper molar teeth, to include olfactory bulbs and nasopharynx and/or nasopharyngeal duct), Bone
marrow sternum, Bone marrow smear, bone sternum, brain, epididymis, esophagus, eye, dorsal root
lumbar ganglion, adrenal gland, clitoral gland, lacrimal gland, harderian gland, mammary gland, para
thyroid gland, pituitary gland, preputial gland, prostate gland, submandibular gland, sublingual gland,
parotid gland, seminal vesicle gland, thyroid gland, zymbal's gland, Gut-associated lymphoid tissue, heart, kidney, cecum, colon, rectum, larynx, liver, lung, axillary lymph node, mandibular lymph node,
mesenteric lymph node, gastrocnemius muscle, quadriceps muscle, optic nerve, sciatic nerve, tibial
nerve, ovary, oviduct, pancreas, skin, duodenum ileum, jejunum, spinal cord, spleen, stomach, testis,
thymus, tongue, trachea, ureter, urinary bladder, uterus/cervix, vagina, vas deferens.
Immunophenotyping Sample Collection, Processing, and Analysis - F0 Animals and Cohort 1A:
Spleen samples (approximately half of the spleen) were taken from 10 rats/sex/group of the F0 and
Cohort 1A animals at necropsy and will be collected into tubes containing RPMI-1640 medium and
placed immediately on wet ice until processing. The remainder of the spleen was preserved in 10%
neutral buffered formalin.
Upon receipt at the flow cytometry laboratory, the samples were stored on wet ice and were analysed
as follows:
The cellular antigens and cell populations identified below were quantified, using specific antibodies
against the marker antigens. The samples were analysed using the
Test Facility validated analytical method 996094:
Antigen markers:
CD45+CD3-CD45R+ (B cells)
CD45+CD3+CD45R- (Total T cells)
CD45+CD3+CD45R-CD4+CD8- (T helper cells)
CD45+CD3+CD45R-CD4+CD8- (Cytotoxic T cells)
CD45+CD3-CD45R-CD161+ (Natural killer cells)
Parameters to be Reported - % of lymphocytes - Postmortem examinations (offspring):
- Necropsy (F1 and F2 Culled PND 4 and Unselected PND 21 pups):
Culled pups on PND 4 were necropsied. Necropsy consisted of external examination followed by
macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities
in situ. Samples of any grossly abnormal tissues were preserved in 10% neutral buffered formalin or
other appropriate fixative. The carcasses were discarded.
All unselected pups on PND 21 were necropsied. Necropsy consisted of external examination
followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdomina
l cavities in situ. Representative samples of abnormal tissues were preserved in 10% neutral buffere
d formalin or other appropriate fixative. For F1 and F2 pups, tissue samples and weights of the foll
owing tissues were taken from 10 male and 10 female pups per group and were preserved in 10%
neutral buffered formalin or other appropriate fixative: Brain, mammary gland, parathyroid gland,
thyroid gland, liver, ovary, spleen, testes, thymus.
Where 10 pups of each sex were not available for organ weight and tissue collection, additional pups
of the opposite sex were selected such that 20 animals per group had organ weights and tissues
collected.
Necropsy (Cohort 1B Animals):
Animals were subjected to a complete necropsy examination, which included evaluation of the carcas
s and musculoskeletal system; all external surfaces and orifices; cranial cavity and external surfaces
of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and tissues.
The reproductive tracts of all F1 females were examined for signs of implantation, the number of any
implantation sites being recorded. The total number of corpora lutea graviditatis were recorded for
each female. For F1 females necropsied on GD 24 as they had failed to produce a litter, the uteri
of all non-pregnant females were fixed in 10% neutral buffered formalin after being examined for
implantation sites. On GD 24, if a female was found to be pregnant, the number and type of any impl
antation sites were recorded and the total number of corpora lutea graviditatis were recorded also.
Transcardial Perfusion Fixation of Cohorts 2A and 2B:
All animals were subjected to a limited examination, with special attention being paid to the reproducti
ve organs.
The head (with brain exposed), spinal cord and hindlimbs (Cohort 2A) and head (with brain exposed)
(Cohort 2B) was placed in 10% neutral buffered formalin and allowed to fix for at least 7 days. The
fixed brains were removed and weighed, and the length and maximum width of the brain was measured for all animals selected for neuropathology. Subsequently, the brain was fixed in 10% neutral buffered formalin together with selected PNS tissues. - Statistics:
- Body Weight Changes: Calculated between each scheduled interval
Food Consumption: Calculated between appropriate intervals
Organ Weight Relative to Body Weight: Calculated against the terminal body weight for scheduled
intervals
Organ Weight Relative to Brain Weight: Calculated against the brain weight for scheduled intervals
Descriptive Statistical Analyses:
Means, standard deviations, percentages, numbers, and/or incidences have been reported as appropriate by dataset.
Inferential Statistical Methods:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and have been reported at the 1% and 5% levels.
The pairwise comparisons of interest are listed below:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1
Analyses were performed according to the matrix below when possible but excluded any group with less than 3 observations.
See Table 43 below.
Parametric/Non-parametric:
Levene’s test was used to assess the homogeneity of group variances.
The groups were compared using an overall one-way ANOVA F-test if Levene’s test was not significant or the Kruskal-Wallis test if it was significant. If the overall F-test or Kruskal Wallis test was found to be significant, then pairwise comparisons were conducted using Dunnett’s or Dunn’s test, respectively.
Non-Parametric:
The groups were compared using an overall Kruskal-Wallis test. If the overall Kruskal-Wallis test was found to be significant, then the above pairwise comparisons were conducted using Dunn’s test (equivalent to Wilcoxon Rank-Sum test in Nevis 2012 tables). - Reproductive indices:
- Female Mating Index = Number of Females with Evidence of Mating (or no confirmed mating date and pregnant) / Number of Females Paired
Female Fertility Index = Number of Pregnant Females / Number of Females with Evidence of Mating (or no confirmed mating date and pregnant)
Female Pregnancy Index = Number of Pregnant Females / Number of Females Paired
Male Mating Index = Number of Males with Evidence of Mating (or female partner confirmed pregna
nt) / Number of Males Paired
Male Fertility Index = Number of Males Impregnating a Female / Number of Males with Evidence of
Mating (or female partner confirmed pregnant)
Male Pregnancy Index = Number of Males Impregnating a Female / Number of Males Paired
The following natural delivery/reproductive parameters were included, as appropriate:
Gestation Length: The gestation length was calculated from GD 0 to the day the first pup was observed.
Gestation Index:
Percentage of pregnancies that result in birth of live litters = (Number of Animals with Live Offspring / Number of Animals Pregnant) x 100 - Offspring viability indices:
- Live Birth Index:
Percentage of pups born alive = (Number of Live Newborn Pups / Number of Newborn
Pups) x100
Sex Ratio (% males):
Percentage of male pups per litter = (Number of Live Male Pups / Total Number of Live Pups) x100
Viability Index:
Percentage of pups born that survive 4 days postpartum = (Number of Live Pups on Day 4 Postpartum / Number of Live Newborn Pups) x100
Lactation Index:
Percentage of pups that survive 21 days postpartum = (Number of Live Pups on Day 21 Postpartum / Number of Live Pups on Day 4 (postculling) Postpartum) x100
Post-Implantation Loss/Litter = (Number of Implants – Total Newborn Pups / Number of Implants) x100 - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- At 900 mg/kg/day, decreased activity was observed for one male on Day 2 and abnormal, unco
ordinated gait for 3 males and 4 females between Days 2 to 4. Decreased activity, abnormal, uncoordinated gait or low carriage were occasionally noted for a further 8 females between Days 5 to 70 and decreased activity on one occasion for each of 2 females during lactation. These findings were considered to be test item-related.
Salivation and/or ploughing (also a chewing action for one female at 900 mg/kg/day) were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, on multiple days throughout the dosing period. The incidence of these findings was dose level-related.
Animal 4515F (900 mg/kg/day) had abnormal clincal observations of irregular respiratory rate, hunched posture, erect fur, cold to touch, prominent backbone, abnormal gait, decreased activity and red discharge from vagina the day after parturition (LD 1) but these mostly resolved before scheduled termination. The clinical observations were considered to be likely due to the trauma of giving birth rather than being test item-related.
Animal 4525F (900 mg/kg/day) had abnormal clincal observations of irregular/decreased respiratory rate, erect fur, eyes partly closed, decreased activity up to 2-4 hours postdose on LD 7 only. No
similar findings were observed for this animal during the dosing period and as a cause could not be
determined, these findings were likely related to the dosing procedure. - Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- None of the unscheduled deaths were considered to be test item-related.
On Study Day 40, approximately 3 hours after dosing, Animal 4510F given 900 mg/kg/day was found
with clinical signs of irregular respiratory rate, hunched posture, partly closed eyes, erect fur, unc
oordinated and abnormal gait and decreased activity and was euthanised. There were no findings
at necropsy or microscopically to suggest the likely cause of death. This animal had not had any
abnormal clinical signs before this (apart from salivation and ploughing) and had consistently gained
body weight.
Animals 2512F and 4502F given 100 or 900 mg/kg/day respectively, were euthanised on LD 1 or LD
0 and due to the very young age of the pups, they were also euthanised. Animal 2512F had clinical
signs of irregular respiratory rate, hunched posture, pale skin and cold to touch, erect fur, uncoor
dinated and abnormal gait in the afternoon of LD 1. Animal 4502F was found subdued, cold to touch,
prostrate/lying on side with decreased respiratory rate approximately 1 hour after dosing on LD 0. Th
ere were no findings at necropsy or microscopically to suggest the likely cause of death and both an
imals had not had any abnormal clinical signs before this (apart from salivation and ploughing for 4
502F) and had consistently gained body weight. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See Tables 1-4.
For males given 900 mg/kg/day, there was slightly lower body weight gain over the dosing period resulting in male body weight being 3.6% lower than the concurrent control mean value by Day 134. For females given ≥ 100 mg/kg/day, a dose level-related higher body weight gain of up to 10.3% at 900 mg/kg/day compared to controls was noted during the pre-pairing period and this slightly higher body weight was generally maintained throughout gestation and lactation.
There was no test item-related effect on male body weight at 100 or 300 mg/kg/day. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- There were occasional slightly higher food consumption values of up to 14.4% of controls noted for
males at 300 or 900 mg/kg/day up to Day 106. At ≥100 mg/kg/day for males from Days 106 to 134,
there was higher food consumption of up to 17.5%. For females given ≥100 mg/kg/day, there was an
overall dose level-related higher food consumption of up to 20.6% of control values during the prepairing
period, and during gestation and lactation, food consumption was higher for females by up to
13.2% at 300 or 900 mg/kg/day only. - Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Haematology
See tables 18-19.
Coagulation
See tables 15-16.
All differences in haematology or coagulation parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control values and/or were of a magnitude of variation commonly observed in rats. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 12-13.
At ≥300 mg/kg/day for males and ≥100 mg/kg/day for females, triglycerides were lower than controls by up to 0.75-fold, and for males only at ≥300 mg/kg/day, cholesterol higher by up to 1.30-fold. - Endocrine findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of
necropsy in F0 adult males given 300 or 900 mg/kg/day.
There were no test item-related effects for F0 adult males given 100 mg/kg/day or F0 adult females gi
ven up to 900 mg/kg/day or for culled pups on PND 4 or unselected pups on PND 21.
There were no test item-related changes to T4 levels at up to 900 mg/kg/day for samples taken on the
morning of necropsy for the F0 generation animals, for culled pups on PND 4 or unselected pups on
PND 21.
The differences in these parameters were considered not to be test item-related based on their small
magnitude, inconsistent direction, absence of a dose response, general overlap of individual values
with the range of control and/or were of a magnitude of variation commonly observed in rats. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See tables 27-28.
All differences in urine parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats. - Immunological findings:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Test item-related Microscopic Findings
In the liver, there was diffuse centrilobular hepatocyte hypertrophy of minimal to mild severity. For
males only, this was seen in all dosage groups, whereas for females this was only identified in ani
mals dosed at 900 mg/kg/day. Although this was also found in one control animal (M1021), the
incidence was higher in animals treated with Omnirad 184 and followed a dose-related distribution.
This finding was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in f
emales dosed at 900 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184, were correlated to the histopatholo
gic findings of centrilobular hypertrophy. This correlation was evident in the males dosed at ≥ 100 mg/
kg/day and females dosed at 900 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal
severity. This was characterised by an increase in the number of goblet cells. For males only, this was
seen in at ≥ 300 mg/kg/day, whereas for females this was only identified in animals dosed at 900 mg/
kg/day. Although this was also found in one control animal (M1013), the incidence was convincingly
higher in animals treated with Omnirad 184 at ≥ 300 mg/kg/day and showed a dose-related distribut
ion. This finding was therefore considered test item-related in males dosed at ≥ 300 mg/kg/day and in
females dosed at 900 mg/kg/day.
Additional Microscopic Findings:
In the spleen, there was a slightly higher incidence of increased hematopoietic cells with a minimal
severity, primarily in males given Omnirad 184 at a doses of 100 mg/kg/day and 900 mg/kg/day. A
slightly higher incidence was seen in females dosed at 900 mg/kg/day only. However, due to the
overall low incidences and because this is a common background finding, it was considered not test i
tem-related.
Other microscopic findings were of the nature commonly observed in this strain and age of rat, or
occurred at a similar incidence in control and treated animals, and, therefore, were considered not to
be test item-related. - Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- There was no effect on the number or length of estrous cycles during the 2 weeks before pairing or during the mating period at up to 900 mg/kg/day, compared with controls.
On the morning of necropsy, while the majority of females in all groups were still in diestrus, a few in each group had come back into estrus. - Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- There were no test item-related effects on the percentage of motile sperm, progressively motile sperm or straight line velocity at up to 900 mg/kg/day.
F0 control male 1020M control was paired with 1520F which was found not to be pregnant at scheduled necropsy. The sperm parameters for male 1020M were within the normal ranges.
F0 Animal 2011M given 100 mg/kg/day had a very low sperm motility of 2% at necropsy, however, the female it had been paired with 2511F, became pregnant and littered successfully.
At 900 mg/kg/day, F0 Control 1024M was paired with 1524F, and 4010M was paired with 4511F. These females failed to produce litters, however, the sperm parameters for the males were within the normal ranges.
Sperm Count and Morphological Analysis:
There were no test item-related effects on sperm count or morphology at up to 900 mg/kg/day.
Spermatid Count:
There was no test item-related effect on the number of spermatids at up to 900 mg/kg/day. - Reproductive performance:
- no effects observed
- Description (incidence and severity):
- There were no test item-related effects on male or female mating performance or fertility, or duration of gestation in females, at up to 900 mg/kg/day.
F0 control 1520F was found not to be pregnant at scheduled necropsy.
F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early resorption) and 4511F was not pregnant.
F0 Animals 1501F, 1502F, 1506F, 1518F, 1521F (control), 2517F (100 mg/kg/day), 350F2 (300 mg/kg/day) and 4523F (900 mg/kg/day) did not have a positive mating sign during pairing and therefore gave birth before their expected GD 21.
See table 20, table 23, and table 25. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- body weight and weight gain
- food consumption and compound intake
- clinical biochemistry
- organ weights and organ / body weight ratios
- histopathology: non-neoplastic
- reproductive function (oestrous cycle)
- reproductive function (sperm measures)
- reproductive performance
- Critical effects observed:
- no
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- Salivation and/or ploughing were noted for all animals (except Surplus Animal 3650F) given ≥300 mg/kg/day and for a few animals given 100 mg/kg/day, on several days throughout the dosing period. The incidence of these findings was dose level-related.
Surplus Animal 4146M had erect fur on Day 5 postdose and 4147M had irregular respiratory rate, erect fur, eyes partly closed and abnormal gait on Day 4 postdose. Both animals (given 900 mg/kg/day) did not have any similar findings during the dosing period and as a cause could not be determined, these abnormal findings were likely related to the dosing procedure.
For all generations, the other clinical observations recorded were considered not to be test item-related as they were of low incidence, are commonly seen during gestation or lactation, were also seen in some control animals or were not dose level-related. - Mortality / viability:
- mortality observed, non-treatment-related
- Description (incidence and severity):
- Animal 3608F given 300 mg/kg/day had convulsions and died within minutes of receiving its first dose
on Day 1. Grossly, the lungs had pale discoloration and abnormal consistency and the trachea had fr
othy pale content. Microscopically, there was multifocal mixed cell infiltration of the alveolar space of
the lung which was considered to correlate with the gross findings in the lung. Overall, these findings
were considered to have been related to the dosing procedure and were therefore considered to be
the cause of death.
Control Animal 1628F was euthanised on LD 1 because all pups in the litter died. At necropsy, a
pale pancreas was noted but no microscopic findings could be correlated and no cause of death w
as identified. The animal had not had any abnormal clinical signs before this and had consistently g
ained body weight.
Animal 4634F was found dead in the afternoon of LD 18. At necropsy, in the abdominal cavity, there
was dark red fluid accumulation and a soft red mass adherent to the stomach. Microscopically this
was correlated with a well demarcated hematocyst of marked severity in relation to hepatic tissue. Thi
s finding was considered to be the animal’s cause of death. As it’s pups had not been weaned, they w
ere also euthanised. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 5-11.
For animals in Cohorts 1A and 1B, there was a general trend of slightly lower body weight gain (up to
7.0%) for males at 900 mg/kg/day compared to controls, but no test item related-effects for males at 100 or 300 mg/kg/day or females at ≥100 mg/kg/day during pre-pairing, gestation or lactation.
In Cohort 2A and for Surplus animals, body weight gain was variable and did not follow any trend. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- There were occasional higher food consumption values of up to 19.6% of controls noted for males
at 300 or 900 mg/kg/day, in some F1 generation cohorts. There was a general trend of higher food
consumption for females of up to 22.4% higher at 300 or 900 mg/kg/day but during gestation and la
ctation female food consumption was similar to controls. - Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See table 20.
All differences in haematology or coagulation parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control values and/or were of a magnitude of variation commonly observed in rats. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- See table 14.
At ≥100 mg/kg/day for males and females, triglycerides were lower than controls by up to 0.62-fold,
and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold.
For both generations, all other differences in clinical chemistry parameters were considered not to be test item-related based on their small magnitude, inconsistent direction, absence of a dose response, general overlap of individual values with the range of control and/or were of a magnitude of variation commonly observed in rats. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See table 29 .
All differences in urine parameters were considered not to be test item-related
based on their small magnitude, inconsistent direction, absence of a dose response, general overla
p of individual values with the range of control and/or were of a magnitude of variation commonly ob
served in rats. - Sexual maturation:
- no effects observed
- Description (incidence and severity):
- There was considered to be no effect on sexual maturation for F1 males or females at up to 900 mg/kg/day.
- Anogenital distance (AGD):
- no effects observed
- Description (incidence and severity):
- There was no effect on anogenital distance or normalised anogenital distance for males or females at up to 900 mg/kg/day.
- Nipple retention in male pups:
- no effects observed
- Description (incidence and severity):
- On PND 13, there was no effect on nipple retention in male pups i.e. no nipples were present, at dose levels up to 900 mg/kg/day.
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- F1 Cohort 1A
Test item-related Organ Weight Differences:
Liver weights in animals dosed at ≥ 300 mg/kg/day were statistically significantly higher than the controls and showed a dose-related response. In males, liver weights were up to ~24% higher and in females up to ~29% higher (relative to body weight at 900 mg/kg/day). This was therefore considered test item-related.
Kidney weights in animals dosed at ≥ 300 mg/kg/day in males and at ≥ 100 mg/kg/day in females, were statistically significantly higher than the controls and showed a dose related response. In males, kidney weights were up to ~23% higher and in females up to ~16% higher (relative to body weight at 900 mg/kg/day). Although this was not correlated to any microscopic findings, this was considered test item-related in view of the similar weight differences recorded in the F0 and F1 cohort 1B animals.
Additional Organ Weight Differences:
Testis weights relative to body weight were statistically significantly higher by ~11% only at 900 mg/kg/day compared to the controls. However, males dosed at 900 mg/kg/day had a lower terminal body weight and in view of the minor weight difference and no microscopic findings that could be correlated. This was therefore considered incidental.
Ovary weights were statistically significantly higher by ~19% (relative to body weight at 300 mg/kg/day). However, no histologic findings could be correlated to this finding and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Thyroid/parathyroid gland weights were statistically significantly higher by ~21% (relative to body weight) in female animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, thyroid hormone was not increased in a noteworthy fashion, and no similar differences were found in any other generation or cohort of animals, so this was considered incidental.
Seminal vesicle weights were statistically significantly higher by ~22% (relative to body weight) in animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be correlated to this finding, so this was considered not test item-related.
There were individual organ weight values that were different from their respective controls. There were, however, no patterns or correlating data to suggest these values were test item-related.
F1 Cohort 1B
Test item-related Organ Weight Differences:
Liver weights in animals dosed at ≥ 300 mg/kg/day were statistically significantly higher than the co
ntrols and showed a dose-related response. In males, liver weights were up to ~24% higher and in
females up to ~29% higher (relative to body weight at 900 mg/kg/day). This was therefore considered
test item-related.
Kidney weights in animals dosed at ≥ 300 mg/kg/day in males and at ≥ 100 mg/kg/day in females,
were statistically significantly higher than the controls and showed a dose related response. In males,
kidney weights were up to ~23% higher and in females up to ~16% higher (relative to body weight
at 900 mg/kg/day). Although this was not correlated to any microscopic findings, this was considere
d test item-related in view of the similar weight differences recorded in the F0 and F1 cohort 1B an
imals.
Additional Organ Weight Differences:
Testis weights relative to body weight were statistically significantly higher by ~11% only at 900 mg/
kg/day compared to the controls. However, males dosed at 900 mg/kg/day had a lower terminal b
ody weight and in view of the minor weight difference and no microscopic findings that could be corr
elated. This was therefore considered incidental.
Ovary weights were statistically significantly higher by ~19% (relative to body weight at 300 mg/kg/
day). However, no histologic findings could be correlated to this finding and no similar differences
were found in any other generation or cohort of animals, so this was considered incidental.
Thyroid/parathyroid gland weights were statistically significantly higher by ~21% (relative to body
weight) in female animals dosed at 900 mg/kg/day, compared to controls. However, no histologic fi
ndings could be correlated to this finding, thyroid hormone was not increased in a noteworthy fashio
n, and no similar differences were found in any other generation or cohort of animals, so this was c
onsidered incidental.
Seminal vesicle weights were statistically significantly higher by ~22% (relative to body weight) in
animals dosed at 900 mg/kg/day, compared to controls. However, no histologic findings could be co
rrelated to this finding, so this was considered not test item-related.
(Cohort 2A, 2B, F1 unselected pups, F2 unselected pups):
There were no test item-related organ weight differences. - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- Scheduled Euthanasia Animals - F1 Animals:
There were no test item-related gross findings.
Gross findings observed were of the nature commonly observed in this strain and age of rat, or occurred at a similar incidence in control and treated animals, and, therefore, were considered not to be test item-related.
Scheduled Euthanasia Animals (F1 Unselected pups):
There were no test item-related gross pathology findings.
The only abnormality noted was a dark red focus on the thymus in one male pup. Although the parents of these pups had been given Omnirad 184 at a dose of 900 mg/kg/day, this finding was considered not to be test item-related because it is a common agonal change. - Histopathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- (Cohort 1A)
In the liver, there was diffuse centrilobular hypertrophy of minimal to mild severity. For males only,
this was seen in all dosage groups, whereas for females this was only identified in animals dosed at
≥ 300 mg/kg/day. The incidence followed a dose-related distribution and was therefore considered
test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at ≥ 300 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184 were correlated to the histopat
hologic findings of centrilobular hypertrophy. This correlation was evident in males dosed at ≥ 100 mg/
kg/day and females at ≥ 300 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal seve
rity. This was characterised by an increase in the number of goblet cells. For both females and male
s, this was seen at ≥ 300 mg/kg/day. Although at a low incidence, this finding was only seen in anim
als treated with Omnirad 184 and in view of similar findings seen in the F0 generation, this finding was
considered test item-related in males and females dosed at ≥ 300 mg/kg/day.
Additional Microscopic Findings:
In the kidneys, there was a higher incidence of basophilic tubules with a minimal to mild severity, in
male animals given Omnirad 184 at a dose of 900 mg/kg/day. This was however a small difference
and although the males from F0 and F1 cohort 1B showed a similar difference, it was even less appa
rent at the highest dose level (900 mg/kg/day). Due to the fact that this lesion is a common backgro
und finding and occurred at a low severity, this was considered incidental.
In the liver of three animals given Omnirad 184 (two females dosed at 100 mg/kg/day and one female
dosed at 300 mg/kg/day), there was focal necrosis with subacute inflammation in the medial lobe
parenchyma, with a mild severity. Due to the low incidence, this finding was considered not test item related.
(Cohort 1B)
In the liver, there was diffuse centrilobular hypertrophy of minimal to mild severity. For males only, t
his was seen in all dosage groups, whereas for females this was only identified in animals dosed at
≥ 300 mg/kg/day. The overall incidence followed a dose related distribution and was therefore consid
ered test item-related in males dosed at ≥ 100 mg/kg/day and in females dosed at ≥ 300 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184, was correlated to the histop
athologic findings of centrilobular hypertrophy. This correlation was evident in males dosed at ≥ 100
mg/kg/day and females at ≥ 300 mg/kg/day. There were no test item-related microscopic findings in
the reproductive tissues.
(F1 Cohort 2A and 2B)
There were no test item-related microscopic findings. Microscopic findings occurred at similar incid
ences in control and treated animals, and, therefore, were considered not to be test item-related. - Other effects:
- no effects observed
- Description (incidence and severity):
- Acoustic startle:
There were no test item-related effects on maximum response amplitude, average response amplitude or time to maximum response in males or females at up to 900 mg/kg/day.
Estrous Cycles:
The number of days after vaginal opening before an estrus occurred was comparable with controls at all test item dose levels. There was no effect on the number or length of estrous cycles during the 2 weeks before necropsy at up to 900 mg/kg/day, compared with controls.
There was no effect on the number or length of estrous cycles during the 2 weeks before pairing or during the mating period at up to 900 mg/kg/day, compared with controls.
Mating Performance, Fertility and Duration of Gestation:
The following Cohort 1B females were euthanised on GD 24 as they had failed to produce litters and were found not to be pregnant; 1629F (control), 2624F, 2627F, 2628F and 2637F (100 mg/kg/day) and 3629F (300 mg/kg/day).
See table 22, table 24, and table 26
Pre- and Post-Implantation Loss:
There were no test item-related effects on number of corpora lutea, implant counts, pre- or post-implantation loss or total number of pups born at up to 900 mg/kg/day. - Behaviour (functional findings):
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test item-related effects on qualitative or quantitative functional observational para
meters at up to 900 mg/kg/day. - Developmental immunotoxicity:
- no effects observed
- Description (incidence and severity):
- Splenic Immunophenotyping Evaluation
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the immune cell populations in either the F0 adults rats analysed following administration of Omnirad 184 by oral gavage, or their resulting offspring. - Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- food consumption and compound intake
- clinical biochemistry
- organ weights and organ / body weight ratios
- histopathology: non-neoplastic
- Critical effects observed:
- no
- Clinical signs:
- not examined
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- There were no unscheduled deaths for F2 generation animals.
- Body weight and weight changes:
- not examined
- Food consumption and compound intake (if feeding study):
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- no effects observed
- Nipple retention in male pups:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- There were no test item-related organ weight differences.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no test item-related gross pathology findings.
Abnormalities noted at necropsy included a dark red focus on the thymus in one female pup. Although the parents of these pups had been given Omnirad 184 at a dose of 100 mg/kg/day, this finding was considered not to be test item-related because it is a common agonal change. In the liver of one male and three female pups whose parents had been treated with Omnirad 184 at a dose of 100 mg/kg/day there was mottled discoloration, abnormal appearance and a pale nodule on the medial lobe. Due to the low incidence and because no other animals were found with these gross lesions, they were considered not test item-related. - Histopathological findings:
- not examined
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- no effects observed
- Description (incidence and severity):
- Splenic Immunophenotyping Evaluation:
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the immune cell populations in either the F0 adults rats analysed following administration of Omnirad 184 by oral gavage, or their resulting offspring. - Key result
- Dose descriptor:
- NOAEL
- Generation:
- F2
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- viability
- sexual maturation
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Lowest effective dose / conc.:
- 900 mg/kg bw/day
- Treatment related:
- no
- Conclusions:
- The administration of Omnirad 184 once daily (or twice daily 3 hours apart on some occasions) via
oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no
test item-related deaths. There were occasional observations of decreased activity, abnormal, un
coordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose le
vel-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body we
ight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/
day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. Ther
e were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and
F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity
was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirme
d by overall ambulation values, or any associated clinical findings, detailed functional observations or
brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/
kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 male
s only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels
(up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/
kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values
or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target
organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertro
phy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney
weights were seen in both sexes. In neurological investigations, there were no test item-related brain
weight differences, gross or histological findings or effects on the brain dimensions. There were no
histological findings in the reproductive tissues of the females that had not been observed to litter tha
t would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1
adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the i
n-life and pathological findings were considered not to be adverse. For F1 and F2 developmental para
meters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on
the pup numbers, survival or development. - Executive summary:
The objective of this study was to determine the potential toxicity of the chemical Omnirad 184, when given by oral gavage to adult rats and their offspring. This study was designed to provide an evaluation of reproduction and development as well as a thorough evaluation of systemic toxicity in pregnant and lactating females and their offspring. Detailed examination of key developmental endpoints, such as offspring viability, neonatal health, developmental status at birth, physical and functional development until adulthood, nervous and immune system development was expected to identify any specific target organs in the offspring. In addition, the study provided information about the effects of Omnirad 184 on the integrity and performance of the adult male and female reproductive systems and a neurobehavioural assessment of the F1 generation.
The study design was as follows:
Text Table 1 Experimental Design – F0 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
25
25
2
Omnirad 184
100
10
10
10.03
100
25
25
3
Omnirad 184
300
10
30
30.09
301
25
25
4
Omnirad 184
900
10
90
90.27
903
25
25
a Dose volume was based on the most recent body weight measurement.
Control/vehicle = 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 2 Experimental Design – F1 Unselected PND (Post Natal Day) 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 3 Experimental Design – Cohort 1A (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Omnirad 184
100
10
10
10.03
100
20
20
3
Omnirad 184
300
10
30
30.09
301
20
21d
4
Omnirad 184
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and
450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
d One female given 300 mg/kg/day died within minutes of receiving its first dose on Day 1, and another female
was added to the group.Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 4 Experimental Design – Surplus F1 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Omnirad 184
100
10
10
10.03
100
10
10
3
Omnirad 184
300
10
30
30.09
301
10
10
4
Omnirad 184
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 5 Experimental Design – Cohort 1B (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Omnirad 184
100
10
10
10.03
100
20
20
3
Omnirad 184
300
10
30
30.09
301
20
20
4
Omnirad 184
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 6 Experimental Design – F2 Unselected PND 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 7 Experimental Design – Cohort 2A (Neurobehavioural)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Omnirad 184
100
10
10
10.03
100
10
10
3
Omnirad 184
300
10
30
30.09
301
10
10
4
Omnirad 184
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 8 Experimental Design – Cohort 2B (Neuropathology)a
Group No.
Number of Animals
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Cohort 2B were not dosed as they were terminated on PND 21/22.
In Week 17 of the study (F0 dosing and the start of F1 dosing), the 900 mg/kg/day dosing formulation could not be pulled up into the gavage tube being used for newly weaned pups, due to an unknown viscosity property, resulting in some subsets of F0 and/or F1 animals either not being dosed or being given the 300 mg/kg/day formulation either once or twice daily instead (refer to section 4.6.1 for details). From 19 Aug 2021, Group 4 F1 animals were dosed twice daily 3 hours apart at 450 mg/kg (equivalent to 900 mg/kg/day) until they were old enough for the next size gavage tube on 30 Aug 2021. Although there were occasions when animals were not dosed or received a dose level lower than that required by the study protocol, there was considered to be no overall impact on the results or study outcome due to the length of time that each generation was administered the test item dose levels correctly.
The following parameters and end points were evaluated in this study: clinical observations, body weights, food consumption, estrous cycles, mating performance, fertility indices, duration of gestation and overall litter performance, litter survival indices, litter and pup weights, pre-weaning physical development of F1 pups, assessments of sexual maturation of F1 animals, clinical pathology parameters (haematology, coagulation, clinical chemistry, and urinalysis), thyroid stimulating hormone (TSH) and thyroxine (T4) analysis, gross necropsy findings, splenic immunophenotyping analysis, organ weights, sperm evaluation, ovarian follicle counts and histopathological examinations.
None of the unscheduled deaths in any generation were considered to be test item-related.
For the F0 generation at 900 mg/kg/day, test item-related clinical observations of decreased activity, abnormal/uncoordinated gait and low carriage were occasionally seen for males and/or females before pairing, and decreased activity only during lactation. For F0 and F1 generations, salivation and/or ploughing were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, throughout the dosing periods. F0 and F1 males given 900 mg/kg/day had slightly lower body weight gain, and F0 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day.
There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development.
For males at 900 mg/kg/day, during the last 20 minutes of the 1 hour session, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this, the cause of this apparent reduction in motor activity could not be determined.
For F0 males given ≥300 mg/kg/day and F0 females given ≥100 mg/kg/day, triglycerides were lower by up to 0.75-fold, and for males only, cholesterol higher by up to 1.3-fold. For F1 animals at ≥100 mg/kg/day, triglycerides were lower than controls by up to 0.62-fold, and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold. There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day.
In the F0 animals dosed from 10 weeks prior to mating and continuing throughout mating and gestation until the females were at least LD 21, microscopically, centrilobular hypertrophy in the liver was considered test item-related in all dose groups in males but only at 900 mg/kg/day in females and mucosal hyperplasia of the glandular region, appearing as an increased number of goblet cells, was considered test item-related at ≥ 300 mg/kg/day in males and only at 900 mg/kg/day in females. Higher liver and kidney weights were seen in both sexes at all dose levels.
In the F1 cohort 1A animals, dosed from PND 21 until at least PND 90, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. In the stomach, there was mucosal hyperplasia of the glandular regions, appearing as an increased number of goblet cells. This finding was considered test item-related in males and females at ≥ 300 mg/kg/day. Liver weights were higher at ≥ 300 mg/kg/day in males and females. In addition, higher kidney weights were seen in females at all dose levels and at ≥ 300 mg/kg/day in males.
In the F1 cohort 1B animals, that were used to produce an F2 generation, and that were dosed from PND 21 until at least LD 21, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. Liver weights were higher at all dose levels in males and only at 900 mg/kg/day in females. In addition, higher kidney weights were seen in animals dosed at 900 mg/kg/day. Stomachs were not assessed microscopically in these animals. There were no test item-related organ weight differences in the female reproductive tissues.
There were no test item-related organ weight differences in a limited list of tissues in F1 and F2 pups that were euthanised at PND 21 without active dosing.
In the F1 cohort 2A and 2B animals used for neurological investigations (euthanised on PND 21 or dosed from PND 21 until at least PND 78-80), there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions.
There were no test item-related gross findings in the entire study, and there were only 6 premature deaths, all of which were considered not test item-related.
There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility.
In conclusion, administration of Omnirad 184 by once daily (or twice daily 3 hours apart on some occasions) oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no test item-related deaths. There were occasional observations of decreased activity, abnormal, uncoordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose level-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body weight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirmed by overall ambulation values, or any associated clinical findings, detailed functional observations or brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 males only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels (up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertrophy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney weights were seen in both sexes. In neurological investigations, there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions. There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1 adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the in-life and pathological findings were considered not to be adverse. For F1 and F2 developmental parameters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on the pup numbers, survival or development.
Referenceopen allclose all
Table 1 - Summary of Body Weights (g)
F0 Males
| Sex: Male | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 124.9 | 176.6 | 225.1 | 268.9 | 303.1 | 333 | 355.5 |
| Group 1 | SD | 16.6 | 18.5 | 21 | 23.8 | 24.7 | 26.3 | 27.3 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 123.8 | 174.3 | 225.7 | 270.7 | 305.6 | 333.2 | 355.7 |
| Group 2 | SD | 16.3 | 18.8 | 23.4 | 25.2 | 26.4 | 28.5 | 30.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.9 | -1.3 | 0.3 | 0.7 | 0.8 | 0.1 | 0.1 | |
| 300 mg/kg/day | Mean | 121 | 171.9 | 222.1 | 268.3 | 302.8 | 332.4 | 355.9 |
| Group 3 | SD | 15.1 | 18.9 | 21 | 23.1 | 22.1 | 24.6 | 26.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -3.1 | -2.7 | -1.3 | -0.2 | -0.1 | -0.2 | 0.1 | |
| 900 mg/kg/day | Mean | 124.9 | 178.2 | 230.5 | 277.9 | 310.1 | 334.8 | 352.2 |
| Group 4 | SD | 11.6 | 14.2 | 18.9 | 21.9 | 24.1 | 25.7 | 28.3 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0 | 0.9 | 2.4 | 3.4 | 2.3 | 0.5 | -0.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 43 | 50 | 57 | 64 | 71 | 78 | 85 | ||
| 0 mg/kg/day | Mean | 374.2 | 389 | 406.9 | 417.8 | 427.7 | 428 | 435.6 |
| Group 1 | SD | 30.2 | 31.6 | 33.7 | 35 | 34.9 | 33.2 | 34.4 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 374.4 | 392.8 | 408 | 417.3 | 423.9 | 427.8 | 437.4 |
| Group 2 | SD | 32.8 | 34.9 | 36.1 | 35.5 | 36.7 | 37.4 | 37.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0.1 | 1 | 0.3 | -0.1 | -0.9 | -0.1 | 0.4 | |
| 300 mg/kg/day | Mean | 373.7 | 390.5 | 404.8 | 415.6 | 424 | 426.6 | 436.5 |
| Group 3 | SD | 29.8 | 32.2 | 33.2 | 33.8 | 36.9 | 36.4 | 38.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | 0.4 | -0.5 | -0.5 | -0.9 | -0.3 | 0.2 | |
| 900 mg/kg/day | Mean | 365.9 | 378.8 | 393.4 | 403.8 | 411.4 | 415.7 | 423.1 |
| Group 4 | SD | 29.7 | 32 | 33.1 | 34.7 | 35.5 | 34.6 | 36.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -2.2 | -2.6 | -3.3 | -3.3 | -3.8 | -2.9 | -2.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 92 | 99 | 106 | 113 | 120 | 127 | 134 | ||
| 0 mg/kg/day | Mean | 441.8 | 453.6 | 466.3 | 472.8 | 483.5 | 489.2 | 496.7 |
| Group 1 | SD | 35.9 | 36.1 | 37.8 | 37.8 | 38.9 | 39 | 41.7 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 441.6 | 450.2 | 462.2 | 470.4 | 478 | 487.7 | 492.6 |
| Group 2 | SD | 37.1 | 37.6 | 39.7 | 40.7 | 41.1 | 43.8 | 45.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | -0.7 | -0.9 | -0.5 | -1.1 | -0.3 | -0.8 | |
| 300 mg/kg/day | Mean | 446.7 | 452.6 | 463.6 | 475.6 | 483.9 | 492.7 | 496.6 |
| Group 3 | SD | 40.7 | 42.3 | 44.7 | 47 | 49.7 | 50.9 | 53.1 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 1.1 | -0.2 | -0.6 | 0.6 | 0.1 | 0.7 | 0 | |
| 900 mg/kg/day | Mean | 434.1 | 442.2 | 449.5 | 458.4 | 464.4 | 475.5 | 478.6 |
| Group 4 | SD | 38 | 39.8 | 40.3 | 40.7 | 40.7 | 42.5 | 44.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -1.7 | -2.5 | -3.6 | -3 | -3.9 | -2.8 | -3.6 | |
| Sex: Male | Day(s) Relative to Start Date | ||||
| 137 | 138 | 141 | 142 | ||
| 0 mg/kg/day | Mean | 507.9 | 501.8 | 493.1 | 473.4 |
| Group 1 | SD | 58.2 | 33.5 | 36.1 | 43.3 |
| N | 8 - | 6 - | 11 - | 5 - | |
| 100 mg/kg/day | Mean | 488 | 512.6 | 490.8 | 477.7 |
| Group 2 | SD | 46.5 | 39.6 | 50.5 | 63.9 |
| N | 5 | 7 | 13 | 7 | |
| %Diff | -3.9 | 2.1 | -0.5 | 0.9 | |
| 300 mg/kg/day | Mean | 458 | 534.4 | 491 | 494.8 |
| Group 3 | SD | 26.4 | 57.8 | 46.4 | 52.9 |
| N | 4 | 8 | 13 | 8 | |
| %Diff | -9.8 | 6.5 | -0.4 | 4.5 | |
| 900 mg/kg/day | Mean | 473.8 | 494.5 | 480.2 | 471.2 |
| Group 4 | SD | 41.8 | 27.5 | 54.1 | 65.1 |
| N | 8 | 4 | 13 | 5 | |
| %Diff | -6.7 | -1.5 | -2.6 | -0.5 | |
Table 2 - Summary of Body Weights (g)
F0 Females - Prior to mating
| Sex: Female | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 72.9 | 110.5 | 136.4 | 155.4 | 173.6 | 188.9 | 197.4 |
| Group 1 | SD | 13.5 | 13.9 | 14 | 13.8 | 13.3 | 13.9 | 14.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 76 | 114.9 | 140.8 | 162 | 181.1 | 195.7 | 206.3 |
| Group 2 | SD | 10.2 | 11.1 | 11.6 | 13.5 | 14.4 | 14.5 | 16.5 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 3.9 | 3.3 | 4.3 | 4.3 | 3.6 | 4.5 | |
| 300 mg/kg/day | Mean | 74.9 | 114.2 | 142.2 | 162.9 | 183.5 | 198.8 | 209.1* |
| Group 3 | SD | 12.4 | 11.1 | 10.6 | 12.3 | 12.8 | 14.7 | 15.6 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 2.8 | 3.4 | 4.3 | 4.9 | 5.7 | 5.2 | 5.9 | |
| 900 mg/kg/day | Mean | 77.2 | 116.4 | 147.2* | 171.3** | 191.6** | 208.2** | 214.9** |
| Group 4 | SD | 14.1 | 14.3 | 15.3 | 17.2 | 17.4 | 17.9 | 18.2 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 5.9 | 5.3 | 7.9 | 10.3 | 10.3 | 10.2 | 8.9 |
| Sex: Female | Day(s) Relative to Start Date | |||||
| 43 | 50 | 57 | 64 | 71 | ||
| 0 mg/kg/day | Mean | 207.3 | 214.6 | 222.2 | 226.2 | 230.5 |
| Group 1 | SD | 14.9 | 13.1 | 14.8 | 14.4 | 15.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 216 | 225.6 | 229.6 | 232.8 | 240.8 |
| Group 2 | SD | 17.1 | 18.4 | 19 | 20.5 | 20.5 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 5.2 | 3.3 | 2.9 | 4.5 | |
| 300 mg/kg/day | Mean | 217.2 | 227.9* | 234.7* | 238.4 | 245.9* |
| Group 3 | SD | 16.5 | 18.6 | 19.4 | 18.9 | 19 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.8 | 6.2 | 5.6 | 5.4 | 6.7 | |
| 900 mg/kg/day | Mean | 227.0** | 233.0** | 242.0** | 244.5** | 250.5** |
| Group 4 | SD | 17.2 | 17.5 | 18.6 | 19 | 19.7 |
| N | 24 | 24 | 24 | 24 | 24 | |
| %Diff | 9.5 | 8.6 | 8.9 | 8.1 | 8.7 |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 3 - Summary of Body Weights (g)
F0 Females - Gestation
| Sex: Female | Day(s) Relative to Mating (Litter: A) | ||||
| 0 | 7 | 14 | 20 | ||
| 0 mg/kg/day | Mean | 222.8 | 246.1 | 269.9 | 325.9 |
| Group 1 | SD | 14.5 | 13.5 | 16.3 | 24 |
| N | 18 - | 18 - | 18 - | 18 - | |
| 100 mg/kg/day | Mean | 237.2* | 259.5 | 282.3 | 338.5 |
| Group 2 | SD | 20.1 | 20.5 | 22.9 | 31.2 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 6.5 | 5.5 | 4.6 | 3.9 | |
| 300 mg/kg/day | Mean | 242.1** | 262.4* | 284.2 | 343.6 |
| Group 3 | SD | 20.5 | 22.4 | 24.6 | 28.1 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 8.6 | 6.6 | 5.3 | 5.4 | |
| 900 mg/kg/day | Mean | 243.7** | 264.5* | 291.1* | 352.5* |
| Group 4 | SD | 17.2 | 21.1 | 23.4 | 34.4 |
| N | 22 | 22 | 22 | 22 | |
| %Diff | 9.4 | 7.5 | 7.9 | 8.2 | |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 4 - Summary of Body Weights (g)
F0 Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | ||
| Sex | 7 | ||||||
| 1F | Mean | 261.7 | 274 | 275.1 | 282.8 | 280.1 | |
| SD | 19.1 | 20.8 | 19.3 | 21.2 | 16.6 | ||
| N | 23 | 22 | 23 | 23 | 23 | ||
| Mean | 268.2 | 280.9 | 283.3 | 290.9 | 288 | ||
| 2F | SD | 22.1 | 22.6 | 23 | 23.6 | 21.7 | |
| N | 23 | 24 | 24 | 24 | 24 | ||
| %Diff G1 | 2.5 | 2.5 | 3 | 2.9 | 2.8 | ||
| Mean | 270.8 | 282.7 | 285.6 | 294.8 | 291 | ||
| SD | 25.7 | 25 | 24.4 | 23.4 | 19.9 | ||
| 3F | N | 25 | 25 | 25 | 25 | 25 | |
| %Diff G1 | 3.5 | 3.2 | 3.8 | 4.2 | 3.9 | ||
| Mean | 271.6 | 286.4 | 292.2 | 302.6a | 300.1b | ||
| 4F | SD | 23.4 | 24 | 24.8 | 25.7 | 25.2 | |
| N | 22 | 22 | 22 | 22 | 22 | ||
| %Diff G1 | 3.8 | 4.5 | 6.2 | 7 | 7.1 | ||
Significantly different from control group 1 value :a=p≤0.05,b=p≤0.01 (Dunnett)
Table 5 - Summary of Body Weights (g)
F1 Animals - Cohort 1A
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
| 0 mg/kg/day | Mean | 56 | - | 71.8 | 119.9 | 172.1 | 219.5 | 269.6 |
| Group 1 | SD | 5.1 | - | 8.8 | 12.2 | 15.8 | 19.7 | 23.4 |
| N | 20 - | - | 20 - | 20 - | 20 - | 20 - | 20 - | |
| - | ||||||||
| 100 mg/kg/day | Mean | 56.3 | 61.0n | 70.8 | 116.4 | 166.9 | 213.7 | 261.2 |
| Group 2 | SD | 6.6 | 6.6 | 9 | 13.6 | 16.5 | 20.5 | 26.4 |
| N | 19 | 3 - | 18 | 20 | 20 | 20 | 20 | |
| %Diff | 0.7 | -1.4 | -2.9 | -3 | -2.6 | -3.1 | ||
| 300 mg/kg/day | Mean | 57.9 | 76.0n | 76.8 | 126.3 | 179.8 | 227 | 276.8 |
| Group 3 | SD | 4.9 | - | 9.3 | 14 | 16.9 | 19.6 | 24.9 |
| N | 20 | 1 - | 19 | 20 | 20 | 20 | 20 | |
| %Diff | 3.5 | 6.9 | 5.4 | 4.5 | 3.4 | 2.7 | ||
| 900 mg/kg/day | Mean | 54.7 | 59.0n 4.3 | 70.6 | 115.8 | 167.5 | 212.9 | 258.5 |
| Group 4 | SD | 5.5 | 6 - | 8.2 | 12.5 | 16.5 | 21.7 | 25.5 |
| N | 15 | 19 | 20 | 20 | 20 | 20 | ||
| %Diff | -2.2 | -1.6 | -3.4 | -2.7 | -3 | -4.1 |
[G] - Anova & Dunnett [I] - n - Inappropriate for statistics
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 41 | 48 | 55 | 62 | 68 | 69 | 70 | ||
| 0 mg/kg/day | Mean | 309.2 | 343.6 | 368.2 | 390.7 | 408.1 | 394 | 405.5 |
| Group 1 | SD | 26.7 | 31.5 | 33 | 37 | 40.1 | 42.2 | 40.7 |
| N | 20 - | 20 - | 20 - | 20 - | 14 - | 6 - | 20 - | |
| 100 mg/kg/day | Mean | 300.1 | 335.3 | 360.5 | 382.8 | 393 | 408.2 | 401.6 |
| Group 2 | SD | 29.4 | 32.6 | 35.7 | 36.5 | 35.3 | 42.7 | 34.3 |
| N | 20 | 20 | 20 | 20 | 16 | 6 | 16 | |
| %Diff | -2.9 | -2.4 | -2.1 | -2 | -3.7 | 3.6 | -1 | |
| 300 mg/kg/day | Mean | 317.3 | 349.4 | 376.1 | 396.8 | 411.1 | 445.9 | 411.1 |
| Group 3 | SD | 28 | 33 | 37.6 | 38.7 | 46.2 | 36.6 | 36.1 |
| N | 20 | 20 | 20 | 20 | 14 | 7 | 19 | |
| %Diff | 2.6 | 1.7 | 2.1 | 1.6 | 0.8 | 13.2 | 1.4 | |
| 900 mg/kg/day | Mean | 299.2 | 325.5 | 346.4 | 363.5 | 386.8 | 381.7 | 382.7 |
| Group 4 | SD | 29 | 32.1 | 37.5 | 40 | 40.6 | 48.2 | 38.9 |
| N | 20 | 20 | 20 | 20 | 14 | 11 | 15 | |
| %Diff | -3.2 | -5.3 | -5.9 | -7 | -5.2 | -3.1 | -5.6 |
Sex: Male |
| Day(s) Relative to Start Date |
71 | ||
0 mg/kg/day
Group 1 | Mean SD N
| - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 447.0n 69.3 2 - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | - - - - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | - - - - |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 4.9 20 - | - - - - | 66.0 7.8 20 - | 103.8 9.7 20 - | 136.5 10.7 20 - | 157.2 10.2 20 - | 176.5 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 55.3 4.9 19 5.3 | 58.0n 3.0 3 - | 68.9 5.7 18 4.5 | 106.1 9.4 20 2.2 | 138.3 11.9 20 1.3 | 159.7 14.1 20 1.6 | 178.8 16.8 20 1.3 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.0 5.0 21 4.7 | 71.0n - 1 - | 69.4 8.9 19 5.2 | 105.5 11.3 20 1.7 | 138.6 11.6 20 1.5 | 160.2 12.3 20 1.9 | 179.9 13.8 20 1.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 52.1 4.8 15 -0.8 | 56.3n 6.0 7 - | 66.1 8.2 18 0.2 | 103.2 10.6 20 -0.6 | 138.6 12.8 20 1.5 | 159.9 15.9 20 1.7 | 182.0 15.5 20 3.1 |
[G] - Anova & Dunnett
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 [G] | 48 [G] | 55 [G] | 62 [G] | 68 [G] | 69 [G] | 70 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.0 12.1 20 - | 203.2 14.4 20 - | 212.2 15.5 20 - | 221.0 18.8 20 - | 231.6 18.8 13 - | 228.1 18.9 7 - | 245.3n 11.0 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 192.4 18.5 20 1.8 | 205.7 17.3 20 1.2 | 214.1 18.0 20 0.9 | 224.2 20.5 20 1.4 | 235.2 22.6 17 1.6 | 216.7 6.8 3 -5.0 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 194.4 13.6 20 2.8 | 206.5 14.6 20 1.6 | 216.4 16.9 20 2.0 | 226.1 17.7 20 2.3 | 231.5 16.3 14 0.0 | 242.3 12.6 6 6.2 | 220.0n - 1 -10.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 201.6 17.1 20 6.7 | 209.2 17.6 20 3.0 | 218.0 17.7 20 2.7 | 226.5 18.3 20 2.5 | 237.5 19.1 15 2.6 | 235.6 26.0 5 3.3 | - - - - |
[G] - Anova & Dunnett
Sex: Female |
| Day(s) Relative to Start Date |
| ||
71 [G] | 72 [I] | 73 [G] | 74 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 226.9 16.6 8 - | - - - - | 228.6 21.1 5 - | 228.8 17.9 4 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 228.3 29.4 8 0.6 | 223.7n 12.2 3 - | 233.8 11.4 5 2.3 | 233.8 25.4 4 2.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 233.1 15.5 9 2.7 | 268.0n - 1 - | 225.0n 11.3 2 -1.6 | 237.7 20.5 7 3.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 235.0 23.9 10 3.6 | 247.0n 11.3 2 - | 232.8 19.2 6 1.9 | 235.0n 25.5 2 2.7 |
[G] - Anova & Dunnett: n - Inappropriate for statistics
Table 7 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Males
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 5.4 20 - | - - - - | 70.5 8.5 20 - | 116.5 12.0 20 - | 167.7 13.3 20 - | 212.9 17.3 20 - | 261.7 20.9 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 57.4 5.2 19 5.1 | 60.0n 8.5 3 - | 72.9 8.7 18 3.5 | 117.5 14.9 20 0.8 | 166.8 20.5 20 -0.6 | 211.2 25.9 20 -0.8 | 258.5 29.7 20 -1.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 56.8 5.7 20 4.0 | 70.0n - 1 - | 74.9 10.8 19 6.2 | 124.1 15.9 20 6.5 | 175.7 18.3 20 4.7 | 222.6 21.4 20 4.6 | 271.2 25.8 20 3.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 54.5 4.5 15 -0.1 | 59.3n 5.9 7 - | 69.9 8.7 18 -0.8 | 115.4 10.8 20 -0.9 | 166.8 12.3 20 -0.5 | 212.2 15.2 20 -0.3 | 258.6 18.7 20 -1.2 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 300.5 25.6 20 - | 334.6 30.5 20 - | 358.4 34.8 20 - | 381.5 38.4 20 - | 395.5 41.5 20 - | 410.3 44.0 20 - | 421.2 46.7 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 297.5 32.5 20 -1.0 | 330.9 34.9 20 -1.1 | 357.3 37.9 20 -0.3 | 379.6 39.8 20 -0.5 | 399.5 39.4 20 1.0 | 414.5 41.7 20 1.0 | 429.9 42.0 20 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 310.2 26.4 20 3.2 | 342.6 29.7 20 2.4 | 370.5 31.5 20 3.4 | 393.0 33.5 20 3.0 | 413.3 33.9 20 4.5 | 427.1 37.4 20 4.1 | 438.9 39.0 20 4.2 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 297.9 23.2 20 -0.8 | 324.7 26.1 20 -2.9 | 348.3 30.2 20 -2.8 | 365.3 33.5 20 -4.2 | 383.9 36.0 20 -2.9 | 399.6 35.5 20 -2.6 | 410.4 40.6 20 -2.6 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
90 | 97 | 104 | 111 | 118 | 125 | 128 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 431.7 46.1 20 - | 434.7 44.9 20 - | 441.9 43.5 20 - | 453.5 48.3 20 - | 464.3 49.9 20 - | 473.8 52.6 20 - | 484.1 58.9 8 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 440.9 44.6 20 2.1 | 444.3 45.0 20 2.2 | 452.5 46.6 20 2.4 | 463.5 46.1 20 2.2 | 477.7 47.1 20 2.9 | 492.3 48.2 20 3.9 | 494.2 58.5 11 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 448.1 41.2 20 3.8 | 452.0 38.8 20 4.0 | 456.2 39.7 20 3.2 | 471.1 40.7 20 3.9 | 483.8 43.3 20 4.2 | 494.5 45.3 20 4.4 | 515.1 42.9 9 6.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 415.5 40.8 20 -3.8 | 424.9 41.7 20 -2.3 | 428.4 41.3 20 -3.1 | 442.0 42.1 20 -2.5 | 451.7 42.8 20 -2.7 | 460.8 41.2 20 -2.8 | 463.5 45.3 12 -4.3 |
Sex: Male |
|
| Day(s) Relative to Start Date |
|
129 [G] | 130 [G] | 131 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 465.0 62.2 6 - | 475.3 45.0 9 - | - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 504.9 36.5 7 8.6 | 502.0 45.0 7 5.6 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 514.8 41.9 5 10.7 | 471.0 41.8 8 -0.9 | 485.5n 36.8 4 - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 476.3 43.3 8 2.4 | 436.3 10.1 3 -8.2 | 529.0n - 1 - |
Table 8 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Prior to Mating
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 5.1 20 - | - - - - | 66.2 7.1 20 - | 103.6 9.0 20 - | 136.1 9.5 20 - | 156.9 10.6 20 - | 174.8 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 53.8 7.9 19 2.4 | 57.3n 4.0 3 - | 66.6 10.4 18 0.7 | 103.4 13.9 20 -0.1 | 133.7 13.8 20 -1.7 | 155.3 14.0 20 -1.0 | 173.5 14.1 20 -0.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 4.4 20 6.0 | 68.0n - 1 - | 71.6 7.5 19 8.3 | 109.3 8.1 20 5.5 | 143.2 9.6 20 5.3 | 164.1 10.0 20 4.6 | 180.9 11.8 20 3.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 51.3 4.3 15 -2.3 | 56.3n 6.6 7 - | 64.6 7.4 18 -2.4 | 102.8 10.3 20 -0.7 | 136.6 9.8 20 0.4 | 157.6 13.1 20 0.4 | 176.8 13.7 20 1.1 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.6 11.3 20 - | 204.2 11.0 20 - | 210.9 11.8 20 - | 218.6 12.6 20 - | 227.6 11.8 20 - | 232.5 11.5 20 - | 236.2 12.5 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 188.1 15.8 20 -0.8 | 200.3 15.1 20 -1.9 | 209.8 15.4 20 -0.5 | 218.7 14.7 20 0.1 | 226.3 16.5 20 -0.6 | 230.7 18.2 20 -0.8 | 232.7 17.3 20 -1.5 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 199.2 12.4 20 5.1 | 211.3 13.2 20 3.5 | 220.1 14.6 20 4.4 | 227.0 15.2 20 3.9 | 236.9 15.5 20 4.1 | 243.1 16.4 20 4.5 | 246.7 17.5 20 4.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 195.6 14.1 20 3.2 | 203.7 14.7 20 -0.2 | 212.2 15.4 20 0.6 | 219.5 16.9 20 0.4 | 231.0 16.5 20 1.5 | 236.4 18.1 20 1.7 | 237.4 17.1 20 0.5 |
Sex: Female |
| Day(s) Relative to Start Date |
90 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 240.0 13.4 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 239.6 16.7 20 -0.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 251.2 17.1 20 4.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 242.7 18.8 20 1.1 |
Table 9 - Summary of Body Weights (g)
F1 Animals - Cohort 1B - Gestation
Sex: Female |
|
| Day(s) Relative to Mating (Litter: A) |
| |
0 | 7 | 14 | 20 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 241.9 12.7 19 - | 261.8 15.3 19 - | 284.1 15.0 19 - | 337.5 18.9 19 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 238.8 14.3 16 -1.3 | 258.2 14.3 16 -1.4 | 277.9 14.9 16 -2.2 | 334.6 19.9 16 -0.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 253.5 16.6 19 4.8 | 271.6 16.9 19 3.8 | 291.6 16.6 19 2.6 | 348.9 19.3 19 3.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 241.1 16.7 20 -0.3 | 260.8 20.7 20 -0.4 | 278.0 23.3 20 -2.1 | 336.5 32.1 20 -0.3 |
Table 10 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | |
| Sex | 7 | |||||
| 1F | Mean | 264.2 | 268.9 | 276.9 | 290.6 | 284.2 |
| SD | 14.4 | 12.9 | 14.9 | 16.8 | 17.9 | |
| N | 18 | 17 | 18 | 18 | 18 | |
| Mean | 262.3 | 271.3 | 275.8 | 288.6 | 286.3 | |
| 2F | SD | 14.5 | 15 | 14.2 | 16.6 | 15.4 |
| N | 16 | 15 | 16 | 16 | 16 | |
| %Diff G1 | -0.7 | 0.9 | -0.4 | -0.7 | 0.8 | |
| Mean | 278.4a | 288.5b | 292.6a | 306.2a | 302.1a | |
| SD | 18.9 | 16.6 | 15.7 | 15.6 | 16.2 | |
| 3F | N | 19 | 19 | 19 | 19 | 19 |
| %Diff G1 | 5.4 | 7.3 | 5.6 | 5.4 | 6.3 | |
| Mean | 265.9 | 276.3 | 283.6 | 292.4 | 291.5 | |
| 4F | SD | 21.6 | 24.5 | 20.3 | 21.7 | 22.9 |
| N | 20 | 19 | 20 | 20 | 19 | |
| %Diff G1 | 0.6 | 2.7 | 2.4 | 0.6 | 2.6 |
Table 11 - Summary of Body Weights (g)
F1 Animals - Cohort 2A
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 4.7 10 - | - - - - | 68.4 8.0 10 - | 115.1 11.9 10 - | 169.5 12.1 10 - | 219.4 13.6 10 - | 269.5 18.2 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.6 5.5 9 -0.1 | 59.0n 7.1 2 - | 67.0 8.9 9 -2.0 | 111.0 12.0 10 -3.6 | 160.7 12.0 10 -5.2 | 205.3 14.3 10 -6.4 | 250.8 18.2 10 -6.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 58.5 5.9 10 7.1 | 76.0n - 1 - | 74.7 10.9 9 9.2 | 122.9 14.3 10 6.8 | 177.1 16.3 10 4.5 | 224.4 19.6 10 2.3 | 273.9 23.0 10 1.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 55.4 3.6 9 1.5 | 59.0n 7.1 2 - | 67.3 9.2 9 -1.6 | 112.4 11.1 10 -2.3 | 164.3 12.4 10 -3.1 | 211.4 15.1 10 -3.6 | 259.5 21.4 10 -3.7 |
Sex: Male |
|
|
| Day(s) Relative to Start Date |
|
| |
41 | 48 | 55 | 58 | 59 | 60 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 308.0 21.9 10 - | 342.0 24.0 10 - | 368.4 25.7 10 - | 369.8 24.4 4 - | 396.8 32.0 5 - | 405.0 37.8 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 287.9 19.6 10 -6.5 | 320.4 24.2 10 -6.3 | 347.9 26.8 10 -5.6 | 376.8 42.4 4 1.9 | 369.8 20.2 6 -6.8 | 360.5 16.2 4 -11.0 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 313.5 26.9 10 1.8 | 348.1 27.9 10 1.8 | 372.8 25.9 10 1.2 | 403.3 32.3 6 9.1 | 404.0 54.1 3 1.8 | 369.5 14.5 4 -8.8 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 298.6 22.8 10 -3.1 | 326.5 27.2 10 -4.5 | 347.3 31.0 10 -5.7 | 362.0 46.0 4 -2.1 | 376.9 25.2 7 -5.0 | 341.5n 0.7 2 -15.7 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 51.5 4.2 10 - | - - - - | 62.9 6.5 10 - | 98.9 9.8 10 - | 128.6 9.3 10 - | 149.3 8.6 10 - | 166.7 9.1 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.2 2.9 9 5.3 | 54.0n 2.8 2 - | 65.4 5.5 9 4.0 | 100.5 8.6 10 1.6 | 131.3 8.2 10 2.1 | 155.1 8.6 10 3.9 | 172.1 10.8 10 3.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 7.4 10 8.2 | 76.0n - 1 - | 68.8 10.0 9 9.3 | 108.9 13.2 10 10.1 | 145.1** 12.9 10 12.8 | 167.6** 15.5 10 12.3 | 186.0** 18.3 10 11.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 53.6 3.7 9 4.0 | 57.0n 5.7 2 - | 63.2 5.3 9 0.5 | 100.5 6.2 10 1.6 | 135.7 10.3 10 5.5 | 159.2 10.4 10 6.6 | 179.3 12.7 10 7.6 |
Sex: Female |
|
| Day(s) Relative to Start Date |
|
| ||
41 [G] | 48 [G] | 55 [G] | 58 [G] | 59 [G1] | 60 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 179.8 10.1 10 - | 193.9 9.9 10 - | 203.7 10.7 10 - | 204.8 13.1 4 - | 214.0 12.7 5 - | 214.3 12.2 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 185.7 12.3 10 3.3 | 199.7 13.1 10 3.0 | 210.3 14.1 10 3.2 | 206.8 12.0 4 1.0 | 218.0 20.1 6 1.9 | 210.8 12.0 4 -1.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 202.3** 20.4 10 12.5 | 215.0** 20.3 10 10.9 | 223.8* 23.4 10 9.9 | 237.2 33.5 6 15.8 | 247.0 47.1 3 15.4 | 223.5 14.2 4 4.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 198.6* 14.7 10 10.5 | 210.7* 14.0 10 8.7 | 218.6 13.4 10 7.3 | 223.0 22.0 4 8.9 | 229.9 10.5 7 7.4 | 220.7 6.0 3 3.0 |
Tables 12. Summary of Clinical Chemistry Values
F0 Animals - Males. Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G1] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 71.4 12.3 10 - | 51.0 11.7 10 - | 128.5 12.4 10 - | 1.5 0.0 10 - | 217.1 68.4 10 - | 1.25 0.00 10 - | 7.35 0.66 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 74.6 10.3 9 1.04 | 49.1 12.0 9 0.96 | 126.0 25.8 9 0.98 | 1.5 0.0 9 1.00 | 292.7 163.0 9 1.35 | 1.25 0.00 9 1.00 | 6.90 0.68 9 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 60.5 * 4.9 10 0.85 | 44.5 7.8 10 0.87 | 125.1 32.5 10 0.97 | 1.5 0.0 10 1.00 | 213.2 70.8 10 0.98 | 1.25 0.00 10 1.00 | 7.16 0.35 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.6 * 5.4 10 0.86 | 36.8 14.1 9 0.72 | 115.4 28.5 10 0.90 | 1.5 0.0 10 1.00 | 196.8 60.0 10 0.91 | 1.25 0.00 10 1.00 | 6.94 0.68 10 0.94 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.2 3.2 10 - | 8.251 0.699 10 - | 1.789 0.249 10 - | 1.949 0.657 10 - | 66.29 1.89 10 - | 40.36 1.53 10 - | 25.93 2.18 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 32.1 3.5 9 0.94 | 8.778 1.195 9 1.06 | 1.853 0.393 9 1.04 | 1.933 0.681 9 0.99 | 66.09 3.31 9 1.00 | 41.22 2.83 9 1.02 | 24.87 2.61 9 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 * 2.9 10 0.87 | 8.113 0.521 10 0.98 | 2.170 * 0.404 10 1.21 | 1.594 0.199 10 0.82 | 66.67 2.46 10 1.01 | 41.93 1.91 10 1.04 | 24.74 2.11 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 31.5 3.6 10 0.92 | 8.208 0.634 10 0.99 | 2.327 ** 0.270 10 1.30 | 1.457 0.567 10 0.75 | 68.21 2.50 10 1.03 | 43.17 * 2.11 10 1.07 | 25.04 2.86 10 0.97 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.58 0.18 10 - | 2.618 0.059 10 - | 141.1 1.3 10 - | 4.98 0.29 10 - | 103.7 1.2 10 - | 1.404 0.193 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.69 0.22 9 1.07 | 2.637 0.076 9 1.01 | 141.4 1.9 9 1.00 | 4.90 0.34 9 0.98 | 103.7 1.4 9 1.00 | 1.531 0.178 9 1.09 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.71 0.17 10 1.08 | 2.654 0.055 10 1.01 | 140.3 2.2 10 0.99 | 4.97 0.15 10 1.00 | 102.3 1.8 10 0.99 | 1.545 0.152 10 1.10 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.75 0.28 10 1.11 | 2.716 ** 0.069 10 1.04 | 141.3 1.7 10 1.00 | 5.02 0.29 10 1.01 | 101.4 ** 2.0 10 0.98 | 1.591 0.187 10 1.13 |
Tables 13. Summary of Clinical Chemistry Values
F0 Animals - Females. Day: 115 Relative to Start Date
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 131.2 28.7 9 - | 117.1 17.5 9 - | 223.4 49.2 9 - | 1.5 0.0 9 - | 630.0 384.5 9 - | 1.25 0.00 9 - | 10.50 1.01 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 105.3 10.9 8 0.80 | 110.0 18.4 8 0.94 | 200.3 73.8 7 0.90 | 1.5 0.0 8 1.00 | 356.8 132.6 8 0.57 | 1.25 0.00 8 1.00 | 10.11 0.94 8 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 116.1 21.8 9 0.88 | 105.7 14.1 9 0.90 | 157.1 38.5 9 0.70 | 1.5 0.0 9 1.00 | 614.7 356.8 9 0.98 | 1.25 0.00 9 1.00 | 9.38 0.84 9 0.89 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 106.4 25.3 10 0.81 | 100.4 21.0 10 0.86 | 173.2 49.5 10 0.78 | 1.5 0.0 9 1.00 | 428.3 244.8 9 0.68 | 1.25 0.00 9 1.00 | 10.57 1.26 9 1.01 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G1] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 29.4 3.2 9 - | 4.740 1.150 9 - | 2.828 0.449 9 - | 0.589 0.061 9 - | 56.30 2.58 9 - | 40.21 1.79 9 - | 16.09 2.22 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 24.6 * 4.2 8 0.84 | 5.008 0.434 8 1.06 | 2.641 0.493 8 0.93 | 0.461 0.140 8 0.78 | 55.21 2.41 8 0.98 | 39.51 2.62 8 0.98 | 15.70 0.72 8 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 24.0 * 2.5 8 0.82 | 4.863 0.808 8 1.03 | 2.889 0.655 9 1.02 | 0.535 0.177 8 0.91 | 56.38 2.85 8 1.00 | 40.59 2.45 9 1.01 | 16.19 1.63 8 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 25.6 4.6 9 0.87 | 5.003 0.815 9 1.06 | 2.827 0.925 9 1.00 | 0.479 0.115 9 0.81 | 58.48 3.36 9 1.04 | 42.81 2.93 10 1.06 | 15.68 1.98 9 0.97 |
Sex: Female |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.53 0.34 9 - | 2.680 0.100 9 - | 140.7 2.1 9 - | 5.12 0.42 9 - | 99.8 2.2 9 - | 1.708 0.538 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.54 0.23 8 1.00 | 2.621 0.139 8 0.98 | 140.0 3.5 8 1.00 | 5.05 0.57 8 0.99 | 100.1 1.9 8 1.00 | 1.649 0.518 8 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.50 0.29 8 0.99 | 2.694 0.096 9 1.01 | 140.1 1.3 9 1.00 | 5.24 0.62 9 1.02 | 99.3 1.7 9 1.00 | 1.538 0.510 8 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.78 0.47 9 1.10 | 2.781 0.183 9 1.04 | 140.1 2.3 10 1.00 | 5.19 0.42 10 1.01 | 97.8 2.9 10 0.98 | 1.949 0.766 9 1.14 |
Tables 14. Summary of Clinical Chemistry Values
F1 Animals - Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 68.5 6.0 10 - | 57.6 11.4 10 - | 153.4 23.5 10 - | 1.5 0.0 9 - | 239.9 42.0 9 - | 1.25 0.00 9 - | 7.02 0.68 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 63.0 7.2 10 0.92 | 52.0 7.9 10 0.90 | 153.5 23.8 10 1.00 | 1.5 0.0 10 1.00 | 239.2 101.7 10 1.00 | 1.25 0.00 10 1.00 | 6.32 0.62 10 0.90 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 61.3 * 5.3 10 0.89 | 49.9 8.6 10 0.87 | 143.2 27.9 10 0.93 | 1.5 0.0 10 1.00 | 210.5 55.4 10 0.88 | 1.25 0.00 10 1.00 | 6.84 0.59 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.2 * 4.0 10 0.89 | 42.5 ** 13.0 10 0.74 | 141.5 23.9 10 0.92 | 1.5 0.0 10 1.00 | 174.9 37.4 10 0.73 | 1.25 0.00 10 1.00 | 7.24 0.85 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 30.8 4.0 9 - | 9.268 0.551 9 - | 1.723 0.237 9 - | 2.458 1.001 9 - | 66.48 1.47 9 - | 41.30 1.39 10 - | 25.16 1.24 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 28.1 3.8 10 0.91 | 8.529 ** 0.254 10 0.92 | 1.894 0.316 10 1.10 | 2.030 0.674 10 0.83 | 66.44 1.28 10 1.00 | 41.71 2.18 10 1.01 | 24.73 2.00 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 2.6 10 0.97 | 8.735 0.590 10 0.94 | 2.007 0.325 10 1.16 | 1.784 0.635 10 0.73 | 65.87 2.39 10 0.99 | 41.11 2.06 10 1.00 | 24.76 2.03 10 0.98 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.6 3.7 10 1.06 | 8.617 * 0.503 10 0.93 | 2.147 * 0.325 10 1.25 | 1.522 0.615 10 0.62 | 65.97 1.76 10 0.99 | 42.49 1.31 10 1.03 | 23.48 1.78 10 0.93 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G1] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.64 0.10 9 - | 2.592 0.033 9 - | 141.0 0.8 10 - | 5.09 0.30 10 - | 100.5 0.5 10 - | 1.747 0.191 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.70 0.19 10 1.03 | 2.620 0.064 10 1.01 | 142.0 1.2 10 1.01 | 5.12 0.30 10 1.01 | 101.7 * 0.8 10 1.01 | 2.034 ** 0.122 10 1.16 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.67 0.19 10 1.02 | 2.626 0.074 10 1.01 | 141.1 1.5 10 1.00 | 4.99 0.20 10 0.98 | 101.3 1.2 10 1.01 | 1.968 ** 0.179 10 1.13 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.83 0.17 10 1.11 | 2.657 0.064 10 1.02 | 141.6 2.0 10 1.00 | 4.95 0.18 10 0.97 | 99.9 1.6 10 0.99 | 2.065 ** 0.116 10 1.18 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 78.0 10.7 7 - | 55.9 14.4 7 - | 100.6 26.1 7 - | 1.5 0.0 7 - | 295.7 114.0 7 - | 1.25 0.00 7 - | 8.09 1.15 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 71.9 7.5 9 0.92 | 54.6 17.9 9 0.98 | 93.2 23.3 9 0.93 | 1.5 0.0 9 1.00 | 242.9 83.5 9 0.82 | 1.25 0.00 9 1.00 | 7.43 0.78 9 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 68.3 6.1 10 0.88 | 49.9 13.2 10 0.89 | 94.3 24.1 10 0.94 | 1.5 0.0 10 1.00 | 259.2 98.3 10 0.88 | 1.25 0.00 10 1.00 | 7.70 0.89 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 73.7 9.9 10 0.94 | 59.8 26.8 10 1.07 | 135.4 38.1 10 1.35 | 2.2 1.5 10 1.47 | 296.8 105.5 10 1.00 | 1.25 0.00 10 1.00 | 7.66 1.24 10 0.95 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.0 3.5 7 - | 8.577 0.866 7 - | 1.589 0.267 7 - | 1.421 0.304 7 - | 65.96 2.49 7 - | 45.56 2.60 7 - | 20.40 2.41 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 34.8 4.5 9 1.02 | 7.801 0.566 9 0.91 | 1.512 0.297 9 0.95 | 1.021 0.261 9 0.72 | 67.76 2.46 9 1.03 | 46.70 1.56 9 1.03 | 21.06 1.80 9 1.03 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 32.7 4.0 10 0.96 | 8.161 0.464 10 0.95 | 1.647 0.235 10 1.04 | 1.268 0.529 10 0.89 | 68.21 2.48 10 1.03 | 46.77 2.62 10 1.03 | 21.44 2.18 10 1.05 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.4 4.5 10 0.95 | 8.082 0.391 10 0.94 | 1.913 * 0.234 10 1.20 | 0.978 0.268 10 0.69 | 67.82 2.61 10 1.03 | 46.97 2.19 10 1.03 | 20.85 1.87 10 1.02 |
Sex: Female |
|
| Reporting Biochemistry |
|
| ||
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.27 0.35 7 - | 2.574 0.035 7 - | 140.3 1.1 7 - | 4.43 0.42 7 - | 103.6 1.4 7 - | 1.227 0.104 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.24 0.21 9 0.99 | 2.581 0.060 9 1.00 | 140.2 1.2 9 1.00 | 4.78 0.29 9 1.08 | 102.9 1.2 9 0.99 | 1.436 0.206 9 1.17 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.18 0.30 10 0.96 | 2.591 0.053 10 1.01 | 140.6 0.8 10 1.00 | 4.45 0.20 10 1.00 | 104.1 1.3 10 1.01 | 1.379 0.205 10 1.12 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.30 0.28 10 1.01 | 2.664 ** 0.039 10 1.03 | 140.4 1.1 10 1.00 | 4.86 * 0.27 10 1.10 | 102.2 0.9 10 0.99 | 1.312 0.263 10 1.07 |
Table 15. Summary of Coagulation Values
F0 Animals – Males, Day: 137 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.55 0.20 10 - | 13.51 1.35 10 - | 2.278 0.177 10 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.43 0.15 10 0.99 | 12.52 1.72 10 0.93 | 2.282 0.122 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.30 ** 0.14 10 0.98 | 12.35 1.14 10 0.91 | 2.286 0.185 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.26 ** 0.12 10 0.97 | 11.77 1.51 10 0.87 | 2.454 * 0.114 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 16. Summary of Coagulation Values
F0 Animals – Females, Day: 115 Relative to Start Date
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.19 0.21 9 - | 11.17 1.22 9 - | 1.713 0.292 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.48 0.34 10 1.03 | 12.17 1.55 10 1.09 | 1.612 0.157 10 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.27 0.32 9 1.01 | 11.82 1.49 9 1.06 | 1.743 0.316 9 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.39 0.73 10 1.02 | 11.16 2.64 10 1.00 | 1.682 0.425 10 0.98 |
[G] - Anova & Dunnett
Table 17. Summary of Coagulation Values
F1 Animals - Cohort 1A, Day: 70 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.50 0.14 9 - | 12.27 1.80 9 - | 2.580 0.172 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.40 0.11 10 0.99 | 12.79 2.10 10 1.04 | 2.516 0.141 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.34 * 0.14 10 0.98 | 12.09 2.30 10 0.99 | 2.655 0.219 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.33 * 0.05 9 0.98 | 11.51 2.25 9 0.94 | 2.707 0.250 9 1.05 |
[G] - Anova & Dunnett: * = p ≤ 0.05
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.31 0.26 9 - | 12.21 1.52 8 - | 2.116 0.251 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.17 0.17 9 0.99 | 12.33 1.57 9 1.01 | 2.128 0.172 9 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.15 0.14 10 0.98 | 11.65 0.99 10 0.95 | 2.186 0.194 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.02 ** 0.17 10 0.97 | 11.71 1.65 10 0.96 | 2.182 0.099 10 1.03 |
[G] - Anova & Dunnett: ** = p ≤ 0.01
Table 18. Summary of Hematology Values
F0 Animals – Males Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G1] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 4.651 0.478 10 - | 0.961 0.257 10 - | 3.480 0.289 10 - | 0.065 0.014 10 - | 0.116 0.046 10 - | 0.008 0.004 10 - | 0.019 0.006 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 4.340 0.805 10 0.93 | 0.769 0.202 10 0.80 | 3.390 0.784 10 0.97 | 0.062 0.018 10 0.95 | 0.096 0.024 10 0.83 | 0.006 0.005 10 0.75 | 0.016 0.008 10 0.84 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.469 0.783 10 0.96 | 0.687 * 0.169 10 0.71 | 3.605 0.697 10 1.04 | 0.057 0.021 10 0.88 | 0.089 0.023 10 0.77 | 0.010 0.000 10 1.25 | 0.019 0.007 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.192 0.960 10 1.12 | 0.854 0.199 10 0.89 | 4.126 0.908 10 1.19 | 0.079 0.028 10 1.22 | 0.092 0.040 10 0.79 | 0.009 0.006 10 1.13 | 0.027 0.013 10 1.42 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 8.813 0.449 10 - | 153.6 4.7 10 . | 0.4464 0.0164 10 - | 50.68 1.37 10 - | 17.43 0.57 10 - | 343.9 7.0 10 - | 11.88 0.31 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.630 0.303 10 0.98 | 150.7 6.5 10 0.98 | 0.4441 0.0155 10 0.99 | 51.52 2.26 10 1.02 | 17.46 1.03 10 1.00 | 338.9 7.5 10 0.99 | 12.00 0.48 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.572 0.317 10 0.97 | 149.6 5.2 10 0.97 | 0.4388 0.0129 10 0.98 | 51.25 1.89 10 1.01 | 17.47 0.71 10 1.00 | 341.1 6.6 10 0.99 | 12.01 0.52 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.573 0.220 10 0.97 | 147.4 5.4 10 0.96 | 0.4364 0.0139 10 0.98 | 50.92 1.39 10 1.00 | 17.17 0.55 10 0.99 | 337.5 4.5 10 0.98 | 12.16 0.48 10 1.02 |
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 781.6 146.2 10 - | 155.83 19.62 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 765.3 76.5 10 0.98 | 142.51 41.33 10 0.91 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 760.4 92.6 10 0.97 | 160.37 18.17 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 883.4 160.3 10 1.13 | 171.47 21.32 10 1.10 |
Table 19. Summary of Hematology Values
F0 Animals – Females Day: 115 Relative to Start Date
Sex: Female |
| Reporting Hematology | ||||||
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.168 1.072 8 - | 1.669 0.559 8 - | 3.265 0.790 8 - | 0.109 0.031 8 - | 0.095 0.038 8 - | 0.011 0.006 8 - | 0.023 0.009 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.529 0.603 10 1.07 | 1.737 0.408 10 1.04 | 3.564 0.620 10 1.09 | 0.113 0.025 10 1.04 | 0.077 0.039 10 0.81 | 0.012 0.008 10 1.07 | 0.025 0.010 10 1.11 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 5.866 1.252 10 1.14 | 1.903 0.424 10 1.14 | 3.694 0.894 10 1.13 | 0.143 0.060 10 1.31 | 0.091 0.038 10 0.96 | 0.013 0.007 10 1.16 | 0.026 0.013 10 1.16 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.570 0.802 10 1.08 | 1.762 0.315 10 1.06 | 3.555 0.672 10 1.09 | 0.144 0.051 10 1.32 | 0.073 0.013 10 0.77 | 0.008 0.004 10 0.71 | 0.028 0.011 10 1.24 |
Sex: Female |
| Reporting Hematology | ||||||
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.958 0.449 8 - | 157.3 3.4 8 . | 0.4679 0.0124 8 - | 58.88 2.37 8 - | 19.80 0.92 8 - | 336.4 5.2 8 - | 12.00 1.06 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.678 0.380 10 0.96 | 153.3 5.2 10 0.97 | 0.4515 0.0208 10 0.97 | 58.86 1.56 10 1.00 | 19.99 0.77 10 1.01 | 339.8 8.3 10 1.01 | 11.84 0.79 10 0.99 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.466 0.606 9 0.94 | 152.7 5.1 10 0.97 | 0.4468 0.0194 9 0.95 | 59.99 2.69 9 1.02 | 20.48 1.21 9 1.03 | 341.6 7.1 9 1.02 | 11.82 0.97 10 0.99 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.528 0.566 10 0.95 | 151.3 8.8 10 0.96 | 0.4428 0.0316 10 0.95 | 58.85 1.83 10 1.00 | 20.10 1.07 10 1.02 | 341.5 10.8 10 1.02 | 12.17 1.05 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 894.6 119.2 8 - | 177.99 20.44 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 794.3 132.8 10 0.89 | 178.18 31.61 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 872.4 121.5 9 0.98 | 161.09 22.77 10 0.91 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 833.4 177.2 10 0.93 | 173.25 30.61 10 0.97 |
Table 20. Summary of Hematology Values
F1 Animals – Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 6.989 1.038 10 - | 0.930 0.183 10 - | 5.762 0.940 10 - | 0.121 0.043 10 - | 0.108 0.036 10 - | 0.008 0.009 10 - | 0.059 0.018 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.203 0.765 10 1.03 | 0.970 0.322 10 1.04 | 5.976 0.743 10 1.04 | 0.099 0.031 10 0.82 | 0.091 0.031 10 0.84 | 0.011 0.006 10 1.38 | 0.054 0.023 10 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.256 0.780 10 1.04 | 0.993 0.193 10 1.07 | 6.050 0.666 10 1.05 | 0.086 0.030 10 0.71 | 0.074 0.023 10 0.69 | 0.007 0.005 10 0.88 | 0.047 0.009 10 0.80 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.090 * 0.978 10 1.16 | 1.081 0.253 10 1.16 | 6.736 1.071 10 1.17 | 0.115 0.030 10 0.95 | 0.080 0.032 10 0.74 | 0.011 0.006 10 1.38 | 0.064 0.023 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Hematology |
|
|
| |
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 9.078 0.461 10 - | 163.2 4.9 10 . | 0.4747 0.0207 10 - | 52.33 1.52 10 - | 17.99 0.90 10 - | 343.7 10.9 10 - | 11.47 0.49 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.551 * 0.488 10 0.94 | 156.9 * 5.4 10 0.96 | 0.4471 ** 0.0181 10 0.94 | 52.33 2.09 10 1.00 | 18.37 0.87 10 1.02 | 350.8 6.2 10 1.02 | 11.59 0.52 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.522 ** 0.298 10 0.94 | 157.1 * 4.1 10 0.96 | 0.4519 * 0.0171 10 0.95 | 53.06 1.54 10 1.01 | 18.44 0.54 10 1.03 | 347.7 7.0 10 1.01 | 11.63 0.51 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.340 ** 0.285 10 0.92 | 154.7 ** 4.4 10 0.95 | 0.4456 ** 0.0166 10 0.94 | 53.45 1.51 10 1.02 | 18.56 0.67 10 1.03 | 347.4 7.6 10 1.01 | 11.84 0.32 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 763.2 131.0 10 - | 193.85 15.23 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 876.8 89.5 10 1.15 | 171.56 23.85 10 0.89 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 899.7 76.1 10 1.18 | 198.49 23.89 10 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 823.3 144.5 10 1.08 | 208.78 30.71 10 1.08 |
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G1] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G1] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.104 1.013 9 - | 0.587 0.094 9 - | 4.321 0.997 9 - | 0.070 0.018 9 - | 0.081 0.019 9 - | 0.003 0.005 9 - | 0.038 0.010 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.207 1.111 9 1.02 | 0.671 0.142 9 1.14 | 4.371 1.058 9 1.01 | 0.053 0.017 9 0.76 | 0.074 0.028 9 0.92 | 0.004 0.005 9 1.33 | 0.029 0.014 9 0.76 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.656 0.870 10 0.91 | 0.708 0.238 10 1.21 | 3.780 0.791 10 0.87 | 0.060 0.021 10 0.86 | 0.067 0.020 10 0.83 | 0.003 0.005 10 0.90 | 0.034 0.008 10 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.515 1.387 10 1.08 | 0.681 0.323 10 1.16 | 4.640 1.417 10 1.07 | 0.076 0.039 10 1.09 | 0.079 0.031 10 0.97 | 0.004 0.005 10 1.20 | 0.037 0.014 10 0.98 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.842 0.436 9 - | 151.6 5.3 9 . | 0.4248 0.0164 9 - | 54.23 1.82 9 - | 19.38 0.95 9 - | 356.8 8.1 9 - | 10.70 0.38 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.868 0.225 9 1.00 | 148.8 6.5 9 0.98 | 0.4244 0.0173 9 1.00 | 53.91 1.31 9 0.99 | 18.89 0.64 9 0.97 | 350.7 8.5 9 0.98 | 10.91 0.36 9 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.867 0.362 10 1.00 | 149.7 5.4 10 0.99 | 0.4244 0.0143 10 1.00 | 54.01 2.25 10 1.00 | 19.06 0.93 10 0.98 | 352.6 4.3 10 0.99 | 10.68 0.35 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.744 0.368 10 0.99 | 144.3 5.5 10 0.95 | 0.4116 0.0156 10 0.97 | 53.20 1.94 10 0.98 | 18.64 0.75 10 0.96 | 350.5 6.8 10 0.98 | 10.82 0.33 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 833.0 141.8 9 - | 183.07 34.30 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 801.3 145.2 8 0.96 | 178.02 22.41 9 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 796.3 97.2 9 0.96 | 185.18 28.61 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 910.6 58.3 10 1.09 | 177.80 38.40 10 0.97 |
Table 21. Summary of Reproductive Performance
F0 Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 25 25 25 5.3 5.1 25 - 19 76.0 6 24.0 100.0 25/25 | 25 25 25 3.2 2.5 25 -39.4 24 96.0 1 4.0 100.0 25/25 | 25 25 25 2.7 2.6 25 -49.2 24 96.0 1 4.0 100.0 25/25 | 25 24 24 3.0 2.7 24 -43.2 23 95.8 1 4.2 100.0 24/24 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 22. Summary of Reproductive Performance
F1 Animals - Cohort 1B Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 20 20 20 2.7 1.0 20 - 20 100.0 0 0.0 100.0 20/20 | 20 20 20 3.9 3.7 20 45.3 18 90.0 2 10.0 100.0 20/20 | 20 20 20 3.5 2.7 20 32.1 19 95.0 1 5.0 100.0 20/20 | 20 20 20 2.5 1.2 20 -7.5 20 100.0 0 0.0 100.0 20/20 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 23. Mating Performance and Fertility
F0 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 6(1)* | 4 | 8 | 7(1) |
2 | 2 | 5 | 6 | 5 |
3 | 6 | 8 | 7 | 3 |
4 | 5 | 7 | 3 | 8 |
No clear indication of mating | 6(1) | 1 | 1 | 1 |
Mean number of nights to positive mating sign | 2.5 | 2.8 | 2.2 | 2.5 |
Number passing one estrus | 0 | 0 | 0 | 0 |
Number of males paired | 25 | 25 | 25 | 24 |
Number of siring males | 24 | 25 | 25 | 23 |
Male Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Male Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Male Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
Female Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Female Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Female Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
*Includes Animal 1524 that had 2 implant sites (no live or dead pups)
Table 24. Mating Performance and Fertility
F1 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 3 | 5 | 2 | 7 |
2 | 5 | 2 | 4 | 2 |
3 | 9 | 5(2) | 8 | 6 |
4 | 2 | 5 | 4 | 5 |
5 | 1(1) | 1 | 0 | 0 |
6 | 0 | 0 | 1 | 0 |
No clear indication of mating | 0 | 2(2) | 1(1) | 0 |
Mean number of nights to positive mating sign | 2.7 | 2.7 | 2.9 | 2.5 |
Number passing one estrus | 0 | 1 | 1 | 0 |
Number of males paired | 20 | 20 | 20 | 20 |
Number of siring males | 19 | 16 | 19 | 20 |
Male Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Male Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Male Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Female Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Female Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Female Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Table 25. Summary of Duration of Gestation and Overall Litter Performance
F0 Animals
F0 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 24 | 25 | 25 | 23 |
Duration of Gestation (Days) 21 |
1 |
1 |
0 |
0 |
22 | 17 | 17 | 23 | 20 |
23 | 0 | 6 | 1 | 2 |
No clear indication of mating | 6 | 1 | 1 | 1 |
Mean Duration | 21.9 | 22.2 | 22.0 | 22.1 |
Number of females producing a live litter | 23 | 25 | 25 | 23 |
Gestation index as % | 96 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.0 ± 3.4 | 12.5 ± 2.4 | 12.6 ± 1.9 | 12.9 ± 1.6 |
Mean total number of pups born per litter ± standard deviation | 11.0 ± 3.0 | 11.2 ± 2.3 | 11.0 ± 2.5 | 11.0 ± 3.3 |
Mean number of live pups per litter ± standard deviation: Lactation Day 1 |
11.0 ± 3.0 |
11.1 ± 2.2 |
10.9 ± 2.6 |
11.1 ± 2.6 |
Lactation Day 4 | 10.9 ± 3.1 | 11.2 ± 2.2 | 10.9 ± 2.6 | 11.0 ± 2.7 |
Lactation Day 4a | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 7 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8± 0.7 | 7.7 ± 0.9 |
Lactation Day 14 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 21 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Number of males on Lactation Day 1 (%) | 120 (47) | 135 (49) | 145 (53) | 116 (47) |
Number of females on Lactation Day 1 (%) | 134 (53) | 143 (51) | 127 (47) | 129 (53) |
a After cull of pups on PND 4 to standardise the size of the litter
Table 26. Summary of Duration of Gestation and Overall Litter Performance
F1 Animals
F1 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 19 | 16 | 19 | 20 |
Duration of Gestation (Days) 21 |
1 |
3 |
1 |
1 |
22 | 15 | 9 | 17 | 18 |
23 | 3 | 4 | 1 | 1 |
Mean Duration | 22.1 | 22.1 | 22.0 | 22.0 |
Number of females producing a live litter | 19 | 16 | 19 | 20 |
Gestation index as % | 100 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.6 ± 1.7 | 12.3 ± 2.3 | 13.0 ± 1.6 | 12.5 ± 2.3 |
Mean total number of pups born per litter ± standard deviation* | 11.6 ± 1.9 | 11.1 ± 2.8 | 10.5 ± 2.8 | 11.3 ± 2.2 |
Mean number of live pups per litter ± standard deviation*: Lactation Day 1 |
11.3 ± 2.2 |
11.1 ± 3.0 |
10.4 ± 2.7 |
11.1 ± 2.1 |
Lactation Day 4 | 11.3 ± 2.2 | 11.0 ± 2.9 | 10.3 ± 2.8 | 11.0 ± 2.1 |
Lactation Day 4a | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 7 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 14 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 21 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.8 ± 0.5 |
Number of males on Lactation Day 1 (%) | 96 (47) | 96 (54) | 93 (49) | 100 (45) |
Number of females on Lactation Day 1 (%) | 108 (53) | 81 (46) | 96 (51) | 121 (55) |
a After cull of pups on PND 4 to standardise the size of the litter
* Excludes Animal 1628 that had a total litter loss
Table 27. Summary of Urinalysis Values
F0 Animals - Males
Day: 131 Relative to Start Date
| Sex: Male | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G1] | |||
| 0 mg/kg/day | Mean | 4.32 | 1.0152 | 7.5 |
| Group 1 | SD | 2.94 | 0.0089 | 0.94 |
| N | 10 | 10 - | 10 - | |
| - | ||||
| 100 mg/kg/day | Mean | 5.64 | 1.0135 | 8 |
| Group 2 | SD | 2.99 | 0.0052 | 0.62 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.31 | 1 | 1.07 | |
| 300 mg/kg/day | Mean | 4.22 | 1.0215 | 8.15 |
| Group 3 | SD | 2.11 | 0.0086 | 0.63 |
| N | 10 | 10 | 10 | |
| tCtrl | 0.98 | 1.01 | 1.09 | |
| 900 mg/kg/day | Mean | 5.25 | 1.0300 ** | 7.35 0.71 |
| Group 4 | SD | 2.39 | 0.0133 | 10 |
| N | 10 | 10 | 0.98 | |
| tCtrl | 1.22 | 1.01 |
Table 28. Summary of Urinalysis Values
F0 Animals - Females
Day: 108 Relative to Start Date
| Sex: Female | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G1] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 4.85 | 1.0188 | 8.6 |
| Group 1 | SD | 2.33 | 0.0052 | 0.32 |
| N | 10 - | 10 - | 10 - | |
| 100 mg/kg/day | Mean | 5.23 | 1.0192 | 8.45 |
| Group 2 | SD | 2.96 | 0.0069 | 0.16 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.08 | 1 | 0.98 | |
| 300 mg/kg/day | Mean | 6.01 | 1.0199 | 8.2 |
| Group 3 | SD | 1.68 | 0.0039 | 0.42 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.24 | 1 | 0.95 | |
| 900 mg/kg/day | Mean | 7.05 2.58 | 1.0218 0.0051 | 8.05 * |
| Group 4 | SD | 10 | 10 | 0.72 |
| N | 1.45 | 1 | 10 | |
| tCtrl | 0.94 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05
Table 29. Summary of Urinalysis Values
F1 Animals - Cohort 1A
Day: 60 Relative to Start Date
| Sex: Male | Reporting urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 2.67 | 1.0182 | 8.11 |
| Group 1 | SD | 1.87 | 0.0078 | 0.6 |
| N | 10 - | 10 - | 9 - | |
| 100 mg/kg/day | Mean | 3.07 | 1.0168 | 7.9 |
| Group 2 | SD | 0.95 | 0.0123 | 0.61 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.15 | 1 | 0.97 | |
| 300 mg/kg/day | Mean | 1.96 | 1.0302 * | 7.11 ** 0.70 |
| Group 3 | SD | 1.25 | 0.0118 | 9 |
| N | 10 | 10 | 0.88 | |
| tCtrl | 0.73 | 1.01 | ||
| 900 mg/kg/day | Mean | 2.94 1.43 | 1.0346 ** | 6.25 ** |
| Group 4 | SD | 10 | 0.0091 | 0.42 |
| N | 1.1 | 10 | 10 | |
| tCtrl | 1.02 | 0.77 |
Day: 60 Relative to Start Date
Sex: Female |
|
| Reporting Urinalysi | s |
VOLUME (mL) [G] | SPECIFIC GRAVITY [G] | URINE pH [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.88 1.25 10 - | 1.0226 0.0117 10 - | 7.00 0.94 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 3.20 2.88 10 1.70 | 1.0161 0.0086 10 0.99 | 7.15 0.91 10 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.33 1.08 10 0.71 | 1.0333 0.0118 10 1.01 | 6.61 0.55 9 0.94 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.61 1.55 9 1.39 | 1.0267 0.0087 9 1.00 | 6.44 0.68 9 0.92 |
[G] - Anova & Dunnett
Table 1 - Summary of Body Weights (g)
F0 Males
| Sex: Male | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 124.9 | 176.6 | 225.1 | 268.9 | 303.1 | 333 | 355.5 |
| Group 1 | SD | 16.6 | 18.5 | 21 | 23.8 | 24.7 | 26.3 | 27.3 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 123.8 | 174.3 | 225.7 | 270.7 | 305.6 | 333.2 | 355.7 |
| Group 2 | SD | 16.3 | 18.8 | 23.4 | 25.2 | 26.4 | 28.5 | 30.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.9 | -1.3 | 0.3 | 0.7 | 0.8 | 0.1 | 0.1 | |
| 300 mg/kg/day | Mean | 121 | 171.9 | 222.1 | 268.3 | 302.8 | 332.4 | 355.9 |
| Group 3 | SD | 15.1 | 18.9 | 21 | 23.1 | 22.1 | 24.6 | 26.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -3.1 | -2.7 | -1.3 | -0.2 | -0.1 | -0.2 | 0.1 | |
| 900 mg/kg/day | Mean | 124.9 | 178.2 | 230.5 | 277.9 | 310.1 | 334.8 | 352.2 |
| Group 4 | SD | 11.6 | 14.2 | 18.9 | 21.9 | 24.1 | 25.7 | 28.3 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0 | 0.9 | 2.4 | 3.4 | 2.3 | 0.5 | -0.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 43 | 50 | 57 | 64 | 71 | 78 | 85 | ||
| 0 mg/kg/day | Mean | 374.2 | 389 | 406.9 | 417.8 | 427.7 | 428 | 435.6 |
| Group 1 | SD | 30.2 | 31.6 | 33.7 | 35 | 34.9 | 33.2 | 34.4 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 374.4 | 392.8 | 408 | 417.3 | 423.9 | 427.8 | 437.4 |
| Group 2 | SD | 32.8 | 34.9 | 36.1 | 35.5 | 36.7 | 37.4 | 37.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0.1 | 1 | 0.3 | -0.1 | -0.9 | -0.1 | 0.4 | |
| 300 mg/kg/day | Mean | 373.7 | 390.5 | 404.8 | 415.6 | 424 | 426.6 | 436.5 |
| Group 3 | SD | 29.8 | 32.2 | 33.2 | 33.8 | 36.9 | 36.4 | 38.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | 0.4 | -0.5 | -0.5 | -0.9 | -0.3 | 0.2 | |
| 900 mg/kg/day | Mean | 365.9 | 378.8 | 393.4 | 403.8 | 411.4 | 415.7 | 423.1 |
| Group 4 | SD | 29.7 | 32 | 33.1 | 34.7 | 35.5 | 34.6 | 36.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -2.2 | -2.6 | -3.3 | -3.3 | -3.8 | -2.9 | -2.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 92 | 99 | 106 | 113 | 120 | 127 | 134 | ||
| 0 mg/kg/day | Mean | 441.8 | 453.6 | 466.3 | 472.8 | 483.5 | 489.2 | 496.7 |
| Group 1 | SD | 35.9 | 36.1 | 37.8 | 37.8 | 38.9 | 39 | 41.7 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 441.6 | 450.2 | 462.2 | 470.4 | 478 | 487.7 | 492.6 |
| Group 2 | SD | 37.1 | 37.6 | 39.7 | 40.7 | 41.1 | 43.8 | 45.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | -0.7 | -0.9 | -0.5 | -1.1 | -0.3 | -0.8 | |
| 300 mg/kg/day | Mean | 446.7 | 452.6 | 463.6 | 475.6 | 483.9 | 492.7 | 496.6 |
| Group 3 | SD | 40.7 | 42.3 | 44.7 | 47 | 49.7 | 50.9 | 53.1 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 1.1 | -0.2 | -0.6 | 0.6 | 0.1 | 0.7 | 0 | |
| 900 mg/kg/day | Mean | 434.1 | 442.2 | 449.5 | 458.4 | 464.4 | 475.5 | 478.6 |
| Group 4 | SD | 38 | 39.8 | 40.3 | 40.7 | 40.7 | 42.5 | 44.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -1.7 | -2.5 | -3.6 | -3 | -3.9 | -2.8 | -3.6 | |
| Sex: Male | Day(s) Relative to Start Date | ||||
| 137 | 138 | 141 | 142 | ||
| 0 mg/kg/day | Mean | 507.9 | 501.8 | 493.1 | 473.4 |
| Group 1 | SD | 58.2 | 33.5 | 36.1 | 43.3 |
| N | 8 - | 6 - | 11 - | 5 - | |
| 100 mg/kg/day | Mean | 488 | 512.6 | 490.8 | 477.7 |
| Group 2 | SD | 46.5 | 39.6 | 50.5 | 63.9 |
| N | 5 | 7 | 13 | 7 | |
| %Diff | -3.9 | 2.1 | -0.5 | 0.9 | |
| 300 mg/kg/day | Mean | 458 | 534.4 | 491 | 494.8 |
| Group 3 | SD | 26.4 | 57.8 | 46.4 | 52.9 |
| N | 4 | 8 | 13 | 8 | |
| %Diff | -9.8 | 6.5 | -0.4 | 4.5 | |
| 900 mg/kg/day | Mean | 473.8 | 494.5 | 480.2 | 471.2 |
| Group 4 | SD | 41.8 | 27.5 | 54.1 | 65.1 |
| N | 8 | 4 | 13 | 5 | |
| %Diff | -6.7 | -1.5 | -2.6 | -0.5 | |
Table 2 - Summary of Body Weights (g)
F0 Females - Prior to mating
| Sex: Female | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 72.9 | 110.5 | 136.4 | 155.4 | 173.6 | 188.9 | 197.4 |
| Group 1 | SD | 13.5 | 13.9 | 14 | 13.8 | 13.3 | 13.9 | 14.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 76 | 114.9 | 140.8 | 162 | 181.1 | 195.7 | 206.3 |
| Group 2 | SD | 10.2 | 11.1 | 11.6 | 13.5 | 14.4 | 14.5 | 16.5 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 3.9 | 3.3 | 4.3 | 4.3 | 3.6 | 4.5 | |
| 300 mg/kg/day | Mean | 74.9 | 114.2 | 142.2 | 162.9 | 183.5 | 198.8 | 209.1* |
| Group 3 | SD | 12.4 | 11.1 | 10.6 | 12.3 | 12.8 | 14.7 | 15.6 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 2.8 | 3.4 | 4.3 | 4.9 | 5.7 | 5.2 | 5.9 | |
| 900 mg/kg/day | Mean | 77.2 | 116.4 | 147.2* | 171.3** | 191.6** | 208.2** | 214.9** |
| Group 4 | SD | 14.1 | 14.3 | 15.3 | 17.2 | 17.4 | 17.9 | 18.2 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 5.9 | 5.3 | 7.9 | 10.3 | 10.3 | 10.2 | 8.9 |
| Sex: Female | Day(s) Relative to Start Date | |||||
| 43 | 50 | 57 | 64 | 71 | ||
| 0 mg/kg/day | Mean | 207.3 | 214.6 | 222.2 | 226.2 | 230.5 |
| Group 1 | SD | 14.9 | 13.1 | 14.8 | 14.4 | 15.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 216 | 225.6 | 229.6 | 232.8 | 240.8 |
| Group 2 | SD | 17.1 | 18.4 | 19 | 20.5 | 20.5 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 5.2 | 3.3 | 2.9 | 4.5 | |
| 300 mg/kg/day | Mean | 217.2 | 227.9* | 234.7* | 238.4 | 245.9* |
| Group 3 | SD | 16.5 | 18.6 | 19.4 | 18.9 | 19 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.8 | 6.2 | 5.6 | 5.4 | 6.7 | |
| 900 mg/kg/day | Mean | 227.0** | 233.0** | 242.0** | 244.5** | 250.5** |
| Group 4 | SD | 17.2 | 17.5 | 18.6 | 19 | 19.7 |
| N | 24 | 24 | 24 | 24 | 24 | |
| %Diff | 9.5 | 8.6 | 8.9 | 8.1 | 8.7 |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 3 - Summary of Body Weights (g)
F0 Females - Gestation
| Sex: Female | Day(s) Relative to Mating (Litter: A) | ||||
| 0 | 7 | 14 | 20 | ||
| 0 mg/kg/day | Mean | 222.8 | 246.1 | 269.9 | 325.9 |
| Group 1 | SD | 14.5 | 13.5 | 16.3 | 24 |
| N | 18 - | 18 - | 18 - | 18 - | |
| 100 mg/kg/day | Mean | 237.2* | 259.5 | 282.3 | 338.5 |
| Group 2 | SD | 20.1 | 20.5 | 22.9 | 31.2 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 6.5 | 5.5 | 4.6 | 3.9 | |
| 300 mg/kg/day | Mean | 242.1** | 262.4* | 284.2 | 343.6 |
| Group 3 | SD | 20.5 | 22.4 | 24.6 | 28.1 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 8.6 | 6.6 | 5.3 | 5.4 | |
| 900 mg/kg/day | Mean | 243.7** | 264.5* | 291.1* | 352.5* |
| Group 4 | SD | 17.2 | 21.1 | 23.4 | 34.4 |
| N | 22 | 22 | 22 | 22 | |
| %Diff | 9.4 | 7.5 | 7.9 | 8.2 | |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 4 - Summary of Body Weights (g)
F0 Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | ||
| Sex | 7 | ||||||
| 1F | Mean | 261.7 | 274 | 275.1 | 282.8 | 280.1 | |
| SD | 19.1 | 20.8 | 19.3 | 21.2 | 16.6 | ||
| N | 23 | 22 | 23 | 23 | 23 | ||
| Mean | 268.2 | 280.9 | 283.3 | 290.9 | 288 | ||
| 2F | SD | 22.1 | 22.6 | 23 | 23.6 | 21.7 | |
| N | 23 | 24 | 24 | 24 | 24 | ||
| %Diff G1 | 2.5 | 2.5 | 3 | 2.9 | 2.8 | ||
| Mean | 270.8 | 282.7 | 285.6 | 294.8 | 291 | ||
| SD | 25.7 | 25 | 24.4 | 23.4 | 19.9 | ||
| 3F | N | 25 | 25 | 25 | 25 | 25 | |
| %Diff G1 | 3.5 | 3.2 | 3.8 | 4.2 | 3.9 | ||
| Mean | 271.6 | 286.4 | 292.2 | 302.6a | 300.1b | ||
| 4F | SD | 23.4 | 24 | 24.8 | 25.7 | 25.2 | |
| N | 22 | 22 | 22 | 22 | 22 | ||
| %Diff G1 | 3.8 | 4.5 | 6.2 | 7 | 7.1 | ||
Significantly different from control group 1 value :a=p≤0.05,b=p≤0.01 (Dunnett)
Table 5 - Summary of Body Weights (g)
F1 Animals - Cohort 1A
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
| 0 mg/kg/day | Mean | 56 | - | 71.8 | 119.9 | 172.1 | 219.5 | 269.6 |
| Group 1 | SD | 5.1 | - | 8.8 | 12.2 | 15.8 | 19.7 | 23.4 |
| N | 20 - | - | 20 - | 20 - | 20 - | 20 - | 20 - | |
| - | ||||||||
| 100 mg/kg/day | Mean | 56.3 | 61.0n | 70.8 | 116.4 | 166.9 | 213.7 | 261.2 |
| Group 2 | SD | 6.6 | 6.6 | 9 | 13.6 | 16.5 | 20.5 | 26.4 |
| N | 19 | 3 - | 18 | 20 | 20 | 20 | 20 | |
| %Diff | 0.7 | -1.4 | -2.9 | -3 | -2.6 | -3.1 | ||
| 300 mg/kg/day | Mean | 57.9 | 76.0n | 76.8 | 126.3 | 179.8 | 227 | 276.8 |
| Group 3 | SD | 4.9 | - | 9.3 | 14 | 16.9 | 19.6 | 24.9 |
| N | 20 | 1 - | 19 | 20 | 20 | 20 | 20 | |
| %Diff | 3.5 | 6.9 | 5.4 | 4.5 | 3.4 | 2.7 | ||
| 900 mg/kg/day | Mean | 54.7 | 59.0n 4.3 | 70.6 | 115.8 | 167.5 | 212.9 | 258.5 |
| Group 4 | SD | 5.5 | 6 - | 8.2 | 12.5 | 16.5 | 21.7 | 25.5 |
| N | 15 | 19 | 20 | 20 | 20 | 20 | ||
| %Diff | -2.2 | -1.6 | -3.4 | -2.7 | -3 | -4.1 |
[G] - Anova & Dunnett [I] - n - Inappropriate for statistics
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 41 | 48 | 55 | 62 | 68 | 69 | 70 | ||
| 0 mg/kg/day | Mean | 309.2 | 343.6 | 368.2 | 390.7 | 408.1 | 394 | 405.5 |
| Group 1 | SD | 26.7 | 31.5 | 33 | 37 | 40.1 | 42.2 | 40.7 |
| N | 20 - | 20 - | 20 - | 20 - | 14 - | 6 - | 20 - | |
| 100 mg/kg/day | Mean | 300.1 | 335.3 | 360.5 | 382.8 | 393 | 408.2 | 401.6 |
| Group 2 | SD | 29.4 | 32.6 | 35.7 | 36.5 | 35.3 | 42.7 | 34.3 |
| N | 20 | 20 | 20 | 20 | 16 | 6 | 16 | |
| %Diff | -2.9 | -2.4 | -2.1 | -2 | -3.7 | 3.6 | -1 | |
| 300 mg/kg/day | Mean | 317.3 | 349.4 | 376.1 | 396.8 | 411.1 | 445.9 | 411.1 |
| Group 3 | SD | 28 | 33 | 37.6 | 38.7 | 46.2 | 36.6 | 36.1 |
| N | 20 | 20 | 20 | 20 | 14 | 7 | 19 | |
| %Diff | 2.6 | 1.7 | 2.1 | 1.6 | 0.8 | 13.2 | 1.4 | |
| 900 mg/kg/day | Mean | 299.2 | 325.5 | 346.4 | 363.5 | 386.8 | 381.7 | 382.7 |
| Group 4 | SD | 29 | 32.1 | 37.5 | 40 | 40.6 | 48.2 | 38.9 |
| N | 20 | 20 | 20 | 20 | 14 | 11 | 15 | |
| %Diff | -3.2 | -5.3 | -5.9 | -7 | -5.2 | -3.1 | -5.6 |
Sex: Male |
| Day(s) Relative to Start Date |
71 | ||
0 mg/kg/day
Group 1 | Mean SD N
| - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 447.0n 69.3 2 - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | - - - - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | - - - - |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 4.9 20 - | - - - - | 66.0 7.8 20 - | 103.8 9.7 20 - | 136.5 10.7 20 - | 157.2 10.2 20 - | 176.5 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 55.3 4.9 19 5.3 | 58.0n 3.0 3 - | 68.9 5.7 18 4.5 | 106.1 9.4 20 2.2 | 138.3 11.9 20 1.3 | 159.7 14.1 20 1.6 | 178.8 16.8 20 1.3 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.0 5.0 21 4.7 | 71.0n - 1 - | 69.4 8.9 19 5.2 | 105.5 11.3 20 1.7 | 138.6 11.6 20 1.5 | 160.2 12.3 20 1.9 | 179.9 13.8 20 1.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 52.1 4.8 15 -0.8 | 56.3n 6.0 7 - | 66.1 8.2 18 0.2 | 103.2 10.6 20 -0.6 | 138.6 12.8 20 1.5 | 159.9 15.9 20 1.7 | 182.0 15.5 20 3.1 |
[G] - Anova & Dunnett
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 [G] | 48 [G] | 55 [G] | 62 [G] | 68 [G] | 69 [G] | 70 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.0 12.1 20 - | 203.2 14.4 20 - | 212.2 15.5 20 - | 221.0 18.8 20 - | 231.6 18.8 13 - | 228.1 18.9 7 - | 245.3n 11.0 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 192.4 18.5 20 1.8 | 205.7 17.3 20 1.2 | 214.1 18.0 20 0.9 | 224.2 20.5 20 1.4 | 235.2 22.6 17 1.6 | 216.7 6.8 3 -5.0 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 194.4 13.6 20 2.8 | 206.5 14.6 20 1.6 | 216.4 16.9 20 2.0 | 226.1 17.7 20 2.3 | 231.5 16.3 14 0.0 | 242.3 12.6 6 6.2 | 220.0n - 1 -10.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 201.6 17.1 20 6.7 | 209.2 17.6 20 3.0 | 218.0 17.7 20 2.7 | 226.5 18.3 20 2.5 | 237.5 19.1 15 2.6 | 235.6 26.0 5 3.3 | - - - - |
[G] - Anova & Dunnett
Sex: Female |
| Day(s) Relative to Start Date |
| ||
71 [G] | 72 [I] | 73 [G] | 74 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 226.9 16.6 8 - | - - - - | 228.6 21.1 5 - | 228.8 17.9 4 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 228.3 29.4 8 0.6 | 223.7n 12.2 3 - | 233.8 11.4 5 2.3 | 233.8 25.4 4 2.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 233.1 15.5 9 2.7 | 268.0n - 1 - | 225.0n 11.3 2 -1.6 | 237.7 20.5 7 3.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 235.0 23.9 10 3.6 | 247.0n 11.3 2 - | 232.8 19.2 6 1.9 | 235.0n 25.5 2 2.7 |
[G] - Anova & Dunnett: n - Inappropriate for statistics
Table 7 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Males
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 5.4 20 - | - - - - | 70.5 8.5 20 - | 116.5 12.0 20 - | 167.7 13.3 20 - | 212.9 17.3 20 - | 261.7 20.9 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 57.4 5.2 19 5.1 | 60.0n 8.5 3 - | 72.9 8.7 18 3.5 | 117.5 14.9 20 0.8 | 166.8 20.5 20 -0.6 | 211.2 25.9 20 -0.8 | 258.5 29.7 20 -1.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 56.8 5.7 20 4.0 | 70.0n - 1 - | 74.9 10.8 19 6.2 | 124.1 15.9 20 6.5 | 175.7 18.3 20 4.7 | 222.6 21.4 20 4.6 | 271.2 25.8 20 3.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 54.5 4.5 15 -0.1 | 59.3n 5.9 7 - | 69.9 8.7 18 -0.8 | 115.4 10.8 20 -0.9 | 166.8 12.3 20 -0.5 | 212.2 15.2 20 -0.3 | 258.6 18.7 20 -1.2 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 300.5 25.6 20 - | 334.6 30.5 20 - | 358.4 34.8 20 - | 381.5 38.4 20 - | 395.5 41.5 20 - | 410.3 44.0 20 - | 421.2 46.7 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 297.5 32.5 20 -1.0 | 330.9 34.9 20 -1.1 | 357.3 37.9 20 -0.3 | 379.6 39.8 20 -0.5 | 399.5 39.4 20 1.0 | 414.5 41.7 20 1.0 | 429.9 42.0 20 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 310.2 26.4 20 3.2 | 342.6 29.7 20 2.4 | 370.5 31.5 20 3.4 | 393.0 33.5 20 3.0 | 413.3 33.9 20 4.5 | 427.1 37.4 20 4.1 | 438.9 39.0 20 4.2 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 297.9 23.2 20 -0.8 | 324.7 26.1 20 -2.9 | 348.3 30.2 20 -2.8 | 365.3 33.5 20 -4.2 | 383.9 36.0 20 -2.9 | 399.6 35.5 20 -2.6 | 410.4 40.6 20 -2.6 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
90 | 97 | 104 | 111 | 118 | 125 | 128 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 431.7 46.1 20 - | 434.7 44.9 20 - | 441.9 43.5 20 - | 453.5 48.3 20 - | 464.3 49.9 20 - | 473.8 52.6 20 - | 484.1 58.9 8 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 440.9 44.6 20 2.1 | 444.3 45.0 20 2.2 | 452.5 46.6 20 2.4 | 463.5 46.1 20 2.2 | 477.7 47.1 20 2.9 | 492.3 48.2 20 3.9 | 494.2 58.5 11 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 448.1 41.2 20 3.8 | 452.0 38.8 20 4.0 | 456.2 39.7 20 3.2 | 471.1 40.7 20 3.9 | 483.8 43.3 20 4.2 | 494.5 45.3 20 4.4 | 515.1 42.9 9 6.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 415.5 40.8 20 -3.8 | 424.9 41.7 20 -2.3 | 428.4 41.3 20 -3.1 | 442.0 42.1 20 -2.5 | 451.7 42.8 20 -2.7 | 460.8 41.2 20 -2.8 | 463.5 45.3 12 -4.3 |
Sex: Male |
|
| Day(s) Relative to Start Date |
|
129 [G] | 130 [G] | 131 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 465.0 62.2 6 - | 475.3 45.0 9 - | - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 504.9 36.5 7 8.6 | 502.0 45.0 7 5.6 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 514.8 41.9 5 10.7 | 471.0 41.8 8 -0.9 | 485.5n 36.8 4 - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 476.3 43.3 8 2.4 | 436.3 10.1 3 -8.2 | 529.0n - 1 - |
Table 8 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Prior to Mating
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 5.1 20 - | - - - - | 66.2 7.1 20 - | 103.6 9.0 20 - | 136.1 9.5 20 - | 156.9 10.6 20 - | 174.8 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 53.8 7.9 19 2.4 | 57.3n 4.0 3 - | 66.6 10.4 18 0.7 | 103.4 13.9 20 -0.1 | 133.7 13.8 20 -1.7 | 155.3 14.0 20 -1.0 | 173.5 14.1 20 -0.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 4.4 20 6.0 | 68.0n - 1 - | 71.6 7.5 19 8.3 | 109.3 8.1 20 5.5 | 143.2 9.6 20 5.3 | 164.1 10.0 20 4.6 | 180.9 11.8 20 3.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 51.3 4.3 15 -2.3 | 56.3n 6.6 7 - | 64.6 7.4 18 -2.4 | 102.8 10.3 20 -0.7 | 136.6 9.8 20 0.4 | 157.6 13.1 20 0.4 | 176.8 13.7 20 1.1 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.6 11.3 20 - | 204.2 11.0 20 - | 210.9 11.8 20 - | 218.6 12.6 20 - | 227.6 11.8 20 - | 232.5 11.5 20 - | 236.2 12.5 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 188.1 15.8 20 -0.8 | 200.3 15.1 20 -1.9 | 209.8 15.4 20 -0.5 | 218.7 14.7 20 0.1 | 226.3 16.5 20 -0.6 | 230.7 18.2 20 -0.8 | 232.7 17.3 20 -1.5 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 199.2 12.4 20 5.1 | 211.3 13.2 20 3.5 | 220.1 14.6 20 4.4 | 227.0 15.2 20 3.9 | 236.9 15.5 20 4.1 | 243.1 16.4 20 4.5 | 246.7 17.5 20 4.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 195.6 14.1 20 3.2 | 203.7 14.7 20 -0.2 | 212.2 15.4 20 0.6 | 219.5 16.9 20 0.4 | 231.0 16.5 20 1.5 | 236.4 18.1 20 1.7 | 237.4 17.1 20 0.5 |
Sex: Female |
| Day(s) Relative to Start Date |
90 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 240.0 13.4 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 239.6 16.7 20 -0.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 251.2 17.1 20 4.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 242.7 18.8 20 1.1 |
Table 9 - Summary of Body Weights (g)
F1 Animals - Cohort 1B - Gestation
Sex: Female |
|
| Day(s) Relative to Mating (Litter: A) |
| |
0 | 7 | 14 | 20 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 241.9 12.7 19 - | 261.8 15.3 19 - | 284.1 15.0 19 - | 337.5 18.9 19 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 238.8 14.3 16 -1.3 | 258.2 14.3 16 -1.4 | 277.9 14.9 16 -2.2 | 334.6 19.9 16 -0.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 253.5 16.6 19 4.8 | 271.6 16.9 19 3.8 | 291.6 16.6 19 2.6 | 348.9 19.3 19 3.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 241.1 16.7 20 -0.3 | 260.8 20.7 20 -0.4 | 278.0 23.3 20 -2.1 | 336.5 32.1 20 -0.3 |
Table 10 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | |
| Sex | 7 | |||||
| 1F | Mean | 264.2 | 268.9 | 276.9 | 290.6 | 284.2 |
| SD | 14.4 | 12.9 | 14.9 | 16.8 | 17.9 | |
| N | 18 | 17 | 18 | 18 | 18 | |
| Mean | 262.3 | 271.3 | 275.8 | 288.6 | 286.3 | |
| 2F | SD | 14.5 | 15 | 14.2 | 16.6 | 15.4 |
| N | 16 | 15 | 16 | 16 | 16 | |
| %Diff G1 | -0.7 | 0.9 | -0.4 | -0.7 | 0.8 | |
| Mean | 278.4a | 288.5b | 292.6a | 306.2a | 302.1a | |
| SD | 18.9 | 16.6 | 15.7 | 15.6 | 16.2 | |
| 3F | N | 19 | 19 | 19 | 19 | 19 |
| %Diff G1 | 5.4 | 7.3 | 5.6 | 5.4 | 6.3 | |
| Mean | 265.9 | 276.3 | 283.6 | 292.4 | 291.5 | |
| 4F | SD | 21.6 | 24.5 | 20.3 | 21.7 | 22.9 |
| N | 20 | 19 | 20 | 20 | 19 | |
| %Diff G1 | 0.6 | 2.7 | 2.4 | 0.6 | 2.6 |
Table 11 - Summary of Body Weights (g)
F1 Animals - Cohort 2A
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 4.7 10 - | - - - - | 68.4 8.0 10 - | 115.1 11.9 10 - | 169.5 12.1 10 - | 219.4 13.6 10 - | 269.5 18.2 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.6 5.5 9 -0.1 | 59.0n 7.1 2 - | 67.0 8.9 9 -2.0 | 111.0 12.0 10 -3.6 | 160.7 12.0 10 -5.2 | 205.3 14.3 10 -6.4 | 250.8 18.2 10 -6.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 58.5 5.9 10 7.1 | 76.0n - 1 - | 74.7 10.9 9 9.2 | 122.9 14.3 10 6.8 | 177.1 16.3 10 4.5 | 224.4 19.6 10 2.3 | 273.9 23.0 10 1.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 55.4 3.6 9 1.5 | 59.0n 7.1 2 - | 67.3 9.2 9 -1.6 | 112.4 11.1 10 -2.3 | 164.3 12.4 10 -3.1 | 211.4 15.1 10 -3.6 | 259.5 21.4 10 -3.7 |
Sex: Male |
|
|
| Day(s) Relative to Start Date |
|
| |
41 | 48 | 55 | 58 | 59 | 60 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 308.0 21.9 10 - | 342.0 24.0 10 - | 368.4 25.7 10 - | 369.8 24.4 4 - | 396.8 32.0 5 - | 405.0 37.8 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 287.9 19.6 10 -6.5 | 320.4 24.2 10 -6.3 | 347.9 26.8 10 -5.6 | 376.8 42.4 4 1.9 | 369.8 20.2 6 -6.8 | 360.5 16.2 4 -11.0 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 313.5 26.9 10 1.8 | 348.1 27.9 10 1.8 | 372.8 25.9 10 1.2 | 403.3 32.3 6 9.1 | 404.0 54.1 3 1.8 | 369.5 14.5 4 -8.8 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 298.6 22.8 10 -3.1 | 326.5 27.2 10 -4.5 | 347.3 31.0 10 -5.7 | 362.0 46.0 4 -2.1 | 376.9 25.2 7 -5.0 | 341.5n 0.7 2 -15.7 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 51.5 4.2 10 - | - - - - | 62.9 6.5 10 - | 98.9 9.8 10 - | 128.6 9.3 10 - | 149.3 8.6 10 - | 166.7 9.1 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.2 2.9 9 5.3 | 54.0n 2.8 2 - | 65.4 5.5 9 4.0 | 100.5 8.6 10 1.6 | 131.3 8.2 10 2.1 | 155.1 8.6 10 3.9 | 172.1 10.8 10 3.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 7.4 10 8.2 | 76.0n - 1 - | 68.8 10.0 9 9.3 | 108.9 13.2 10 10.1 | 145.1** 12.9 10 12.8 | 167.6** 15.5 10 12.3 | 186.0** 18.3 10 11.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 53.6 3.7 9 4.0 | 57.0n 5.7 2 - | 63.2 5.3 9 0.5 | 100.5 6.2 10 1.6 | 135.7 10.3 10 5.5 | 159.2 10.4 10 6.6 | 179.3 12.7 10 7.6 |
Sex: Female |
|
| Day(s) Relative to Start Date |
|
| ||
41 [G] | 48 [G] | 55 [G] | 58 [G] | 59 [G1] | 60 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 179.8 10.1 10 - | 193.9 9.9 10 - | 203.7 10.7 10 - | 204.8 13.1 4 - | 214.0 12.7 5 - | 214.3 12.2 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 185.7 12.3 10 3.3 | 199.7 13.1 10 3.0 | 210.3 14.1 10 3.2 | 206.8 12.0 4 1.0 | 218.0 20.1 6 1.9 | 210.8 12.0 4 -1.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 202.3** 20.4 10 12.5 | 215.0** 20.3 10 10.9 | 223.8* 23.4 10 9.9 | 237.2 33.5 6 15.8 | 247.0 47.1 3 15.4 | 223.5 14.2 4 4.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 198.6* 14.7 10 10.5 | 210.7* 14.0 10 8.7 | 218.6 13.4 10 7.3 | 223.0 22.0 4 8.9 | 229.9 10.5 7 7.4 | 220.7 6.0 3 3.0 |
Tables 12. Summary of Clinical Chemistry Values
F0 Animals - Males. Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G1] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 71.4 12.3 10 - | 51.0 11.7 10 - | 128.5 12.4 10 - | 1.5 0.0 10 - | 217.1 68.4 10 - | 1.25 0.00 10 - | 7.35 0.66 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 74.6 10.3 9 1.04 | 49.1 12.0 9 0.96 | 126.0 25.8 9 0.98 | 1.5 0.0 9 1.00 | 292.7 163.0 9 1.35 | 1.25 0.00 9 1.00 | 6.90 0.68 9 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 60.5 * 4.9 10 0.85 | 44.5 7.8 10 0.87 | 125.1 32.5 10 0.97 | 1.5 0.0 10 1.00 | 213.2 70.8 10 0.98 | 1.25 0.00 10 1.00 | 7.16 0.35 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.6 * 5.4 10 0.86 | 36.8 14.1 9 0.72 | 115.4 28.5 10 0.90 | 1.5 0.0 10 1.00 | 196.8 60.0 10 0.91 | 1.25 0.00 10 1.00 | 6.94 0.68 10 0.94 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.2 3.2 10 - | 8.251 0.699 10 - | 1.789 0.249 10 - | 1.949 0.657 10 - | 66.29 1.89 10 - | 40.36 1.53 10 - | 25.93 2.18 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 32.1 3.5 9 0.94 | 8.778 1.195 9 1.06 | 1.853 0.393 9 1.04 | 1.933 0.681 9 0.99 | 66.09 3.31 9 1.00 | 41.22 2.83 9 1.02 | 24.87 2.61 9 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 * 2.9 10 0.87 | 8.113 0.521 10 0.98 | 2.170 * 0.404 10 1.21 | 1.594 0.199 10 0.82 | 66.67 2.46 10 1.01 | 41.93 1.91 10 1.04 | 24.74 2.11 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 31.5 3.6 10 0.92 | 8.208 0.634 10 0.99 | 2.327 ** 0.270 10 1.30 | 1.457 0.567 10 0.75 | 68.21 2.50 10 1.03 | 43.17 * 2.11 10 1.07 | 25.04 2.86 10 0.97 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.58 0.18 10 - | 2.618 0.059 10 - | 141.1 1.3 10 - | 4.98 0.29 10 - | 103.7 1.2 10 - | 1.404 0.193 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.69 0.22 9 1.07 | 2.637 0.076 9 1.01 | 141.4 1.9 9 1.00 | 4.90 0.34 9 0.98 | 103.7 1.4 9 1.00 | 1.531 0.178 9 1.09 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.71 0.17 10 1.08 | 2.654 0.055 10 1.01 | 140.3 2.2 10 0.99 | 4.97 0.15 10 1.00 | 102.3 1.8 10 0.99 | 1.545 0.152 10 1.10 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.75 0.28 10 1.11 | 2.716 ** 0.069 10 1.04 | 141.3 1.7 10 1.00 | 5.02 0.29 10 1.01 | 101.4 ** 2.0 10 0.98 | 1.591 0.187 10 1.13 |
Tables 13. Summary of Clinical Chemistry Values
F0 Animals - Females. Day: 115 Relative to Start Date
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 131.2 28.7 9 - | 117.1 17.5 9 - | 223.4 49.2 9 - | 1.5 0.0 9 - | 630.0 384.5 9 - | 1.25 0.00 9 - | 10.50 1.01 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 105.3 10.9 8 0.80 | 110.0 18.4 8 0.94 | 200.3 73.8 7 0.90 | 1.5 0.0 8 1.00 | 356.8 132.6 8 0.57 | 1.25 0.00 8 1.00 | 10.11 0.94 8 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 116.1 21.8 9 0.88 | 105.7 14.1 9 0.90 | 157.1 38.5 9 0.70 | 1.5 0.0 9 1.00 | 614.7 356.8 9 0.98 | 1.25 0.00 9 1.00 | 9.38 0.84 9 0.89 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 106.4 25.3 10 0.81 | 100.4 21.0 10 0.86 | 173.2 49.5 10 0.78 | 1.5 0.0 9 1.00 | 428.3 244.8 9 0.68 | 1.25 0.00 9 1.00 | 10.57 1.26 9 1.01 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G1] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 29.4 3.2 9 - | 4.740 1.150 9 - | 2.828 0.449 9 - | 0.589 0.061 9 - | 56.30 2.58 9 - | 40.21 1.79 9 - | 16.09 2.22 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 24.6 * 4.2 8 0.84 | 5.008 0.434 8 1.06 | 2.641 0.493 8 0.93 | 0.461 0.140 8 0.78 | 55.21 2.41 8 0.98 | 39.51 2.62 8 0.98 | 15.70 0.72 8 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 24.0 * 2.5 8 0.82 | 4.863 0.808 8 1.03 | 2.889 0.655 9 1.02 | 0.535 0.177 8 0.91 | 56.38 2.85 8 1.00 | 40.59 2.45 9 1.01 | 16.19 1.63 8 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 25.6 4.6 9 0.87 | 5.003 0.815 9 1.06 | 2.827 0.925 9 1.00 | 0.479 0.115 9 0.81 | 58.48 3.36 9 1.04 | 42.81 2.93 10 1.06 | 15.68 1.98 9 0.97 |
Sex: Female |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.53 0.34 9 - | 2.680 0.100 9 - | 140.7 2.1 9 - | 5.12 0.42 9 - | 99.8 2.2 9 - | 1.708 0.538 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.54 0.23 8 1.00 | 2.621 0.139 8 0.98 | 140.0 3.5 8 1.00 | 5.05 0.57 8 0.99 | 100.1 1.9 8 1.00 | 1.649 0.518 8 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.50 0.29 8 0.99 | 2.694 0.096 9 1.01 | 140.1 1.3 9 1.00 | 5.24 0.62 9 1.02 | 99.3 1.7 9 1.00 | 1.538 0.510 8 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.78 0.47 9 1.10 | 2.781 0.183 9 1.04 | 140.1 2.3 10 1.00 | 5.19 0.42 10 1.01 | 97.8 2.9 10 0.98 | 1.949 0.766 9 1.14 |
Tables 14. Summary of Clinical Chemistry Values
F1 Animals - Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 68.5 6.0 10 - | 57.6 11.4 10 - | 153.4 23.5 10 - | 1.5 0.0 9 - | 239.9 42.0 9 - | 1.25 0.00 9 - | 7.02 0.68 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 63.0 7.2 10 0.92 | 52.0 7.9 10 0.90 | 153.5 23.8 10 1.00 | 1.5 0.0 10 1.00 | 239.2 101.7 10 1.00 | 1.25 0.00 10 1.00 | 6.32 0.62 10 0.90 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 61.3 * 5.3 10 0.89 | 49.9 8.6 10 0.87 | 143.2 27.9 10 0.93 | 1.5 0.0 10 1.00 | 210.5 55.4 10 0.88 | 1.25 0.00 10 1.00 | 6.84 0.59 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.2 * 4.0 10 0.89 | 42.5 ** 13.0 10 0.74 | 141.5 23.9 10 0.92 | 1.5 0.0 10 1.00 | 174.9 37.4 10 0.73 | 1.25 0.00 10 1.00 | 7.24 0.85 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 30.8 4.0 9 - | 9.268 0.551 9 - | 1.723 0.237 9 - | 2.458 1.001 9 - | 66.48 1.47 9 - | 41.30 1.39 10 - | 25.16 1.24 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 28.1 3.8 10 0.91 | 8.529 ** 0.254 10 0.92 | 1.894 0.316 10 1.10 | 2.030 0.674 10 0.83 | 66.44 1.28 10 1.00 | 41.71 2.18 10 1.01 | 24.73 2.00 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 2.6 10 0.97 | 8.735 0.590 10 0.94 | 2.007 0.325 10 1.16 | 1.784 0.635 10 0.73 | 65.87 2.39 10 0.99 | 41.11 2.06 10 1.00 | 24.76 2.03 10 0.98 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.6 3.7 10 1.06 | 8.617 * 0.503 10 0.93 | 2.147 * 0.325 10 1.25 | 1.522 0.615 10 0.62 | 65.97 1.76 10 0.99 | 42.49 1.31 10 1.03 | 23.48 1.78 10 0.93 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G1] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.64 0.10 9 - | 2.592 0.033 9 - | 141.0 0.8 10 - | 5.09 0.30 10 - | 100.5 0.5 10 - | 1.747 0.191 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.70 0.19 10 1.03 | 2.620 0.064 10 1.01 | 142.0 1.2 10 1.01 | 5.12 0.30 10 1.01 | 101.7 * 0.8 10 1.01 | 2.034 ** 0.122 10 1.16 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.67 0.19 10 1.02 | 2.626 0.074 10 1.01 | 141.1 1.5 10 1.00 | 4.99 0.20 10 0.98 | 101.3 1.2 10 1.01 | 1.968 ** 0.179 10 1.13 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.83 0.17 10 1.11 | 2.657 0.064 10 1.02 | 141.6 2.0 10 1.00 | 4.95 0.18 10 0.97 | 99.9 1.6 10 0.99 | 2.065 ** 0.116 10 1.18 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 78.0 10.7 7 - | 55.9 14.4 7 - | 100.6 26.1 7 - | 1.5 0.0 7 - | 295.7 114.0 7 - | 1.25 0.00 7 - | 8.09 1.15 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 71.9 7.5 9 0.92 | 54.6 17.9 9 0.98 | 93.2 23.3 9 0.93 | 1.5 0.0 9 1.00 | 242.9 83.5 9 0.82 | 1.25 0.00 9 1.00 | 7.43 0.78 9 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 68.3 6.1 10 0.88 | 49.9 13.2 10 0.89 | 94.3 24.1 10 0.94 | 1.5 0.0 10 1.00 | 259.2 98.3 10 0.88 | 1.25 0.00 10 1.00 | 7.70 0.89 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 73.7 9.9 10 0.94 | 59.8 26.8 10 1.07 | 135.4 38.1 10 1.35 | 2.2 1.5 10 1.47 | 296.8 105.5 10 1.00 | 1.25 0.00 10 1.00 | 7.66 1.24 10 0.95 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.0 3.5 7 - | 8.577 0.866 7 - | 1.589 0.267 7 - | 1.421 0.304 7 - | 65.96 2.49 7 - | 45.56 2.60 7 - | 20.40 2.41 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 34.8 4.5 9 1.02 | 7.801 0.566 9 0.91 | 1.512 0.297 9 0.95 | 1.021 0.261 9 0.72 | 67.76 2.46 9 1.03 | 46.70 1.56 9 1.03 | 21.06 1.80 9 1.03 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 32.7 4.0 10 0.96 | 8.161 0.464 10 0.95 | 1.647 0.235 10 1.04 | 1.268 0.529 10 0.89 | 68.21 2.48 10 1.03 | 46.77 2.62 10 1.03 | 21.44 2.18 10 1.05 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.4 4.5 10 0.95 | 8.082 0.391 10 0.94 | 1.913 * 0.234 10 1.20 | 0.978 0.268 10 0.69 | 67.82 2.61 10 1.03 | 46.97 2.19 10 1.03 | 20.85 1.87 10 1.02 |
Sex: Female |
|
| Reporting Biochemistry |
|
| ||
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.27 0.35 7 - | 2.574 0.035 7 - | 140.3 1.1 7 - | 4.43 0.42 7 - | 103.6 1.4 7 - | 1.227 0.104 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.24 0.21 9 0.99 | 2.581 0.060 9 1.00 | 140.2 1.2 9 1.00 | 4.78 0.29 9 1.08 | 102.9 1.2 9 0.99 | 1.436 0.206 9 1.17 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.18 0.30 10 0.96 | 2.591 0.053 10 1.01 | 140.6 0.8 10 1.00 | 4.45 0.20 10 1.00 | 104.1 1.3 10 1.01 | 1.379 0.205 10 1.12 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.30 0.28 10 1.01 | 2.664 ** 0.039 10 1.03 | 140.4 1.1 10 1.00 | 4.86 * 0.27 10 1.10 | 102.2 0.9 10 0.99 | 1.312 0.263 10 1.07 |
Table 15. Summary of Coagulation Values
F0 Animals – Males, Day: 137 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.55 0.20 10 - | 13.51 1.35 10 - | 2.278 0.177 10 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.43 0.15 10 0.99 | 12.52 1.72 10 0.93 | 2.282 0.122 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.30 ** 0.14 10 0.98 | 12.35 1.14 10 0.91 | 2.286 0.185 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.26 ** 0.12 10 0.97 | 11.77 1.51 10 0.87 | 2.454 * 0.114 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 16. Summary of Coagulation Values
F0 Animals – Females, Day: 115 Relative to Start Date
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.19 0.21 9 - | 11.17 1.22 9 - | 1.713 0.292 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.48 0.34 10 1.03 | 12.17 1.55 10 1.09 | 1.612 0.157 10 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.27 0.32 9 1.01 | 11.82 1.49 9 1.06 | 1.743 0.316 9 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.39 0.73 10 1.02 | 11.16 2.64 10 1.00 | 1.682 0.425 10 0.98 |
[G] - Anova & Dunnett
Table 17. Summary of Coagulation Values
F1 Animals - Cohort 1A, Day: 70 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.50 0.14 9 - | 12.27 1.80 9 - | 2.580 0.172 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.40 0.11 10 0.99 | 12.79 2.10 10 1.04 | 2.516 0.141 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.34 * 0.14 10 0.98 | 12.09 2.30 10 0.99 | 2.655 0.219 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.33 * 0.05 9 0.98 | 11.51 2.25 9 0.94 | 2.707 0.250 9 1.05 |
[G] - Anova & Dunnett: * = p ≤ 0.05
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.31 0.26 9 - | 12.21 1.52 8 - | 2.116 0.251 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.17 0.17 9 0.99 | 12.33 1.57 9 1.01 | 2.128 0.172 9 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.15 0.14 10 0.98 | 11.65 0.99 10 0.95 | 2.186 0.194 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.02 ** 0.17 10 0.97 | 11.71 1.65 10 0.96 | 2.182 0.099 10 1.03 |
[G] - Anova & Dunnett: ** = p ≤ 0.01
Table 18. Summary of Hematology Values
F0 Animals – Males Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G1] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 4.651 0.478 10 - | 0.961 0.257 10 - | 3.480 0.289 10 - | 0.065 0.014 10 - | 0.116 0.046 10 - | 0.008 0.004 10 - | 0.019 0.006 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 4.340 0.805 10 0.93 | 0.769 0.202 10 0.80 | 3.390 0.784 10 0.97 | 0.062 0.018 10 0.95 | 0.096 0.024 10 0.83 | 0.006 0.005 10 0.75 | 0.016 0.008 10 0.84 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.469 0.783 10 0.96 | 0.687 * 0.169 10 0.71 | 3.605 0.697 10 1.04 | 0.057 0.021 10 0.88 | 0.089 0.023 10 0.77 | 0.010 0.000 10 1.25 | 0.019 0.007 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.192 0.960 10 1.12 | 0.854 0.199 10 0.89 | 4.126 0.908 10 1.19 | 0.079 0.028 10 1.22 | 0.092 0.040 10 0.79 | 0.009 0.006 10 1.13 | 0.027 0.013 10 1.42 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 8.813 0.449 10 - | 153.6 4.7 10 . | 0.4464 0.0164 10 - | 50.68 1.37 10 - | 17.43 0.57 10 - | 343.9 7.0 10 - | 11.88 0.31 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.630 0.303 10 0.98 | 150.7 6.5 10 0.98 | 0.4441 0.0155 10 0.99 | 51.52 2.26 10 1.02 | 17.46 1.03 10 1.00 | 338.9 7.5 10 0.99 | 12.00 0.48 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.572 0.317 10 0.97 | 149.6 5.2 10 0.97 | 0.4388 0.0129 10 0.98 | 51.25 1.89 10 1.01 | 17.47 0.71 10 1.00 | 341.1 6.6 10 0.99 | 12.01 0.52 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.573 0.220 10 0.97 | 147.4 5.4 10 0.96 | 0.4364 0.0139 10 0.98 | 50.92 1.39 10 1.00 | 17.17 0.55 10 0.99 | 337.5 4.5 10 0.98 | 12.16 0.48 10 1.02 |
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 781.6 146.2 10 - | 155.83 19.62 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 765.3 76.5 10 0.98 | 142.51 41.33 10 0.91 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 760.4 92.6 10 0.97 | 160.37 18.17 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 883.4 160.3 10 1.13 | 171.47 21.32 10 1.10 |
Table 19. Summary of Hematology Values
F0 Animals – Females Day: 115 Relative to Start Date
Sex: Female |
| Reporting Hematology | ||||||
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.168 1.072 8 - | 1.669 0.559 8 - | 3.265 0.790 8 - | 0.109 0.031 8 - | 0.095 0.038 8 - | 0.011 0.006 8 - | 0.023 0.009 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.529 0.603 10 1.07 | 1.737 0.408 10 1.04 | 3.564 0.620 10 1.09 | 0.113 0.025 10 1.04 | 0.077 0.039 10 0.81 | 0.012 0.008 10 1.07 | 0.025 0.010 10 1.11 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 5.866 1.252 10 1.14 | 1.903 0.424 10 1.14 | 3.694 0.894 10 1.13 | 0.143 0.060 10 1.31 | 0.091 0.038 10 0.96 | 0.013 0.007 10 1.16 | 0.026 0.013 10 1.16 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.570 0.802 10 1.08 | 1.762 0.315 10 1.06 | 3.555 0.672 10 1.09 | 0.144 0.051 10 1.32 | 0.073 0.013 10 0.77 | 0.008 0.004 10 0.71 | 0.028 0.011 10 1.24 |
Sex: Female |
| Reporting Hematology | ||||||
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.958 0.449 8 - | 157.3 3.4 8 . | 0.4679 0.0124 8 - | 58.88 2.37 8 - | 19.80 0.92 8 - | 336.4 5.2 8 - | 12.00 1.06 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.678 0.380 10 0.96 | 153.3 5.2 10 0.97 | 0.4515 0.0208 10 0.97 | 58.86 1.56 10 1.00 | 19.99 0.77 10 1.01 | 339.8 8.3 10 1.01 | 11.84 0.79 10 0.99 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.466 0.606 9 0.94 | 152.7 5.1 10 0.97 | 0.4468 0.0194 9 0.95 | 59.99 2.69 9 1.02 | 20.48 1.21 9 1.03 | 341.6 7.1 9 1.02 | 11.82 0.97 10 0.99 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.528 0.566 10 0.95 | 151.3 8.8 10 0.96 | 0.4428 0.0316 10 0.95 | 58.85 1.83 10 1.00 | 20.10 1.07 10 1.02 | 341.5 10.8 10 1.02 | 12.17 1.05 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 894.6 119.2 8 - | 177.99 20.44 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 794.3 132.8 10 0.89 | 178.18 31.61 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 872.4 121.5 9 0.98 | 161.09 22.77 10 0.91 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 833.4 177.2 10 0.93 | 173.25 30.61 10 0.97 |
Table 20. Summary of Hematology Values
F1 Animals – Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 6.989 1.038 10 - | 0.930 0.183 10 - | 5.762 0.940 10 - | 0.121 0.043 10 - | 0.108 0.036 10 - | 0.008 0.009 10 - | 0.059 0.018 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.203 0.765 10 1.03 | 0.970 0.322 10 1.04 | 5.976 0.743 10 1.04 | 0.099 0.031 10 0.82 | 0.091 0.031 10 0.84 | 0.011 0.006 10 1.38 | 0.054 0.023 10 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.256 0.780 10 1.04 | 0.993 0.193 10 1.07 | 6.050 0.666 10 1.05 | 0.086 0.030 10 0.71 | 0.074 0.023 10 0.69 | 0.007 0.005 10 0.88 | 0.047 0.009 10 0.80 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.090 * 0.978 10 1.16 | 1.081 0.253 10 1.16 | 6.736 1.071 10 1.17 | 0.115 0.030 10 0.95 | 0.080 0.032 10 0.74 | 0.011 0.006 10 1.38 | 0.064 0.023 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Hematology |
|
|
| |
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 9.078 0.461 10 - | 163.2 4.9 10 . | 0.4747 0.0207 10 - | 52.33 1.52 10 - | 17.99 0.90 10 - | 343.7 10.9 10 - | 11.47 0.49 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.551 * 0.488 10 0.94 | 156.9 * 5.4 10 0.96 | 0.4471 ** 0.0181 10 0.94 | 52.33 2.09 10 1.00 | 18.37 0.87 10 1.02 | 350.8 6.2 10 1.02 | 11.59 0.52 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.522 ** 0.298 10 0.94 | 157.1 * 4.1 10 0.96 | 0.4519 * 0.0171 10 0.95 | 53.06 1.54 10 1.01 | 18.44 0.54 10 1.03 | 347.7 7.0 10 1.01 | 11.63 0.51 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.340 ** 0.285 10 0.92 | 154.7 ** 4.4 10 0.95 | 0.4456 ** 0.0166 10 0.94 | 53.45 1.51 10 1.02 | 18.56 0.67 10 1.03 | 347.4 7.6 10 1.01 | 11.84 0.32 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 763.2 131.0 10 - | 193.85 15.23 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 876.8 89.5 10 1.15 | 171.56 23.85 10 0.89 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 899.7 76.1 10 1.18 | 198.49 23.89 10 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 823.3 144.5 10 1.08 | 208.78 30.71 10 1.08 |
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G1] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G1] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.104 1.013 9 - | 0.587 0.094 9 - | 4.321 0.997 9 - | 0.070 0.018 9 - | 0.081 0.019 9 - | 0.003 0.005 9 - | 0.038 0.010 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.207 1.111 9 1.02 | 0.671 0.142 9 1.14 | 4.371 1.058 9 1.01 | 0.053 0.017 9 0.76 | 0.074 0.028 9 0.92 | 0.004 0.005 9 1.33 | 0.029 0.014 9 0.76 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.656 0.870 10 0.91 | 0.708 0.238 10 1.21 | 3.780 0.791 10 0.87 | 0.060 0.021 10 0.86 | 0.067 0.020 10 0.83 | 0.003 0.005 10 0.90 | 0.034 0.008 10 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.515 1.387 10 1.08 | 0.681 0.323 10 1.16 | 4.640 1.417 10 1.07 | 0.076 0.039 10 1.09 | 0.079 0.031 10 0.97 | 0.004 0.005 10 1.20 | 0.037 0.014 10 0.98 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.842 0.436 9 - | 151.6 5.3 9 . | 0.4248 0.0164 9 - | 54.23 1.82 9 - | 19.38 0.95 9 - | 356.8 8.1 9 - | 10.70 0.38 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.868 0.225 9 1.00 | 148.8 6.5 9 0.98 | 0.4244 0.0173 9 1.00 | 53.91 1.31 9 0.99 | 18.89 0.64 9 0.97 | 350.7 8.5 9 0.98 | 10.91 0.36 9 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.867 0.362 10 1.00 | 149.7 5.4 10 0.99 | 0.4244 0.0143 10 1.00 | 54.01 2.25 10 1.00 | 19.06 0.93 10 0.98 | 352.6 4.3 10 0.99 | 10.68 0.35 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.744 0.368 10 0.99 | 144.3 5.5 10 0.95 | 0.4116 0.0156 10 0.97 | 53.20 1.94 10 0.98 | 18.64 0.75 10 0.96 | 350.5 6.8 10 0.98 | 10.82 0.33 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 833.0 141.8 9 - | 183.07 34.30 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 801.3 145.2 8 0.96 | 178.02 22.41 9 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 796.3 97.2 9 0.96 | 185.18 28.61 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 910.6 58.3 10 1.09 | 177.80 38.40 10 0.97 |
Table 21. Summary of Reproductive Performance
F0 Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 25 25 25 5.3 5.1 25 - 19 76.0 6 24.0 100.0 25/25 | 25 25 25 3.2 2.5 25 -39.4 24 96.0 1 4.0 100.0 25/25 | 25 25 25 2.7 2.6 25 -49.2 24 96.0 1 4.0 100.0 25/25 | 25 24 24 3.0 2.7 24 -43.2 23 95.8 1 4.2 100.0 24/24 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 22. Summary of Reproductive Performance
F1 Animals - Cohort 1B Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 20 20 20 2.7 1.0 20 - 20 100.0 0 0.0 100.0 20/20 | 20 20 20 3.9 3.7 20 45.3 18 90.0 2 10.0 100.0 20/20 | 20 20 20 3.5 2.7 20 32.1 19 95.0 1 5.0 100.0 20/20 | 20 20 20 2.5 1.2 20 -7.5 20 100.0 0 0.0 100.0 20/20 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 23. Mating Performance and Fertility
F0 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 6(1)* | 4 | 8 | 7(1) |
2 | 2 | 5 | 6 | 5 |
3 | 6 | 8 | 7 | 3 |
4 | 5 | 7 | 3 | 8 |
No clear indication of mating | 6(1) | 1 | 1 | 1 |
Mean number of nights to positive mating sign | 2.5 | 2.8 | 2.2 | 2.5 |
Number passing one estrus | 0 | 0 | 0 | 0 |
Number of males paired | 25 | 25 | 25 | 24 |
Number of siring males | 24 | 25 | 25 | 23 |
Male Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Male Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Male Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
Female Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Female Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Female Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
*Includes Animal 1524 that had 2 implant sites (no live or dead pups)
Table 24. Mating Performance and Fertility
F1 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 3 | 5 | 2 | 7 |
2 | 5 | 2 | 4 | 2 |
3 | 9 | 5(2) | 8 | 6 |
4 | 2 | 5 | 4 | 5 |
5 | 1(1) | 1 | 0 | 0 |
6 | 0 | 0 | 1 | 0 |
No clear indication of mating | 0 | 2(2) | 1(1) | 0 |
Mean number of nights to positive mating sign | 2.7 | 2.7 | 2.9 | 2.5 |
Number passing one estrus | 0 | 1 | 1 | 0 |
Number of males paired | 20 | 20 | 20 | 20 |
Number of siring males | 19 | 16 | 19 | 20 |
Male Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Male Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Male Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Female Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Female Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Female Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Table 25. Summary of Duration of Gestation and Overall Litter Performance
F0 Animals
F0 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 24 | 25 | 25 | 23 |
Duration of Gestation (Days) 21 |
1 |
1 |
0 |
0 |
22 | 17 | 17 | 23 | 20 |
23 | 0 | 6 | 1 | 2 |
No clear indication of mating | 6 | 1 | 1 | 1 |
Mean Duration | 21.9 | 22.2 | 22.0 | 22.1 |
Number of females producing a live litter | 23 | 25 | 25 | 23 |
Gestation index as % | 96 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.0 ± 3.4 | 12.5 ± 2.4 | 12.6 ± 1.9 | 12.9 ± 1.6 |
Mean total number of pups born per litter ± standard deviation | 11.0 ± 3.0 | 11.2 ± 2.3 | 11.0 ± 2.5 | 11.0 ± 3.3 |
Mean number of live pups per litter ± standard deviation: Lactation Day 1 |
11.0 ± 3.0 |
11.1 ± 2.2 |
10.9 ± 2.6 |
11.1 ± 2.6 |
Lactation Day 4 | 10.9 ± 3.1 | 11.2 ± 2.2 | 10.9 ± 2.6 | 11.0 ± 2.7 |
Lactation Day 4a | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 7 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8± 0.7 | 7.7 ± 0.9 |
Lactation Day 14 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 21 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Number of males on Lactation Day 1 (%) | 120 (47) | 135 (49) | 145 (53) | 116 (47) |
Number of females on Lactation Day 1 (%) | 134 (53) | 143 (51) | 127 (47) | 129 (53) |
a After cull of pups on PND 4 to standardise the size of the litter
Table 26. Summary of Duration of Gestation and Overall Litter Performance
F1 Animals
F1 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 19 | 16 | 19 | 20 |
Duration of Gestation (Days) 21 |
1 |
3 |
1 |
1 |
22 | 15 | 9 | 17 | 18 |
23 | 3 | 4 | 1 | 1 |
Mean Duration | 22.1 | 22.1 | 22.0 | 22.0 |
Number of females producing a live litter | 19 | 16 | 19 | 20 |
Gestation index as % | 100 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.6 ± 1.7 | 12.3 ± 2.3 | 13.0 ± 1.6 | 12.5 ± 2.3 |
Mean total number of pups born per litter ± standard deviation* | 11.6 ± 1.9 | 11.1 ± 2.8 | 10.5 ± 2.8 | 11.3 ± 2.2 |
Mean number of live pups per litter ± standard deviation*: Lactation Day 1 |
11.3 ± 2.2 |
11.1 ± 3.0 |
10.4 ± 2.7 |
11.1 ± 2.1 |
Lactation Day 4 | 11.3 ± 2.2 | 11.0 ± 2.9 | 10.3 ± 2.8 | 11.0 ± 2.1 |
Lactation Day 4a | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 7 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 14 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 21 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.8 ± 0.5 |
Number of males on Lactation Day 1 (%) | 96 (47) | 96 (54) | 93 (49) | 100 (45) |
Number of females on Lactation Day 1 (%) | 108 (53) | 81 (46) | 96 (51) | 121 (55) |
a After cull of pups on PND 4 to standardise the size of the litter
* Excludes Animal 1628 that had a total litter loss
Table 27. Summary of Urinalysis Values
F0 Animals - Males
Day: 131 Relative to Start Date
| Sex: Male | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G1] | |||
| 0 mg/kg/day | Mean | 4.32 | 1.0152 | 7.5 |
| Group 1 | SD | 2.94 | 0.0089 | 0.94 |
| N | 10 | 10 - | 10 - | |
| - | ||||
| 100 mg/kg/day | Mean | 5.64 | 1.0135 | 8 |
| Group 2 | SD | 2.99 | 0.0052 | 0.62 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.31 | 1 | 1.07 | |
| 300 mg/kg/day | Mean | 4.22 | 1.0215 | 8.15 |
| Group 3 | SD | 2.11 | 0.0086 | 0.63 |
| N | 10 | 10 | 10 | |
| tCtrl | 0.98 | 1.01 | 1.09 | |
| 900 mg/kg/day | Mean | 5.25 | 1.0300 ** | 7.35 0.71 |
| Group 4 | SD | 2.39 | 0.0133 | 10 |
| N | 10 | 10 | 0.98 | |
| tCtrl | 1.22 | 1.01 |
Table 28. Summary of Urinalysis Values
F0 Animals - Females
Day: 108 Relative to Start Date
| Sex: Female | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G1] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 4.85 | 1.0188 | 8.6 |
| Group 1 | SD | 2.33 | 0.0052 | 0.32 |
| N | 10 - | 10 - | 10 - | |
| 100 mg/kg/day | Mean | 5.23 | 1.0192 | 8.45 |
| Group 2 | SD | 2.96 | 0.0069 | 0.16 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.08 | 1 | 0.98 | |
| 300 mg/kg/day | Mean | 6.01 | 1.0199 | 8.2 |
| Group 3 | SD | 1.68 | 0.0039 | 0.42 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.24 | 1 | 0.95 | |
| 900 mg/kg/day | Mean | 7.05 2.58 | 1.0218 0.0051 | 8.05 * |
| Group 4 | SD | 10 | 10 | 0.72 |
| N | 1.45 | 1 | 10 | |
| tCtrl | 0.94 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05
Table 29. Summary of Urinalysis Values
F1 Animals - Cohort 1A
Day: 60 Relative to Start Date
| Sex: Male | Reporting urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 2.67 | 1.0182 | 8.11 |
| Group 1 | SD | 1.87 | 0.0078 | 0.6 |
| N | 10 - | 10 - | 9 - | |
| 100 mg/kg/day | Mean | 3.07 | 1.0168 | 7.9 |
| Group 2 | SD | 0.95 | 0.0123 | 0.61 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.15 | 1 | 0.97 | |
| 300 mg/kg/day | Mean | 1.96 | 1.0302 * | 7.11 ** 0.70 |
| Group 3 | SD | 1.25 | 0.0118 | 9 |
| N | 10 | 10 | 0.88 | |
| tCtrl | 0.73 | 1.01 | ||
| 900 mg/kg/day | Mean | 2.94 1.43 | 1.0346 ** | 6.25 ** |
| Group 4 | SD | 10 | 0.0091 | 0.42 |
| N | 1.1 | 10 | 10 | |
| tCtrl | 1.02 | 0.77 |
Day: 60 Relative to Start Date
Sex: Female |
|
| Reporting Urinalysi | s |
VOLUME (mL) [G] | SPECIFIC GRAVITY [G] | URINE pH [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.88 1.25 10 - | 1.0226 0.0117 10 - | 7.00 0.94 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 3.20 2.88 10 1.70 | 1.0161 0.0086 10 0.99 | 7.15 0.91 10 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.33 1.08 10 0.71 | 1.0333 0.0118 10 1.01 | 6.61 0.55 9 0.94 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.61 1.55 9 1.39 | 1.0267 0.0087 9 1.00 | 6.44 0.68 9 0.92 |
[G] - Anova & Dunnett
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 900 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
- Quality of whole database:
- The reproductive toxicity of the registered substance was determined in an Extended One Generation Reproductive Toxicity study using read-across to a analogue substance. The study was conducted according to the OECD 443 guideline and with GLP certification. A full study report is available. The quality of the whole database is therefore considered to be high.
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Effects on developmental toxicity
Description of key information
The test substance was tested for its prenatal developmental toxicity in two species, the rat and the rabbit both to full OECD guideline 414 and performed according to GLP
1.The test substance was tested firstly for its prenatal developmental toxicity in Wistar rats (OECD guideline 414, GLP).
The test substance was administered as an aqueous suspension to groups of 25 time-mated female Wistar rats by gavage at doses of 50, 150, and 500 mg/kg body weight/day (mg/kg bw/d) on gestation days (GD) 6 through 19. Under the conditions of this prenatal developmental toxicity study, the oral administration to pregnant Wistar rats from implantation to one day prior to the expected day of parturition (GD 6-19) at a dose of 500 mg/kg bw/d caused no evidence of maternal toxicity. In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 500 mg/kg bw/d.
2. The test susbstance was subsequently tested for its prenatal developmental toxicity in New Zealand Rabbits (OECD guideline 414, GLP).
The test substance was tested for its prenatal developmental toxicity in New Zealand White rabbits.The test substance was administered as an aqueous suspension to groups of 25 inseminated female New Zealand White rabbits orally by gavage in doses of 15, 50 and 150 mg/kg body weight/day (mg/kg bw/d) on gestation days (GD) 6 through 28. The vehicle control group, consisting of 25 females, was dosed with the vehicle 0.5% Carboxymethylcellulose suspension in drinking water (0.5% CMC) in parallel. A standard dose volume of 10 mL/kg body weight was used for each test group.
Under the conditions of this prenatal developmental toxicity study, the oral administration of the test material to pregnant New Zealand White rabbits from implantation to one day prior to the expected day of parturition (GD 6-28) caused evidence of maternal toxicity at the high-dose (150 mg/kg bw/d), such as adverse clinical findings. In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 50 mg/kg bw/d.
Since there was no evidence for toxicologically relevant adverse effects of the test substance on fetal morphology at any dose, the no observed adverse effect level (NOAEL) for prenatal developmental toxicity is the highest dose of 150 mg/kg bw/d. The rabbit is considered to be the more senstive species in this study and the lower of the NOAEL values has been taken as the definitive result.
3. The administration of the cource substance once daily (or twice daily 3 hours apart on some occasions) via
oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no
test item-related deaths. There were occasional observations of decreased activity, abnormal, un
coordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose le
vel-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body we
ight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/
day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. For F1 and F2 developmental para
meters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on
the pup numbers, survival or development.
Link to relevant study records
- Endpoint:
- developmental toxicity
- Remarks:
- extended one-generation reproductive toxicity - with F2 generation and developmental neurotoxicity (Cohorts 1A, 1B with extension, 2A and 2B)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Remarks:
- Information on target substance.
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
See attachment.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See attachment.
3. ANALOGUE APPROACH JUSTIFICATION
See attachment.
4. DATA MATRIX
See attachment.
ECHA decisions on testing proposals:
Target substance requirement:
Extended one-generation reproductive toxicity study (Annex X, Section 8.7.3.; test method: OECD TG 443) in rats, oral (gavage) route specified as follows:
- Ten weeks premating exposure duration for the parental (P0) generation;
- Dose level setting shall aim to induce systemic toxicity at the highest dose level;
- Cohort 1A (Reproductive toxicity);
- Cohort 1B (Reproductive toxicity) without extension to mate the Cohort 1B animals to produce the F2 generation.
Source substance requirement:
Additional data generated from the extension to mate the Cohort 1B to produce the F2 generation and also the Cohorts 2A and 2B (developmental neurotoxicity) was a requirement of the decision on a testing proposal for the source Hydroxycyclohexyl phenyl ketone (TPE-D-2114510002-76-01/F) and that this has been included for openness and completeness of reporting of the actual experimental study performed. - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 424. Neurotoxicity Study in Rodents
- Version / remarks:
- 21 July 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 443 Extended One-Generation Reproductive Toxicity Study
- Version / remarks:
- 25 June 2018
- Deviations:
- no
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- Han Wistar (Crl:WI(Han))
- Route of administration:
- oral: gavage
- Analytical verification of doses or concentrations:
- yes
- Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- control
- Dose / conc.:
- 100 mg/kg bw/day
- Dose / conc.:
- 300 mg/kg bw/day
- Dose / conc.:
- 900 mg/kg bw/day
- No. of animals per sex per dose:
- 10 animals per sex per dose
- Control animals:
- yes, concurrent vehicle
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- At 900 mg/kg/day, decreased activity was observed for one male on Day 2 and abnormal, unco
ordinated gait for 3 males and 4 females between Days 2 to 4. Decreased activity, abnormal, uncoord
inated gait or low carriage were occasionally noted for a further 8 females between Days 5 to 70 and
decreased activity on one occasion for each of 2 females during lactation. These findings were cons
idered to be test item-related.
Salivation and/or ploughing (also a chewing action for one female at 900 mg/kg/day) were noted for
all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, on multiple days thro
ughout the dosing period. The incidence of these findings was dose level-related.
Animal 4515F (900 mg/kg/day) had abnormal clincal observations of irregular respiratory rate, hunc
hed posture, erect fur, cold to touch, prominent backbone, abnormal gait, decreased activity and red
discharge from vagina the day after parturition (LD 1) but these mostly resolved before scheduled
termination. The clinical observations were considered to be likely due to the trauma of giving birth
rather than being test item-related.
Animal 4525F (900 mg/kg/day) had abnormal clincal observations of irregular/decreased respiratory
rate, erect fur, eyes partly closed, decreased activity up to 2-4 hours postdose on LD 7 only. No
similar findings were observed for this animal during the dosing period and as a cause could not be determined, these findings were likely related to the dosing procedure. - Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- None of the unscheduled deaths were considered to be test item-related.
On Study Day 40, approximately 3 hours after dosing, Animal 4510F given 900 mg/kg/day was found
with clinical signs of irregular respiratory rate, hunched posture, partly closed eyes, erect fur, unc
oordinated and abnormal gait and decreased activity and was euthanised. There were no findings
at necropsy or microscopically to suggest the likely cause of death. This animal had not had any
abnormal clinical signs before this (apart from salivation and ploughing) and had consistently gained
body weight.
Animals 2512F and 4502F given 100 or 900 mg/kg/day respectively, were euthanised on LD 1 or LD
0 and due to the very young age of the pups, they were also euthanised. Animal 2512F had clinical
signs of irregular respiratory rate, hunched posture, pale skin and cold to touch, erect fur, uncoor
dinated and abnormal gait in the afternoon of LD 1. Animal 4502F was found subdued, cold to touch,
prostrate/lying on side with decreased respiratory rate approximately 1 hour after dosing on LD 0. Th
ere were no findings at necropsy or microscopically to suggest the likely cause of death and both an
imals had not had any abnormal clinical signs before this (apart from salivation and ploughing for 4
502F) and had consistently gained body weight. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See Tables 1-4.
For males given 900 mg/kg/day, there was slightly lower body weight gain over the dosing period
resulting in male body weight being 3.6% lower than the concurrent control mean value by Day 134.
For females given ≥ 100 mg/kg/day, a dose level-related higher body weight gain of up to 10.3% at 90
0 mg/kg/day compared to controls was noted during the pre-pairing period and this slightly higher bo
dy weight was generally maintained throughout gestation and lactation.
There was no test item-related effect on male body weight at 100 or 300 mg/kg/day. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- There were occasional slightly higher food consumption values of up to 14.4% of controls noted for
males at 300 or 900 mg/kg/day up to Day 106. At ≥100 mg/kg/day for males from Days 106 to 134,
there was higher food consumption of up to 17.5%. For females given ≥100 mg/kg/day, there was an
overall dose level-related higher food consumption of up to 20.6% of control values during the prep
airing
period, and during gestation and lactation, food consumption was higher for females by up to
13.2% at 300 or 900 mg/kg/day only. - Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Haematology
See tables 18-19.
Coagulation
See tables 15-16.
All differences in haematology or coagulation parameters were considered not to be test item-related
based on their small magnitude, inconsistent direction, absence of a dose response, general overl
ap of individual values with the range of control values and/or were of a magnitude of variation com
monly observed in rats. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 12-13.
At ≥300 mg/kg/day for males and ≥100 mg/kg/day for females, triglycerides were lower than controls
by up to 0.75-fold, and for males only at ≥300 mg/kg/day, cholesterol higher by up to 1.30-fold. - Endocrine findings:
- effects observed, treatment-related
- Description (incidence and severity):
- There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of
necropsy in F0 adult males given 300 or 900 mg/kg/day.
There were no test item-related effects for F0 adult males given 100 mg/kg/day or F0 adult females gi
ven up to 900 mg/kg/day or for culled pups on PND 4 or unselected pups on PND 21.
There were no test item-related changes to T4 levels at up to 900 mg/kg/day for samples taken on the
morning of necropsy for the F0 generation animals, for culled pups on PND 4 or unselected pups on
PND 21.
The differences in these parameters were considered not to be test item-related based on their small
magnitude, inconsistent direction, absence of a dose response, general overlap of individual values
with the range of control and/or were of a magnitude of variation commonly observed in rats. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See tables 27-28.
All differences in urine parameters were considered not to be test item-related based on their small m
agnitude, inconsistent direction, absence of a dose response, general overlap of individual values with
the range of control and/or were of a magnitude of variation commonly observed in rats. - Immunological findings:
- no effects observed
- Description (incidence and severity):
- Splenic Immunophenotyping Evaluation:
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the
immune cell populations in either the F0 adults rats analysed following administration of Omnirad 184
by oral gavage. - Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- Liver weights in animals dosed at ≥ 100 mg/kg/day, were statistically significantly higher than the
controls and showed a dose-related response. In males, liver weights were up to ~35% higher and in f
emales up to ~28% higher (relative to body weight at 900 mg/kg/day). This was considered test item-r
elated.
Kidney weights in animals dosed at ≥ 100 mg/kg/day, were statistically significantly higher than the c
ontrols and showed a dose-related response. In males, kidney weights were up to ~30% higher and
in females up to ~21% higher (relative to body weight at 900 mg/kg/day). Although this could not be
correlated to any microscopic findings, it was considered test item-related in view of the similar weigh
t differences recorded in the F1 cohort 1A and 1B animals.
Additional Organ Weight Difference:
Testis weights (absolute, relative to %body or %brain weights) were statistically significantly higher by ~12% (relative to body weight at 900 mg/kg/day) compared to the controls. Although the fact that males dosed at 900 mg/kg/day had a lower terminal body weight would increase testis weight relative t o body weight, it is important to note that absolute weight was increased also. Although similar weight differences were seen in the F1 cohort 1A and 1B animals. This was however considered incidental
in view of the minor weight difference and no microscopic findings that could be correlated.
Heart weights were statistically higher in females dosed at ≥ 100 mg/kg/day. No microscopic find
ings could be correlated to this, it was not dose-related, and this difference was not seen in the other
gene rations and it was therefore considered not test item-related.
There were individual organ weight values that were different from their respective controls. There
were, however, no patterns or correlating data to suggest these values were test item-related. - Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test item-related gross findings.
Gross findings observed were of the nature commonly observed in this strain and age of rat, or occur
red at a similar incidence in control and treated animals, and, therefore, were considered not to be
test item-related. - Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Test item-related Microscopic Findings
In the liver, there was diffuse centrilobular hepatocyte hypertrophy of minimal to mild severity. For
males only, this was seen in all dosage groups, whereas for females this was only identified in ani
mals dosed at 900 mg/kg/day. Although this was also found in one control animal (M1021), the
incidence was higher in animals treated with the test material and followed a dose-related distribution.
This finding was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in f
emales dosed at 900 mg/kg/day.
The higher liver weights recorded in animals given the test material, were correlated to the histopatholo
gic findings of centrilobular hypertrophy. This correlation was evident in the males dosed at ≥ 100 mg/
kg/day and females dosed at 900 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal
severity. This was characterised by an increase in the number of goblet cells. For males only, this was
seen in at ≥ 300 mg/kg/day, whereas for females this was only identified in animals dosed at 900 mg/
kg/day. Although this was also found in one control animal (M1013), the incidence was convincingly
higher in animals treated with Omnirad 184 at ≥ 300 mg/kg/day and showed a dose-related distribut
ion. This finding was therefore considered test item-related in males dosed at ≥ 300 mg/kg/day and in
females dosed at 900 mg/kg/day.
Additional Microscopic Findings:
In the spleen, there was a slightly higher incidence of increased hematopoietic cells with a minimal
severity, primarily in males given the test material at a doses of 100 mg/kg/day and 900 mg/kg/day. A
slightly higher incidence was seen in females dosed at 900 mg/kg/day only. However, due to the
overall low incidences and because this is a common background finding, it was considered not test i
tem-related.
Other microscopic findings were of the nature commonly observed in this strain and age of rat, or
occurred at a similar incidence in control and treated animals, and, therefore, were considered not to
be test item-related. - Other effects:
- no effects observed
- Number of abortions:
- no effects observed
- Pre- and post-implantation loss:
- no effects observed
- Description (incidence and severity):
- F0 and F1 Generations:
There were no test item-related effects on number of corpora lutea, implant counts, pre- or post-implantation loss or total number of pups born at up to 900 mg/kg/day. - Total litter losses by resorption:
- no effects observed
- Early or late resorptions:
- effects observed, non-treatment-related
- Description (incidence and severity):
- F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early resorption) and 4511F was not pregnant.
- Dead fetuses:
- no effects observed
- Changes in pregnancy duration:
- no effects observed
- Description (incidence and severity):
- There were no test item-related effects on male or female mating performance or fertility, or duration of gestation in females, at up to 900 mg/kg/day.
- Changes in number of pregnant:
- no effects observed
- Description (incidence and severity):
- F0 control 1520F was found not to be pregnant at scheduled necropsy.
F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early resorption) and 4511F was not pregnant.
F0 Animals 1501F, 1502F, 1506F, 1518F, 1521F (control), 2517F (100 mg/kg/day), 350F2 (300 mg/kg/day) and 4523F (900 mg/kg/day) did not have a positive mating sign during pairing and therefore gave birth before their expected GD 21.
The following Cohort 1B females were euthanised on GD 24 as they had failed to produce litters and were found not to be pregnant; 1629F (control), 2624F, 2627F, 2628F and 2637F (100 mg/kg/day) and 3629F (300 mg/kg/day). - Other effects:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Basis for effect level:
- body weight and weight gain
- changes in number of pregnant
- changes in pregnancy duration
- clinical biochemistry
- clinical signs
- dead fetuses
- early or late resorptions
- effects on pregnancy duration
- food consumption and compound intake
- gross pathology
- haematology
- histopathology: neoplastic
- maternal abnormalities
- mortality
- necropsy findings
- number of abortions
- pre and post implantation loss
- total litter losses by resorption
- Key result
- Abnormalities:
- no effects observed
- Fetal body weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 5-11.
For animals in Cohorts 1A and 1B, there was a general trend of slightly lower body weight gain (up to
7.0%) for males at 900 mg/kg/day compared to controls, but no test item related-effects for males at
100 or 300 mg/kg/day or females at ≥100 mg/kg/day during pre-pairing, gestation or lactation.
In Cohort 2A and for Surplus animals, body weight gain was variable and did not follow any trend. - Reduction in number of live offspring:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
Pup numbers and viability on PND 1 to 4 and PND 4 to 21 were comparable to controls at all test item dose levels administered. - Changes in sex ratio:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
The ratio of male to female pups was similar to controls at all test item dose levels administered. - Changes in litter size and weights:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
Pup numbers and viability on PND 1 to 4 and PND 4 to 21 were comparable to controls at all test item dose levels administered. - Anogenital distance of all rodent fetuses:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
There was no effect on anogenital distance or normalised anogenital distance for males or females at up to 900 mg/kg/day. - Changes in postnatal survival:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
Pup numbers and viability on PND 1 to 4 and PND 4 to 21 were comparable to controls at all test item dose levels administered. - External malformations:
- not specified
- Skeletal malformations:
- not specified
- Visceral malformations:
- not specified
- Other effects:
- no effects observed
- Description (incidence and severity):
- Dam and Litter Observations During Lactation:
There were no test item-related effects noted for the F0 or Cohort 1B dams or their litters during lactation at up to 900 mg/kg/day.
Nipple Retention:
F0 and F1 Generations:
On PND 13, there was no effect on nipple retention in male pups i.e. no nipples were present, at dose levels up to 900 mg/kg/day.
Assessment of Sexual Maturation:
F1 Generation:
There was considered to be no effect on sexual maturation for F1 males or females at up to 900 mg/kg/day.
Acoustic Startle:
F1 Generation:
There were no test item-related effects on maximum response amplitude, average response amplitude or time to maximum response in males or females at up to 900 mg/kg/day.
Motor Activity:
F1 Generation:
For males at 900 mg/kg/day, the number of basic and fine movements were lower than controls by up to 70.1% from 40 to 60 minutes of the 1 hour session; this was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this reduction in motor activity, the cause of this apparent reduction in motor activity could not be determined.
At 100 or 300 mg/kg/day for males and ≥ 100 mg/kg/day for females, there were no notable effects on overall basic or fine activity or ambulation, compared with control values.
Brain Morphometry Evaluation:
Cohort 2A
No test item-related effects were seen in the brain morphometry parameters.
The length and width of the brain, as measured grossly, were compared between controls and animals at 900 mg/kg/day. The mean values were similar and were considered not to show a test item-relationship.
Cohort 2B
The only statistically significant difference was that the thickness of the cerebral cortex (motor) in males was ~7% less than controls at 900 mg/kg/day. This was considered due to chance and not test item-related, because other parameters were not affected and in females, the thickness of the cerebral cortex (motor) was not statistically significantly less at 900 mg/kg/day compared to the controls (it was only ~5% less).
Other brain morphometry parameters at 900 mg/kg/day were different from the control means, but were within 10% of the controls and not statistically significant, so were considered not test item-related.
The length and width of the brain, as measured grossly, were compared between controls and animals at 900 mg/kg/day. The mean values were similar and were considered not to show a test item-relationship. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- reduction in number of live offspring
- changes in sex ratio
- fetal/pup body weight changes
- changes in litter size and weights
- changes in postnatal survival
- external malformations
- skeletal malformations
- visceral malformations
- Key result
- Abnormalities:
- no effects observed
- Developmental effects observed:
- no
- Conclusions:
- The administration of the target substance once daily (or twice daily 3 hours apart on some occasions) via
oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no
test item-related deaths. There were occasional observations of decreased activity, abnormal, un
coordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose le
vel-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body we
ight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/
day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. Ther
e were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and
F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity
was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirme
d by overall ambulation values, or any associated clinical findings, detailed functional observations or
brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/
kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 male
s only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels
(up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values
or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target
organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertro
phy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney
weights were seen in both sexes. In neurological investigations, there were no test item-related brain
weight differences, gross or histological findings or effects on the brain dimensions. There were no
histological findings in the reproductive tissues of the females that had not been observed to litter tha
t would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1
adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the i
n-life and pathological findings were considered not to be adverse. For F1 and F2 developmental para
meters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on
the pup numbers, survival or development. - Executive summary:
Read-across approach:
The methodology used for choosing potential read-across source substances for this current assessment was two-fold:
- Identify substances that have a similar molecular structure to the target substance, for example, having common functional groups as well as likely having structurally-similar degradation products, and
- Identify which of these source substances have the necessary toxicology data to fulfil the data gap.
The substance finally selected for the read-across source met the scientific hypothesis and comparison criteria below:
- The target and source substance are small molecules with common functional groups which will influence their toxicokinetic behaviour. The predicted metabolic route and hence breakdown products within the body are common for both substances.
- A comparison of existing mammalian (rat) toxicity data shows that both substances were of low toxicity, with both having high values for No Observed Adverse Effect Levels (NOAEL) for repeated dose toxicity studies in rats. In particular, no developmental toxicity was observed in either substance at the dose levels tested.
- The intended use of both substances is as Photoinitiators which means high commonality in the manufacture, formulation and use. Thus the potential for (accidental) human and environmental exposure is the same for both substances.
It is therefore concluded that the target substance will have the same reproductive toxicity outcomes as that of the source substance for an Extended One Generation Reproductive Toxicity (EOGRT) Study.
Executive summary of the study:
The objective of this study was to determine the potential toxicity of the test item, when given by oral gavage to adult rats and their offspring. This study was designed to provide an evaluation of reproduction and development as well as a thorough evaluation of systemic toxicity in pregnant and lactating females and their offspring. Detailed examination of key developmental endpoints, such as offspring viability, neonatal health, developmental status at birth, physical and functional development until adulthood, nervous and immune system development was expected to identify any specific target organs in the offspring. In addition, the study provided information about the effects of the test item on the integrity and performance of the adult male and female reproductive systems and a neurobehavioural assessment of the F1 generation.
The study design was as follows:
Text Table 1 Experimental Design – F0 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
25
25
2
Test item
100
10
10
10.03
100
25
25
3
Test item
300
10
30
30.09
301
25
25
4
Test item
900
10
90
90.27
903
25
25
a Dose volume was based on the most recent body weight measurement.
Control/vehicle = 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 2 Experimental Design – F1 Unselected PND (Post Natal Day) 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 3 Experimental Design – Cohort 1A (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Test item
100
10
10
10.03
100
20
20
3
Test item 300
10
30
30.09
301
20
21d
4
Test item
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and
450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
d One female given 300 mg/kg/day died within minutes of receiving its first dose on Day 1, and another female
was added to the group.Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 4 Experimental Design – Surplus F1 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Test item
100
10
10
10.03
100
10
10
3
Test item
300
10
30
30.09
301
10
10
4
Test item
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 5 Experimental Design – Cohort 1B (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Test item
100
10
10
10.03
100
20
20
3
Test item
300
10
30
30.09
301
20
20
4
Test item
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 6 Experimental Design – F2 Unselected PND 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 7 Experimental Design – Cohort 2A (Neurobehavioural)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Test item
100
10
10
10.03
100
10
10
3
Test item
300
10
30
30.09
301
10
10
4
Test item
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 8 Experimental Design – Cohort 2B (Neuropathology)a
Group No.
Number of Animals
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Cohort 2B were not dosed as they were terminated on PND 21/22.
In Week 17 of the study (F0 dosing and the start of F1 dosing), the 900 mg/kg/day dosing formulation could not be pulled up into the gavage tube being used for newly weaned pups, due to an unknown viscosity property, resulting in some subsets of F0 and/or F1 animals either not being dosed or being given the 300 mg/kg/day formulation either once or twice daily instead (refer to section 4.6.1 for details). From 19 Aug 2021, Group 4 F1 animals were dosed twice daily 3 hours apart at 450 mg/kg (equivalent to 900 mg/kg/day) until they were old enough for the next size gavage tube on 30 Aug 2021. Although there were occasions when animals were not dosed or received a dose level lower than that required by the study protocol, there was considered to be no overall impact on the results or study outcome due to the length of time that each generation was administered the test item dose levels correctly.
The following parameters and end points were evaluated in this study: clinical observations, body weights, food consumption, estrous cycles, mating performance, fertility indices, duration of gestation and overall litter performance, litter survival indices, litter and pup weights, pre-weaning physical development of F1 pups, assessments of sexual maturation of F1 animals, clinical pathology parameters (haematology, coagulation, clinical chemistry, and urinalysis), thyroid stimulating hormone (TSH) and thyroxine (T4) analysis, gross necropsy findings, splenic immunophenotyping analysis, organ weights, sperm evaluation, ovarian follicle counts and histopathological examinations.
None of the unscheduled deaths in any generation were considered to be test item-related.
For the F0 generation at 900 mg/kg/day, test item-related clinical observations of decreased activity, abnormal/uncoordinated gait and low carriage were occasionally seen for males and/or females before pairing, and decreased activity only during lactation. For F0 and F1 generations, salivation and/or ploughing were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, throughout the dosing periods. F0 and F1 males given 900 mg/kg/day had slightly lower body weight gain, and F0 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day.
There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development.
For males at 900 mg/kg/day, during the last 20 minutes of the 1 hour session, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this, the cause of this apparent reduction in motor activity could not be determined.
For F0 males given ≥300 mg/kg/day and F0 females given ≥100 mg/kg/day, triglycerides were lower by up to 0.75-fold, and for males only, cholesterol higher by up to 1.3-fold. For F1 animals at ≥100 mg/kg/day, triglycerides were lower than controls by up to 0.62-fold, and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold. There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day.
In the F0 animals dosed from 10 weeks prior to mating and continuing throughout mating and gestation until the females were at least LD 21, microscopically, centrilobular hypertrophy in the liver was considered test item-related in all dose groups in males but only at 900 mg/kg/day in females and mucosal hyperplasia of the glandular region, appearing as an increased number of goblet cells, was considered test item-related at ≥ 300 mg/kg/day in males and only at 900 mg/kg/day in females. Higher liver and kidney weights were seen in both sexes at all dose levels.
In the F1 cohort 1A animals, dosed from PND 21 until at least PND 90, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. In the stomach, there was mucosal hyperplasia of the glandular regions, appearing as an increased number of goblet cells. This finding was considered test item-related in males and females at ≥ 300 mg/kg/day. Liver weights were higher at ≥ 300 mg/kg/day in males and females. In addition, higher kidney weights were seen in females at all dose levels and at ≥ 300 mg/kg/day in males.
In the F1 cohort 1B animals, that were used to produce an F2 generation, and that were dosed from PND 21 until at least LD 21, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. Liver weights were higher at all dose levels in males and only at 900 mg/kg/day in females. In addition, higher kidney weights were seen in animals dosed at 900 mg/kg/day. Stomachs were not assessed microscopically in these animals. There were no test item-related organ weight differences in the female reproductive tissues.
There were no test item-related organ weight differences in a limited list of tissues in F1 and F2 pups that were euthanised at PND 21 without active dosing.
In the F1 cohort 2A and 2B animals used for neurological investigations (euthanised on PND 21 or dosed from PND 21 until at least PND 78-80), there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions.
There were no test item-related gross findings in the entire study, and there were only 6 premature deaths, all of which were considered not test item-related.
There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility.
In conclusion, administration of the test item by once daily (or twice daily 3 hours apart on some occasions) oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no test item-related deaths. There were occasional observations of decreased activity, abnormal, uncoordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose level-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body weight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirmed by overall ambulation values, or any associated clinical findings, detailed functional observations or brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 males only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels (up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertrophy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney weights were seen in both sexes. In neurological investigations, there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions. There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1 adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the in-life and pathological findings were considered not to be adverse. For F1 and F2 developmental parameters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on the pup numbers, survival or development.
- Endpoint:
- developmental toxicity
- Remarks:
- extended one-generation reproductive toxicity - with F2 generation and developmental neurotoxicity (Cohorts 1A, 1B with extension, 2A and 2B)
- Type of information:
- experimental study
- Remarks:
- Performed on source substance.
- Adequacy of study:
- key study
- Study period:
- April 2021 - April 2022
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 424. Neurotoxicity Study in Rodents
- Version / remarks:
- 21 July 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 443 Extended One-Generation Reproductive Toxicity Study
- Version / remarks:
- 25 June 2018
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Specific details on test material used for the study:
- - Identification: Omnirad 184
- Alternate identification: Hydroxycyclohexyl phenyl ketone
- EC number: 213-426-9
- CAS number: 947-19-3
- Physical description: Light white solid
- Storage condition of test material: Ambient, protected from light
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Test Item is stable in the vehicle when prepared and stored under the same conditions at concentrations bracketing those used in the present study - Species:
- rat
- Strain:
- Wistar
- Remarks:
- Han Wistar (Crl:WI(Han))
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River UK, Margate, Kent, UK
- Age at study initiation: ca 6-7 weeks for males and 5-6 weeks for females
- Weight at study initiation: 150 to 250 g (males)/100 to 150 g (females)
- Housing: Group housed (up to 3 or 4 animals of the same sex and same dosing group together). A
few days prior to mating, F0 and Cohort 1B males were be transferred to individual cages with solid
bottoms. F0 and Cohort 1B females were transferred to these cages for mating. Mated F0 and Co
hort 1B females were transferred to individual solid bottomed cages. White paper tissue was supplied
as nesting material from Gestation Day 20. F0 and Cohort 1B females with litters were retained in t
his type of cage until termination. On a suitable day after completion of mating, the F0 and Cohort 1B
males were re-housed with their original cage mates.
- Diet (e.g. ad libitum): Special Diet Services VRF1. Ad libitum, except during designated procedures
- Water (e.g. ad libitum): Public supply tap water. Freely available to each animal from water bottles
(except during
designated procedures).
- Acclimation period: The F0 animals were allowed to acclimate to the Test Facility rodent toxicology
accommodation for 10 days before the commencement of dosing (replacement animals at least 5
days).
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23°C
- Humidity (%): 40 - 70%
- Air changes (per hr): Ten or more air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark (except during designated pro
cedures)
IN-LIFE DATES: From: 12 Apr 2021 To: 21 Dec 2021 - Route of administration:
- oral: gavage
- Vehicle:
- CMC (carboxymethyl cellulose)
- Remarks:
- CMC (carboxymethyl cellulose) CMC (carboxymethyl cellulose) 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q Water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: Dose formulations divided into aliquots where required to
allow them to be dispensed on each dosing occasion
VEHICLE
- Justification for use and choice of vehicle: Carboxymethylcellulose, analysis performed in a s
eparate study showed good stability in this vehicle
- Concentration in vehicle: 10, 30, 90mg/mL
- Amount of vehicle: 10ml/kg - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Analyses was performed by Gas Chromatography (GC) using a validated analytical procedure.
Concentration and Homogeneity Analysis
Sample Allocation: 2 for analysis, 3 for backup for control; 2 for analysis and 3 for backup per stratum for Groups 2-4.
Sampling Containers: Appropriate sized volumetric flasks
Storage Conditions: Temperature set to maintain 4°C, protected from light
Acceptance Criteria: For concentration: mean sample concentration results within or equal to
± 15% of theoretical concentration. Each individual sample concentration result within or equal to ±
20%. For homogeneity, relative standard deviation (RSD) of concentrations of ≤ 10% for each group.
Stability analyses performed previously in conjunction with 426730 demonstrated that the Test Item
is stable in the vehicle when prepared and stored under the same conditions at concentrations
bracketing those used in the present study. - Details on mating procedure:
- Frequency: F0 - Day 71
Cohort 1B – PND 107-111
Procedure: Pairing was on a 1 male to 1 female basis. During the evening (after 5 pm), females
were housed with their allocated co-group male partner. The animals were paired in ascending numer
ical order. Vaginal lavages were taken early each morning from the day of pairing until mating
had occurred and the stage of estrous observed in each vaginal lavage was recorded. The day of
detection of a copulatory plug in situ and/or of sperm in the lavage was designated Gestation Day 0.
The pairing period for each pair of animals was a maximum of 14 nights (unless a longer period was
deemed appropriate).
If evidence of mating was not observed by the end of the pairing period, the female was separated f
rom the male during the morning following the last night of pairing and treated as if mating had occ
urred during that night. Procedures for that female were continued as if it had mated on the last
night of pairing.
For each female the time taken to show a positive mating sign and the number of failed opportunities t
o mate (estruses passed without a sign of mating) was evaluated. - Duration of treatment / exposure:
- F0 Males: 10 weeks prior to mating and continuing throughout and after mating until the day before t
ermination.
F0 Females: 10 weeks prior to mating and continuing throughout mating and gestation until at least
LD 21.
Cohort 1A: From PND 21 until the day before necropsy
Surplus Animals: From PND 21 until the day before necropsy
Cohort 1B: From PND 21 until the day before necropsy
Cohort 2A: From PND 21 until the day before necropsy - Frequency of treatment:
- Once daily.
From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at
10mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day. - Duration of test:
- F0 Males: 10 weeks prior to mating and continuing throughout and after mating until the day before t
ermination.
F0 Females: 10 weeks prior to mating and continuing throughout mating and gestation until at least
LD 21.
Cohort 1A: From PND 21 until the day before necropsy
Surplus Animals: From PND 21 until the day before necropsy
Cohort 1B: From PND 21 until the day before necropsy
Cohort 2A: From PND 21 until the day before necropsy - Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- control
- Dose / conc.:
- 100 mg/kg bw/day
- Dose / conc.:
- 300 mg/kg bw/day
- Dose / conc.:
- 900 mg/kg bw/day
- No. of animals per sex per dose:
- 10 animals per sex per dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- Dose selection rationale:
- F0 animals were randomly assigned to groups. Males and females were randomised separately.
- Fasting period before blood sampling for clinical biochemistry: animals not fasted - Maternal examinations:
- F0 Animals
CAGE SIDE OBSERVATIONS:
Pre and Postdose Observations, All F0 animals.
Frequency: Once predose and up to 4 hours postdose after the end of each group for each sex.
DETAILED CLINICAL OBSERVATIONS:
All F0 animals Weekly; from at least Week 1 and throughout the study.
BODY WEIGHT:
All F0 animals.
Males: weekly beginning Week -1.
Females: weekly beginning Week -1 until pairing for mating and then on GD 0, 7, 14 and 20 and LD 1,
4, 7, 14 and 21.
FOOD CONSUMPTION:
All F0 animals.
Males: weekly from Week -1 until pairing for mating. Then weekly after mating and re-housing.
Females: weekly beginning week -1 until pairing for mating and then on GD 0-7, 7-14 and 14-20, and
LD 1-7, 7-14 and 14-21
WATER CONSUMPTION:
All F0 animals. Regular basis throughout the study.
Mortality:
All F0 animals. At least twice daily (once at the start and once towards the end of the working day)
beginning upon arrival through termination/release. - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination
Examinations included:
- Gravid uterus weight
- Number of corpora lutea
- Number of implantations
- Number of early resorptions
- Number of late resorptions - Blood sampling:
- Blood sampling for TSH and T4 analysis
- Fetal examinations:
- The numbers of live and dead pups born in each litter were recorded as soon as possible after
completion of parturition on LD 0. The live pups were counted and examined daily for the presence o
f milk in the stomach and for any externally visible abnormalities up to and including PND 4 and then
on PND 7, 14 and 21. Individual pup weights were taken for each litter on PND 1, 4, 7, 14 and 21. A
head count of the pups was performed each day.
Ano-genital distance was measured for the pups of both sexes on PND 1. Nipple retention was
assessed in males on PND 13.
Assessment of Sexual Maturation Cohorts 1A+ Surplus Animals, 1B and 2A:
Commencing on PND 28, females were examined daily for vaginal opening. The day on which the va
gina became open was recorded, as was the body weight on that day.
Commencing on PND 35, males were examined daily for balanopreputial separation. The day on wh
ich separation occurred was recorded, as was the body weight on that day.
GROSS EXAMINATION OF DEAD PUPS:
Pups Found Dead or Euthanised Prematurely Before PND 14: Where practicable, these animals were
sexed and then checked for the presence of milk in the stomach and for the presence of any extern
ally visible abnormalities. Any externally abnormal pups were fixed in 10% neutral buffered formalin
for optional further examination. Externally normal pups were discarded.
Pups Found Dead or Euthanised Prematurely On or After PND 14: These animals were subject to a
gross necropsy. An external examination was followed by an inspection of the cranial, thoracic and
abdominal contents. Internal sex was also confirmed. Representative samples of any abnormal t
issues were taken and fixed in neutral buffered 10% neutral buffered formalin. These carcasses were
then discarded.
ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:
Perfusion fixation, neuropathology and brain morphometry was performed for all animals. The fixed b
rains were removed and weighed, and the length and maximum width of the brain was measured for
all animals selected for neuropathology.
Brain, dorsal root ganglion, and spinal cords tissues were prepared for neuropathology examination.
Neuropathology and brain morphometry were performed.
ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:
Lymph nodes were weighed to evaluate immunotoxic effects from 10 rats/sex/group from Groups 1-4.
One-half of the spleen was used for immunophenotyping assessment from 10 rats/sex/group from
Groups 1-4.
Necropsy (F1 and F2 Culled PND 4 and Unselected PND 21 pups):
Culled pups on PND 4 were necropsied. Necropsy consisted of external examination followed by
macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities
in situ. Samples of any grossly abnormal tissues were preserved in 10% neutral buffered formalin or
other appropriate fixative. The carcasses were discarded.
All unselected pups on PND 21 were necropsied. Necropsy consisted of external examination
followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdomina
l cavities in situ. Representative samples of abnormal tissues were preserved in 10% neutral buffere
d formalin or other appropriate fixative. For F1 and F2 pups, tissue samples and weights of the foll
owing tissues were taken from 10 male and 10 female pups per group and were preserved in 10%
neutral buffered formalin or other appropriate fixative: Brain, mammary gland, parathyroid gland,
thyroid gland, liver, ovary, spleen, testes, thymus.
Where 10 pups of each sex were not available for organ weight and tissue collection, additional pups
of the opposite sex were selected such that 20 animals per group had organ weights and tissues
collected.
Necropsy (Cohort 1B Animals):
Animals were subjected to a complete necropsy examination, which included evaluation of the carcas
s and musculoskeletal system; all external surfaces and orifices; cranial cavity and external surfaces
of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and tissues.
The reproductive tracts of all F1 females were examined for signs of implantation, the number of any
implantation sites being recorded. The total number of corpora lutea graviditatis were recorded for
each female. For F1 females necropsied on GD 24 as they had failed to produce a litter, the uteri
of all non-pregnant females were fixed in 10% neutral buffered formalin after being examined for
implantation sites. On GD 24, if a female was found to be pregnant, the number and type of any impl
antation sites were recorded and the total number of corpora lutea graviditatis were recorded also.
Transcardial Perfusion Fixation of Cohorts 2A and 2B:
All animals were subjected to a limited examination, with special attention being paid to the reproducti
ve organs.
The head (with brain exposed), spinal cord and hindlimbs (Cohort 2A) and head (with brain exposed)
(Cohort 2B) was placed in 10% neutral buffered formalin and allowed to fix for at least 7 days. The
fixed brains were removed and weighed, and the length and maximum width of the brain was
measured for all animals selected for neuropathology. Subsequently, the brain was fixed in 10% neut
ral buffered formalin together with selected PNS tissues. - Statistics:
- Body Weight Changes: Calculated between each scheduled interval
Food Consumption: Calculated between appropriate intervals
Organ Weight Relative to Body Weight: Calculated against the terminal body weight for scheduled
intervals
Organ Weight Relative to Brain Weight: Calculated against the brain weight for scheduled intervals
Descriptive Statistical Analyses:
Means, standard deviations, percentages, numbers, and/or incidences have been reported as approp
riate by dataset.
Inferential Statistical Methods:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were c
onducted using two sided tests and have been reported at the 1% and 5% levels.
The pairwise comparisons of interest are listed below:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1
Analyses were performed according to the matrix below when possible but excluded any group with
less than 3 observations.
See Table 43 below.
Parametric/Non-parametric:
Levene’s test was used to assess the homogeneity of group variances.
The groups were compared using an overall one-way ANOVA F-test if Levene’s test was not signif
icant or the Kruskal-Wallis test if it was significant. If the overall F-test or Kruskal Wallis test was found
to be significant, then pairwise comparisons were conducted using Dunnett’s or Dunn’s test, respect
ively.
Non-Parametric:
The groups were compared using an overall Kruskal-Wallis test. If the overall Kruskal-Wallis test
was found to be significant, then the above pairwise comparisons were conducted using Dunn’s test
(equivalent to Wilcoxon Rank-Sum test in Nevis 2012 tables). - Indices:
- Reproductive indices
Female Mating Index = Number of Females with Evidence of Mating (or no confirmed mating date and
pregnant) / Number of Females Paired
Female Fertility Index = Number of Pregnant Females / Number of Females with Evidence of Mating
(or no confirmed mating date and pregnant)
Female Pregnancy Index = Number of Pregnant Females / Number of Females Paired
Male Mating Index = Number of Males with Evidence of Mating (or female partner confirmed pregna
nt) / Number of Males Paired
Male Fertility Index = Number of Males Impregnating a Female / Number of Males with Evidence of
Mating (or female partner confirmed pregnant)
Male Pregnancy Index = Number of Males Impregnating a Female / Number of Males Paired
The following natural delivery/reproductive parameters were included, as appropriate:
Gestation Length: The gestation length was calculated from GD 0 to the day the first pup was
observed.
Gestation Index:
Percentage of pregnancies that result in birth of live litters = (Number of Animals with Live Offspring /
Number of Animals Pregnant) x 100
Offspring viability indices
Live Birth Index:
Percentage of pups born alive = (Number of Live Newborn Pups / Number of Newborn
Pups) x100
Sex Ratio (% males):
Percentage of male pups per litter = (Number of Live Male Pups / Total Number of Live Pups) x100
Viability Index:
Percentage of pups born that survive 4 days postpartum = (Number of Live Pups on Day 4 Postpa
rtum / Number of Live Newborn Pups) x100
Lactation Index:
Percentage of pups that survive 21 days postpartum = (Number of Live Pups on Day 21 Postpartum /
Number of Live Pups on Day 4 (postculling) Postpartum) x100
Post-Implantation Loss/Litter = (Number of Implants – Total Newborn Pups / Number of Implants)
x100 - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- At 900 mg/kg/day, decreased activity was observed for one male on Day 2 and abnormal, unco
ordinated gait for 3 males and 4 females between Days 2 to 4. Decreased activity, abnormal, uncoord
inated gait or low carriage were occasionally noted for a further 8 females between Days 5 to 70 and
decreased activity on one occasion for each of 2 females during lactation. These findings were cons
idered to be test item-related.
Salivation and/or ploughing (also a chewing action for one female at 900 mg/kg/day) were noted for
all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, on multiple days thro
ughout the dosing period. The incidence of these findings was dose level-related.
Animal 4515F (900 mg/kg/day) had abnormal clincal observations of irregular respiratory rate, hunc
hed posture, erect fur, cold to touch, prominent backbone, abnormal gait, decreased activity and red
discharge from vagina the day after parturition (LD 1) but these mostly resolved before scheduled
termination. The clinical observations were considered to be likely due to the trauma of giving birth
rather than being test item-related.
Animal 4525F (900 mg/kg/day) had abnormal clincal observations of irregular/decreased respiratory
rate, erect fur, eyes partly closed, decreased activity up to 2-4 hours postdose on LD 7 only. No
similar findings were observed for this animal during the dosing period and as a cause could not be determined, these findings were likely related to the dosing procedure. - Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- None of the unscheduled deaths were considered to be test item-related.
On Study Day 40, approximately 3 hours after dosing, Animal 4510F given 900 mg/kg/day was found
with clinical signs of irregular respiratory rate, hunched posture, partly closed eyes, erect fur, unc
oordinated and abnormal gait and decreased activity and was euthanised. There were no findings
at necropsy or microscopically to suggest the likely cause of death. This animal had not had any
abnormal clinical signs before this (apart from salivation and ploughing) and had consistently gained
body weight.
Animals 2512F and 4502F given 100 or 900 mg/kg/day respectively, were euthanised on LD 1 or LD
0 and due to the very young age of the pups, they were also euthanised. Animal 2512F had clinical
signs of irregular respiratory rate, hunched posture, pale skin and cold to touch, erect fur, uncoor
dinated and abnormal gait in the afternoon of LD 1. Animal 4502F was found subdued, cold to touch,
prostrate/lying on side with decreased respiratory rate approximately 1 hour after dosing on LD 0. Th
ere were no findings at necropsy or microscopically to suggest the likely cause of death and both an
imals had not had any abnormal clinical signs before this (apart from salivation and ploughing for 4
502F) and had consistently gained body weight. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See Tables 1-4.
For males given 900 mg/kg/day, there was slightly lower body weight gain over the dosing period
resulting in male body weight being 3.6% lower than the concurrent control mean value by Day 134.
For females given ≥ 100 mg/kg/day, a dose level-related higher body weight gain of up to 10.3% at 90
0 mg/kg/day compared to controls was noted during the pre-pairing period and this slightly higher bo
dy weight was generally maintained throughout gestation and lactation.
There was no test item-related effect on male body weight at 100 or 300 mg/kg/day. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- There were occasional slightly higher food consumption values of up to 14.4% of controls noted for
males at 300 or 900 mg/kg/day up to Day 106. At ≥100 mg/kg/day for males from Days 106 to 134,
there was higher food consumption of up to 17.5%. For females given ≥100 mg/kg/day, there was an
overall dose level-related higher food consumption of up to 20.6% of control values during the prep
airing
period, and during gestation and lactation, food consumption was higher for females by up to
13.2% at 300 or 900 mg/kg/day only. - Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Haematology
See tables 18-19.
Coagulation
See tables 15-16.
All differences in haematology or coagulation parameters were considered not to be test item-related
based on their small magnitude, inconsistent direction, absence of a dose response, general overl
ap of individual values with the range of control values and/or were of a magnitude of variation com
monly observed in rats. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 12-13.
At ≥300 mg/kg/day for males and ≥100 mg/kg/day for females, triglycerides were lower than controls
by up to 0.75-fold, and for males only at ≥300 mg/kg/day, cholesterol higher by up to 1.30-fold. - Endocrine findings:
- effects observed, treatment-related
- Description (incidence and severity):
- There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of
necropsy in F0 adult males given 300 or 900 mg/kg/day.
There were no test item-related effects for F0 adult males given 100 mg/kg/day or F0 adult females gi
ven up to 900 mg/kg/day or for culled pups on PND 4 or unselected pups on PND 21.
There were no test item-related changes to T4 levels at up to 900 mg/kg/day for samples taken on the
morning of necropsy for the F0 generation animals, for culled pups on PND 4 or unselected pups on
PND 21.
The differences in these parameters were considered not to be test item-related based on their small
magnitude, inconsistent direction, absence of a dose response, general overlap of individual values
with the range of control and/or were of a magnitude of variation commonly observed in rats. - Urinalysis findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- See tables 27-28.
All differences in urine parameters were considered not to be test item-related based on their small m
agnitude, inconsistent direction, absence of a dose response, general overlap of individual values with
the range of control and/or were of a magnitude of variation commonly observed in rats. - Immunological findings:
- no effects observed
- Description (incidence and severity):
- Splenic Immunophenotyping Evaluation:
The results demonstrated no test item, dose-dependent or sex-dependent effects on any of the
immune cell populations in either the F0 adults rats analysed following administration of Omnirad 184
by oral gavage. - Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- Liver weights in animals dosed at ≥ 100 mg/kg/day, were statistically significantly higher than the
controls and showed a dose-related response. In males, liver weights were up to ~35% higher and in f
emales up to ~28% higher (relative to body weight at 900 mg/kg/day). This was considered test item-r
elated.
Kidney weights in animals dosed at ≥ 100 mg/kg/day, were statistically significantly higher than the c
ontrols and showed a dose-related response. In males, kidney weights were up to ~30% higher and
in females up to ~21% higher (relative to body weight at 900 mg/kg/day). Although this could not be
correlated to any microscopic findings, it was considered test item-related in view of the similar weigh
t differences recorded in the F1 cohort 1A and 1B animals.
Additional Organ Weight Difference:
Testis weights (absolute, relative to %body or %brain weights) were statistically significantly higher by ~12% (relative to body weight at 900 mg/kg/day) compared to the controls. Although the fact that males dosed at 900 mg/kg/day had a lower terminal body weight would increase testis weight relative t o body weight, it is important to note that absolute weight was increased also. Although similar weight differences were seen in the F1 cohort 1A and 1B animals. This was however considered incidental
in view of the minor weight difference and no microscopic findings that could be correlated.
Heart weights were statistically higher in females dosed at ≥ 100 mg/kg/day. No microscopic find
ings could be correlated to this, it was not dose-related, and this difference was not seen in the other
gene rations and it was therefore considered not test item-related.
There were individual organ weight values that were different from their respective controls. There
were, however, no patterns or correlating data to suggest these values were test item-related. - Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test item-related gross findings.
Gross findings observed were of the nature commonly observed in this strain and age of rat, or occur
red at a similar incidence in control and treated animals, and, therefore, were considered not to be
test item-related. - Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Test item-related Microscopic Findings
In the liver, there was diffuse centrilobular hepatocyte hypertrophy of minimal to mild severity. For
males only, this was seen in all dosage groups, whereas for females this was only identified in ani
mals dosed at 900 mg/kg/day. Although this was also found in one control animal (M1021), the
incidence was higher in animals treated with Omnirad 184 and followed a dose-related distribution.
This finding was therefore considered test item-related in males dosed at ≥ 100 mg/kg/day and in f
emales dosed at 900 mg/kg/day.
The higher liver weights recorded in animals given Omnirad 184, were correlated to the histopatholo
gic findings of centrilobular hypertrophy. This correlation was evident in the males dosed at ≥ 100 mg/
kg/day and females dosed at 900 mg/kg/day.
In the glandular portion of the stomach, there was diffuse mucosal hyperplasia with a minimal
severity. This was characterised by an increase in the number of goblet cells. For males only, this was
seen in at ≥ 300 mg/kg/day, whereas for females this was only identified in animals dosed at 900 mg/
kg/day. Although this was also found in one control animal (M1013), the incidence was convincingly
higher in animals treated with Omnirad 184 at ≥ 300 mg/kg/day and showed a dose-related distribut
ion. This finding was therefore considered test item-related in males dosed at ≥ 300 mg/kg/day and in
females dosed at 900 mg/kg/day.
Additional Microscopic Findings:
In the spleen, there was a slightly higher incidence of increased hematopoietic cells with a minimal
severity, primarily in males given Omnirad 184 at a doses of 100 mg/kg/day and 900 mg/kg/day. A
slightly higher incidence was seen in females dosed at 900 mg/kg/day only. However, due to the
overall low incidences and because this is a common background finding, it was considered not test i
tem-related.
Other microscopic findings were of the nature commonly observed in this strain and age of rat, or
occurred at a similar incidence in control and treated animals, and, therefore, were considered not to
be test item-related. - Other effects:
- no effects observed
- Number of abortions:
- no effects observed
- Pre- and post-implantation loss:
- no effects observed
- Description (incidence and severity):
- F0 and F1 Generations:
There were no test item-related effects on number of corpora lutea, implant counts, pre- or post-implantation loss or total number of pups born at up to 900 mg/kg/day. - Total litter losses by resorption:
- no effects observed
- Early or late resorptions:
- effects observed, non-treatment-related
- Description (incidence and severity):
- F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early resorption) and 4511F was not pregnant.
- Dead fetuses:
- no effects observed
- Changes in pregnancy duration:
- no effects observed
- Description (incidence and severity):
- There were no test item-related effects on male or female mating performance or fertility, or duration of gestation in females, at up to 900 mg/kg/day.
- Changes in number of pregnant:
- no effects observed
- Description (incidence and severity):
- F0 control 1520F was found not to be pregnant at scheduled necropsy.
F0 Control Animal 1524F and Animal 4511F given 900 mg/kg/day were terminated on GD 24 as they had failed to produce litters. At necropsy, 1524F had 2 implants (one early resorption) and 4511F was not pregnant.
F0 Animals 1501F, 1502F, 1506F, 1518F, 1521F (control), 2517F (100 mg/kg/day), 350F2 (300 mg/kg/day) and 4523F (900 mg/kg/day) did not have a positive mating sign during pairing and therefore gave birth before their expected GD 21.
The following Cohort 1B females were euthanised on GD 24 as they had failed to produce litters and were found not to be pregnant; 1629F (control), 2624F, 2627F, 2628F and 2637F (100 mg/kg/day) and 3629F (300 mg/kg/day). - Other effects:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Basis for effect level:
- body weight and weight gain
- changes in number of pregnant
- changes in pregnancy duration
- clinical biochemistry
- clinical signs
- dead fetuses
- early or late resorptions
- effects on pregnancy duration
- food consumption and compound intake
- gross pathology
- haematology
- histopathology: neoplastic
- maternal abnormalities
- mortality
- necropsy findings
- number of abortions
- pre and post implantation loss
- total litter losses by resorption
- Key result
- Abnormalities:
- no effects observed
- Fetal body weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- See tables 5-11.
For animals in Cohorts 1A and 1B, there was a general trend of slightly lower body weight gain (up to
7.0%) for males at 900 mg/kg/day compared to controls, but no test item related-effects for males at
100 or 300 mg/kg/day or females at ≥100 mg/kg/day during pre-pairing, gestation or lactation.
In Cohort 2A and for Surplus animals, body weight gain was variable and did not follow any trend. - Reduction in number of live offspring:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
Pup numbers and viability on PND 1 to 4 and PND 4 to 21 were comparable to controls at all test item dose levels administered. - Changes in sex ratio:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
The ratio of male to female pups was similar to controls at all test item dose levels administered. - Changes in litter size and weights:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
Pup numbers and viability on PND 1 to 4 and PND 4 to 21 were comparable to controls at all test item dose levels administered. - Anogenital distance of all rodent fetuses:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
There was no effect on anogenital distance or normalised anogenital distance for males or females at up to 900 mg/kg/day. - Changes in postnatal survival:
- no effects observed
- Description (incidence and severity):
- F1 and F2 Generations:
Pup numbers and viability on PND 1 to 4 and PND 4 to 21 were comparable to controls at all test item dose levels administered. - External malformations:
- not specified
- Skeletal malformations:
- not specified
- Visceral malformations:
- not specified
- Other effects:
- no effects observed
- Description (incidence and severity):
- Dam and Litter Observations During Lactation:
There were no test item-related effects noted for the F0 or Cohort 1B dams or their litters during lactation at up to 900 mg/kg/day.
Nipple Retention:
F0 and F1 Generations:
On PND 13, there was no effect on nipple retention in male pups i.e. no nipples were present, at dose levels up to 900 mg/kg/day.
Assessment of Sexual Maturation:
F1 Generation:
There was considered to be no effect on sexual maturation for F1 males or females at up to 900 mg/kg/day.
Acoustic Startle:
F1 Generation:
There were no test item-related effects on maximum response amplitude, average response amplitude or time to maximum response in males or females at up to 900 mg/kg/day.
Motor Activity:
F1 Generation:
For males at 900 mg/kg/day, the number of basic and fine movements were lower than controls by up to 70.1% from 40 to 60 minutes of the 1 hour session; this was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this reduction in motor activity, the cause of this apparent reduction in motor activity could not be determined.
At 100 or 300 mg/kg/day for males and ≥ 100 mg/kg/day for females, there were no notable effects on overall basic or fine activity or ambulation, compared with control values.
Brain Morphometry Evaluation:
Cohort 2A
No test item-related effects were seen in the brain morphometry parameters.
The length and width of the brain, as measured grossly, were compared between controls and animals at 900 mg/kg/day. The mean values were similar and were considered not to show a test item-relationship.
Cohort 2B
The only statistically significant difference was that the thickness of the cerebral cortex (motor) in males was ~7% less than controls at 900 mg/kg/day. This was considered due to chance and not test item-related, because other parameters were not affected and in females, the thickness of the cerebral cortex (motor) was not statistically significantly less at 900 mg/kg/day compared to the controls (it was only ~5% less).
Other brain morphometry parameters at 900 mg/kg/day were different from the control means, but were within 10% of the controls and not statistically significant, so were considered not test item-related.
The length and width of the brain, as measured grossly, were compared between controls and animals at 900 mg/kg/day. The mean values were similar and were considered not to show a test item-relationship. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 900 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- reduction in number of live offspring
- changes in sex ratio
- fetal/pup body weight changes
- changes in litter size and weights
- changes in postnatal survival
- external malformations
- skeletal malformations
- visceral malformations
- Key result
- Abnormalities:
- no effects observed
- Developmental effects observed:
- no
- Conclusions:
- The administration of Omnirad 184 once daily (or twice daily 3 hours apart on some occasions) via
oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no
test item-related deaths. There were occasional observations of decreased activity, abnormal, un
coordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose le
vel-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body we
ight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/
day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. Ther
e were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and
F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity
was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirme
d by overall ambulation values, or any associated clinical findings, detailed functional observations or
brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/
kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 male
s only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels
(up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values
or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target
organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertro
phy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney
weights were seen in both sexes. In neurological investigations, there were no test item-related brain
weight differences, gross or histological findings or effects on the brain dimensions. There were no
histological findings in the reproductive tissues of the females that had not been observed to litter tha
t would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1
adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the i
n-life and pathological findings were considered not to be adverse. For F1 and F2 developmental para
meters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on
the pup numbers, survival or development. - Executive summary:
The objective of this study was to determine the potential toxicity of the chemical Omnirad 184, when given by oral gavage to adult rats and their offspring. This study was designed to provide an evaluation of reproduction and development as well as a thorough evaluation of systemic toxicity in pregnant and lactating females and their offspring. Detailed examination of key developmental endpoints, such as offspring viability, neonatal health, developmental status at birth, physical and functional development until adulthood, nervous and immune system development was expected to identify any specific target organs in the offspring. In addition, the study provided information about the effects of Omnirad 184 on the integrity and performance of the adult male and female reproductive systems and a neurobehavioural assessment of the F1 generation.
The study design was as follows:
Text Table 1 Experimental Design – F0 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
25
25
2
Omnirad 184
100
10
10
10.03
100
25
25
3
Omnirad 184
300
10
30
30.09
301
25
25
4
Omnirad 184
900
10
90
90.27
903
25
25
a Dose volume was based on the most recent body weight measurement.
Control/vehicle = 0.5% (w/v) Carboxymethylcellulose (CMC) medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 2 Experimental Design – F1 Unselected PND (Post Natal Day) 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 3 Experimental Design – Cohort 1A (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Omnirad 184
100
10
10
10.03
100
20
20
3
Omnirad 184
300
10
30
30.09
301
20
21d
4
Omnirad 184
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and
450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
d One female given 300 mg/kg/day died within minutes of receiving its first dose on Day 1, and another female
was added to the group.Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 4 Experimental Design – Surplus F1 Animals
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Omnirad 184
100
10
10
10.03
100
10
10
3
Omnirad 184
300
10
30
30.09
301
10
10
4
Omnirad 184
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 5 Experimental Design – Cohort 1B (Reproductive)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
20
20
2
Omnirad 184
100
10
10
10.03
100
20
20
3
Omnirad 184
300
10
30
30.09
301
20
20
4
Omnirad 184
900bc
10bc
90bc
90.27
903
20
20
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 6 Experimental Design – F2 Unselected PND 21 Pups
Group No.a
Number of Animalsb
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Unselected pups were not dosed, group number refers to dam group number.
b These pups had TSH and T4 blood sampling and necropsy procedures.
Text Table 7 Experimental Design – Cohort 2A (Neurobehavioural)
Group No.
Treatment
Dose Level (mg/kg/day)
Dose Volumea
(mL/kg)
Dose Conc.
(mg/mL)
Corrected Dose Conc.
(mg/mL)
Corrected Dose Levels (mg/kg/day)
Number of Animals
Males
Females
1
Control
0
10
0
0
0
10
10
2
Omnirad 184
100
10
10
10.03
100
10
10
3
Omnirad 184
300
10
30
30.09
301
10
10
4
Omnirad 184
900bc
10bc
90bc
90.27
903
10
10
a Dose was based on the most recent body weight measurement.
b From 19 Aug 2021, F1 Group 4 animals were dosed twice daily (approximately 3 hours apart) at 10 mL/kg and 450 mg/kg (45 mg/mL), equivalent to 900 mg/kg/day.
c From 30 Aug 2021, F1 Group 4 animals were dosed once daily at 10 mL/kg and 900 mg/kg/day.
Control/vehicle = 0.5% (w/v) CMC medium viscosity and 0.1% (w/v) Tween 80 in Milli-Q water
Conc. = concentrationText Table 8 Experimental Design – Cohort 2B (Neuropathology)a
Group No.
Number of Animals
Males
Females
1
10
10
2
10
10
3
10
10
4
10
10
a Cohort 2B were not dosed as they were terminated on PND 21/22.
In Week 17 of the study (F0 dosing and the start of F1 dosing), the 900 mg/kg/day dosing formulation could not be pulled up into the gavage tube being used for newly weaned pups, due to an unknown viscosity property, resulting in some subsets of F0 and/or F1 animals either not being dosed or being given the 300 mg/kg/day formulation either once or twice daily instead (refer to section 4.6.1 for details). From 19 Aug 2021, Group 4 F1 animals were dosed twice daily 3 hours apart at 450 mg/kg (equivalent to 900 mg/kg/day) until they were old enough for the next size gavage tube on 30 Aug 2021. Although there were occasions when animals were not dosed or received a dose level lower than that required by the study protocol, there was considered to be no overall impact on the results or study outcome due to the length of time that each generation was administered the test item dose levels correctly.
The following parameters and end points were evaluated in this study: clinical observations, body weights, food consumption, estrous cycles, mating performance, fertility indices, duration of gestation and overall litter performance, litter survival indices, litter and pup weights, pre-weaning physical development of F1 pups, assessments of sexual maturation of F1 animals, clinical pathology parameters (haematology, coagulation, clinical chemistry, and urinalysis), thyroid stimulating hormone (TSH) and thyroxine (T4) analysis, gross necropsy findings, splenic immunophenotyping analysis, organ weights, sperm evaluation, ovarian follicle counts and histopathological examinations.
None of the unscheduled deaths in any generation were considered to be test item-related.
For the F0 generation at 900 mg/kg/day, test item-related clinical observations of decreased activity, abnormal/uncoordinated gait and low carriage were occasionally seen for males and/or females before pairing, and decreased activity only during lactation. For F0 and F1 generations, salivation and/or ploughing were noted for all animals given ≥300 mg/kg/day and for several animals given 100 mg/kg/day, throughout the dosing periods. F0 and F1 males given 900 mg/kg/day had slightly lower body weight gain, and F0 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day.
There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development.
For males at 900 mg/kg/day, during the last 20 minutes of the 1 hour session, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively. Ambulation values were also lower for the same time period, but this was not confirmed by the overall ambulation values. As there were no clinical findings or detailed functional observations recorded that could have been associated with this, the cause of this apparent reduction in motor activity could not be determined.
For F0 males given ≥300 mg/kg/day and F0 females given ≥100 mg/kg/day, triglycerides were lower by up to 0.75-fold, and for males only, cholesterol higher by up to 1.3-fold. For F1 animals at ≥100 mg/kg/day, triglycerides were lower than controls by up to 0.62-fold, and at 900 mg/kg/day only, cholesterol higher by up to 1.25-fold. There were slightly higher TSH levels (up to 3.1-fold of controls) for samples taken on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day.
In the F0 animals dosed from 10 weeks prior to mating and continuing throughout mating and gestation until the females were at least LD 21, microscopically, centrilobular hypertrophy in the liver was considered test item-related in all dose groups in males but only at 900 mg/kg/day in females and mucosal hyperplasia of the glandular region, appearing as an increased number of goblet cells, was considered test item-related at ≥ 300 mg/kg/day in males and only at 900 mg/kg/day in females. Higher liver and kidney weights were seen in both sexes at all dose levels.
In the F1 cohort 1A animals, dosed from PND 21 until at least PND 90, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. In the stomach, there was mucosal hyperplasia of the glandular regions, appearing as an increased number of goblet cells. This finding was considered test item-related in males and females at ≥ 300 mg/kg/day. Liver weights were higher at ≥ 300 mg/kg/day in males and females. In addition, higher kidney weights were seen in females at all dose levels and at ≥ 300 mg/kg/day in males.
In the F1 cohort 1B animals, that were used to produce an F2 generation, and that were dosed from PND 21 until at least LD 21, microscopically there was diffuse centrilobular hypertrophy in the liver. This finding was considered test item-related at all dose levels in males and only at ≥ 300 mg/kg/day in females. Liver weights were higher at all dose levels in males and only at 900 mg/kg/day in females. In addition, higher kidney weights were seen in animals dosed at 900 mg/kg/day. Stomachs were not assessed microscopically in these animals. There were no test item-related organ weight differences in the female reproductive tissues.
There were no test item-related organ weight differences in a limited list of tissues in F1 and F2 pups that were euthanised at PND 21 without active dosing.
In the F1 cohort 2A and 2B animals used for neurological investigations (euthanised on PND 21 or dosed from PND 21 until at least PND 78-80), there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions.
There were no test item-related gross findings in the entire study, and there were only 6 premature deaths, all of which were considered not test item-related.
There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility.
In conclusion, administration of Omnirad 184 by once daily (or twice daily 3 hours apart on some occasions) oral gavage was well tolerated in Han Wistar rats at dose levels of up to 900 mg/kg/day with no test item-related deaths. There were occasional observations of decreased activity, abnormal, uncoordinated gait and low carriage for F0 animals at 900 mg/kg/day and for F0 and F1 animals, dose level-related salivation and/or ploughing. F0 and F1 males at 900 mg/kg/day had slightly lower body weight gain, and F0 and F1 females a slightly higher dose level-related body weight gain at ≥ 100 mg/kg/day. Food consumption was generally slightly higher for males and females at ≥ 100 mg/kg/day. There were no test item-related effects on F0 and F1 generation fertility, mating or reproduction, or F1 and F2 generation pup numbers, survival or development. For males at 900 mg/kg/day, motor activity was 9.6% or 12.7% lower overall for basic or fine movements, respectively, but this was not confirmed by overall ambulation values, or any associated clinical findings, detailed functional observations or brain morphometry values for Cohort 2A. Triglyceride values were lower for F0 males given ≥300 mg/kg/day, F0 females and F1 animals given ≥100 mg/kg/day, and cholesterol values higher for F0 males only at ≥300 mg/kg/day and F1 animals at 900 mg/kg/day. There were slightly higher TSH levels (up to 3.1-fold of controls) on the morning of necropsy in F0 and F1 adult males given 300 or 900 mg/kg/day. There were no test item-related effects on haematology, coagulation, urinalysis, T4 values or the immune cell populations analysed for immunophenotyping, at up to 900 mg/kg/day. Target organ effects were observed at dose levels of ≥100 mg/kg/day and consisted of centrilobular hypertrophy in the liver and stomach mucosal hyperplasia of the glandular region. Higher liver and/or kidney weights were seen in both sexes. In neurological investigations, there were no test item-related brain weight differences, gross or histological findings or effects on the brain dimensions. There were no histological findings in the reproductive tissues of the females that had not been observed to litter that would have caused reduced fertility. The NOAEL (No Observed Adverse Effect Level) for F0 and F1 adult toxicity, male and female fertility and reproduction was considered to be 900 mg/kg/day as the in-life and pathological findings were considered not to be adverse. For F1 and F2 developmental parameters, the NOAEL was considered to be 900 mg/kg/day as there were no test item-related effects on the pup numbers, survival or development.
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2014
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Details on test animals or test system and environmental conditions:
- - Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: 10-12 weeks
- The body weight of the pregnant animals on gestation day 0 varied between 148.5 – 191.5 g
- Housing: singly
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: The animals were acclimated to the laboratory conditions between start of the study (beginning of the experimental phase) and first administration (GD 6).
- Fasting: 16h before administration
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- oral: gavage
- Vehicle:
- CMC (carboxymethyl cellulose)
- Remarks:
- 1%
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
The aqueous test substance preparations were prepared at the beginning of the administration period and thereafter at maximum intervals of 7 days, which took into account the period of established stability. The preparation was stored in a freezer. - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Analytical verifications of the stability of the test substance in 1% Carboxymethylcellulose in drinking water over a period of a maximum of 7 days at room temperature were carried out prior to the start of the study. Samples of the test substance preparations were sent to the analytical laboratory at the beginning of administration for verification of the concentrations. The samples were also used to verify the homogeneity of the low and the high concentrations.
- Details on mating procedure:
- - Impregnation procedure: purchased timed pregnant
- Duration of treatment / exposure:
- gestation days 6 to 19
- Frequency of treatment:
- daily
- Dose / conc.:
- 50 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 150 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 500 mg/kg bw/day (actual dose received)
- No. of animals per sex per dose:
- 25
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: range finding study in pregnant rats
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: wice a day on working days or once a day on Saturdays, Sundays or on public holidays (GD 0-20).
DETAILED CLINICAL OBSERVATIONS: Yes
A cage-side examination was conducted at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity. If such signs occurred, the animals were examined several times daily (GD 0-20).
BODY WEIGHT: Yes
- Time schedule for examinations: GD 0, 1, 3, 6, 8, 10, 13, 15, 17, 19 and 20.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
The consumption of food was recorded for the intervals GD 0-1, 1-3, 3-6, 6-8, 8-10, 10-13, 13-15, 15-17, 17-19 and 19-20.
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uteri and the ovaries
OTHER: - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: live fetuses, dead fetuses - Fetal examinations:
- - External examinations: Yes: all per litter
- Soft tissue examinations: Yes: all per litter
- Skeletal examinations: Yes: all per litter
- Head examinations: Yes: all per litter - Statistics:
- Dunnett Test, Fishers exact test, Wilcoxon test
- Indices:
- conception rate (in %), preimplantation loss (in %), postimplantation loss (in %)
- Historical control data:
- 18 studies, from 2011 - 2013
- Details on maternal toxic effects:
- Maternal toxic effects:no effects
Details on maternal toxic effects:
Clinical examinations of the dams: Only pregnant dams were used for the calculations of mean maternal food consumption, body weight and body weight change. Only pregnant dams with scheduled sacrifice (GD 20) were used for the calculation of mean gravid uterine weights, corrected (net) body weight gain and summary of reproduction data. The following female was excluded from the above-mentioned calculations: Test group 3 (500 mg/kg bw/d): female No. 94 – not pregnant
Mortality: There were no test substance-related or spontaneous mortalities in any females of all test groups (0, 50, 150 or 500 mg/kg bw/d).
Clinical symptoms: Most females (20 out of 25) of the high-dose group (500 mg/kg bw/d) showed transient salivation during major parts of the treatment period. Salivation persisted in the respective animals for a few minutes after daily gavage dosing (i.e. about 10-15 minutes) and was initially observed on GD 9. Furthermore, high-dose female No. 95 (500 mg/kg bw/d) had an unsteady gait after treatment on GD 16. This finding persisted for approximately 2 hours. No clinical signs or changes of general behavior, which may be attributed to the test substance, were detected in any female at dose levels of 50 or 150 mg/kg bw/d during the entire study period.
Food consumption: The mean food consumption of the dams in test groups 1-3 (50, 150 or 500 mg/kg bw/d) was generally comparable to the concurrent control throughout the entire study period. This includes the statistically significantly increased food consumption value in the high-dose group on GD 15-19.
Body weight data: The mean body weights and the average body weight gains of the low-, mid- and high-dose rats (50, 150 or 500 mg/kg bw/d) were in general comparable to the controls throughout the entire study period. The statistically significantly increased body weight gain value in test group 2 on GD 0-6 (before start of treatment) is considered to be an incidental finding.
Corrected (net) body weight gain: The corrected body weight gain of test groups 1-3 (50, 150 and 500 mg/kg bw/d) revealed no difference of any biological relevance to the corresponding control group. Moreover, mean carcass weights remained also unaffected by the treatment.
Uterus weight The mean gravid uterus weights of the animals of test group 1-3 (50, 150 and 500 mg/kg bw/d) were not influenced by the test substance. The differences between these groups and the control group revealed no dose-dependency and were assessed to be without biological relevance.
Necropsy findings: No necropsy findings which could be attributed to the test substance were seen in any dam. There occurred two spontaneous findings in test group 1, i.e. one diaphragmatic hernia and one dilated renal pelvis. These findings were detected in single animals and therefore were not assessed to be treatment-related.
Reproduction data: The conception rate varied between 96% in test group 3 (500 mg/kg bw/d) and 100% in test groups 0-2 (0, 50 and 150 mg/kg bw/d). With these rates, a sufficient number of pregnant females were available for the purpose of the study (according to the test guidelines listed in chapter 2.3.). There were no test substance-related and/or biologically relevant differences between test groups 0, 1, 2 and 3 (0, 50, 150 and 500 mg/kg bw/d) in conception rate, in the mean number of corpora lutea and implantation sites or in the value calculated for the postimplantation loss, the number of resorptions and viable fetuses. However, the value calculated for the preimplantation loss was statistically significantly increased in the mid-dose group (150 mg/kg bw/d). Due to the lack of dose-response relationship and a historical control range which covers this value (HCD: 6.6% [3.2-15.8%]), this finding is considered as incidental and not related to treatment. All other observed differences are considered to reflect the normal range of fluctuations for animals of this strain and age. - Dose descriptor:
- NOAEL
- Effect level:
- 500 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: maternal toxicity
- Dose descriptor:
- NOAEL
- Effect level:
- 500 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: developmental toxicity
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
Sex distribution of the fetuses: The sex distribution of the fetuses in test groups 1-3 (50, 150 and 500 mg/kg bw/d) was comparable to the control fetuses. Any observable differences were without biological relevance.
Weight of the placentae: The mean placental weights in the high-dose group (500 mg/kg bw/d) were decreased (about 6-8% below control). Since the difference to the control was slight and as there were neither effects on intrauterine mortality nor on fetal weights, this apparent finding is not considered to be toxicologically relevant or adverse. The mean placental weights of the low- and mid-dose groups (50 and 150 mg/kg bw/d) were comparable to the corresponding control group.
Weight of the fetuses: The mean fetal weights of test groups 1, 2 and 3 (50, 150 and 500 mg/kg bw/d) were not influenced by the test substance and did not show any biologically relevant differences in comparison to the control group.
Fetal external malformations: No external malformations were recorded.
Fetal external variations: No external variations were recorded.
Fetal external unclassified observations: One unclassified external observation, i.e. blood coagulum around placenta, was recorded in two fetuses of the low-dose group (50 mg/kg bw/d). This finding was not considered biologically relevant.
Fetal soft tissue malformations: One soft tissue malformation was recorded for a control fetus. Male control fetus No. 13-08 had a situs inversus (abdominal and thoracic cavity). There were no soft tissue malformations in any of the treated groups.
Fetal soft tissue variations: Soft tissue variations were detected in all test groups including the control (0, 50, 150 or 500 mg/kg bw/d). The highest frequency was noted for dilated renal pelvis in all test groups including the control, and dilated ureter in the mid- and high-dose groups. The incidences of these findings were statistically significantly increased in the high-dose group. As a consequence, the total incidence of fetal soft tissue variations was also increased in this test group (Tab. 1.).
Fetal soft tissue unclassified observations: No unclassified soft tissue observations were recorded.
Fetal skeletal malformations: No skeletal malformations were recorded.
Fetal skeletal variations: For all test groups, skeletal variations of different bone structures were observed, with or without effects on corresponding cartilages. The observed skeletal variations were related to several parts of fetal skeleton and did not present a specific pattern. The overall incidences appeared without a relation to dosing and were comparable to the historical control data. For a better overview, all individual skeletal variations with statistically significant differences between the control and any treated group were compiled in the table below (Tab. 2). All incidences were expressed on a fetus per litter basis and any statistically significant differences, which were outside the historical control range were marked in bold types. As can be seen from Tab. 2. the rates of supernumerary thoracic vertebra and misshapen sternebra (unchanged cartilage) were statistically significantly increased in test group 3 (500 mg/kg bw/d) and slightly outside the historical control ranges. Concerning the other statistically significant findings, no dose dependency was observed and/or all values were clearly inside the historical control range, thus, an association to the test substance and a toxicological relevance is not assumed.
Fetal skeletal unclassified cartilage observations: Isolated cartilage findings without an impact on the respective bony structures, which were designated as unclassified cartilage observations, occurred in all test groups (Tab.3.). The observed unclassified cartilage findings were related to the skull, the sternum
and ribs and did not show any relation to dosing. However, the overall incidence of unclassified cartilage observations was statistically significantly increased in test group 3 (500 mg/kg bw/d), albeit only slightly above the historical control range (47.6 -80.3%).
Assessment of all fetal external, soft tissue and skeletal observations: External and skeletal malformations did not occur in any of the fetuses in this study. There was noted one soft tissue malformation in the control group, i.e. one situs inversus. External variations did not occur in any of the fetuses in this study. Some soft tissue variations and a range of skeletal variations were noted in all test groups including the controls. The majority of all variations were equally distributed about the different test groups, if normal biological variation is taken into account, and can be found in the historical control data at a comparable frequency. At the top dose (500 mg/kg bw/d) four different variations - dilated renal pelvis and dilated ureter as well as supernumerary thoracic vertebra and misshapen sternebra (unchanged cartilage) were statistically significantly increased and slightly above their historical control ranges. These rather minor increases resulted in a statistically significantly increased high-dose incidence if all different types of variations were summarized which just exceeded the historical control range (mean% affected fetuses per litter 50.44 - 56.61). Since these variations did not present a specific pattern and their rate was only slightly increased above an inherently high background rate, their toxicological significance is considered to be rather low. No unclassified soft tissue observations were recorded for any of the fetuses in this study. A spontaneous origin is assumed for the unclassified external observation and the unclassified skeletal cartilage observations which were observed in several fetuses of test groups 0, 1, 2 and 3 (0, 50, 150 and 500 mg/kg bw/d). The distribution and type of these findings do not suggest any relation to treatment. The total incidence of skeletal cartilage observations was slightly but statistically significantly higher in test group 3 (500 mg/kg bw/d) compared to control. However, since the control incidence was already at the upper limit of the historical control range (79.3 vs. 80.3 mean% affected fetuses per litter), this marginal increase is not considered to be biologically meaningful. Overall, fetal examinations revealed that there is no specific adverse effect of the test compound on fetal morphology at any of the tested dose levels. Particularly, there were no indications for a test substance-induced teratogenicity. - Remarks on result:
- not determinable due to absence of adverse toxic effects
- Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Executive summary:
In this guideline (OECD 414) study conducted with GLP certification, the test material (EC xxx-xxx-x) was determined to have a NOAEL of >= 500 mg/kg bw/day. The test substance doses (50-500 mg/kg bw/day) were administered via oral gavage to pregnant rats on the day of implantation until one day before the expected day of parturition. No differences of toxicological relevance between the control and the treated groups were observed. The results of the study indicate that the test material does not meet the criteria to be considered mutagenic under the EU Classification, Labelling, and Packaging (CLP) regulation (1272/2008).
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 08 Nov 2016 - 15 Dec 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Reason / purpose for cross-reference:
- reference to other study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Specific details on test material used for the study:
- Iracure 1173 (synonyms Darocur 1173, Omnirad 1173)
Chemical Identity: 2-hydroxy-2-methylpropiophenone
Batch number 0015591211
CAS number 7473-98-5
Expiry date 09 June 2019 - Species:
- rabbit
- Strain:
- New Zealand White
- Details on test animals or test system and environmental conditions:
- Sexually mature, virgin New Zealand White Rabblits (Crl:KBL(NZW)) were supplied at 16-17 weeks old. Only animals free from clinical signs of disease were used. The rabbits were housed singly in Type 4X03B700CP cages. The animals were accommodated in fully air-conditioned rooms at a maintained temeperature of 20+/-2C and a range of relative humidity of 30-70% The air exchange was 15 times per hour. The day/night cycle was generally 12 hours.
- Route of administration:
- oral: gavage
- Vehicle:
- CMC (carboxymethyl cellulose)
- Details on exposure:
- The test substance preparations were prepared at the beginning of the administration period and thereafter at intervals, which took into account the period of established stability. The preparations were kept at room temparature.
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Samples of the test substance preparations were sent to the analytical laboratory once during the study period (at the beginning of administration) for verification of the concentrations. These were analysed using GC. The samples taken for the concentration control analyses were also used to verify the homogeneity of the samples of the low and high concentrations each (15 and 150 mg/kg bw/d).
It was concluded that Irgacure was distrubuted homogeneously in 0.5% carboxymethyl cellulose in drinking water. The results of the GC analysis demonstrated the correctness of the concentrationa of Irgacure 1173 in 0.5% carboxymethyl cellulose in drinking water. - Details on mating procedure:
- After an acclimatization period of a least 5 days, the does were fertilzed by means of artifical insemination (AI). A synthetic hormone that stimulates release of LH and FSH from the anterior pituitary lobe was injected intramuscularly to the female rabbits about an hour before insemination. The ejaculate samples used for AI were obtained from male rabbits of the same breed as the females. The day of insemination was designated as GD 0 (beginning of the study) and the following day as GD1.
- Duration of treatment / exposure:
- The test substance was administered to the animals by oral gavage from implantation to one day prior to the expected day of parturition (GD 6-28) always at aprroximately the same time of day. The animals in the control group were treated in the same way with the vehicle (0.5% CMC) The volume administered eah day was 10 ml/kg body weight.
- Frequency of treatment:
- once a day
- Duration of test:
- 28 days
- Dose / conc.:
- 15 mg/kg bw/day (actual dose received)
- Remarks:
- low dose level
- Dose / conc.:
- 50 mg/kg bw/day (actual dose received)
- Remarks:
- mid-dose level
- Dose / conc.:
- 150 mg/kg bw/day (actual dose received)
- Remarks:
- high-dose level
- No. of animals per sex per dose:
- 25 female rabbits per dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- The test material was tested for its prenatal developmental toxicity in New Zealand White rabbits. The test material was administered as an aqueous suspension to groups of 25 inseminated female rabbits orally by gavage in doses of 15,50 and 150 mg/kg body weight/day on gestation days (GD) 6 to 28. The vehicle control group, consisting of 25 females, was dosed with the vehicle 0.5% carboxymethylcellulose suspension in drinking water (0.5% CMC) in parallel. A standard dose volume of 10mL/kg body weight was used for each group. At terminal sacrifice on G 29, 22-25 females per group had implantation sites.
- Maternal examinations:
- A cage-side examination was conducted at least once a day for signs of morbidity,pertinent behavioral changes and signs of overt toxicity. If such signs were observed the animals were examined several times daily (GD 0-29). During the administration period (GD 6-28) all animals were checked daily for any abnormal clinical signs before the administration and within 5 hours after administration.
- Ovaries and uterine content:
- The uteri and ovaries were removed and the following data were recorded: weight of the unopened uterus, number of the corpora lutea, number and the distribution of implantation sites classified as, live fetuses or dead implantations. The dead implanations were classed as either early resorptions, late resorptions or dead fetuses.
- Fetal examinations:
- At necropsy each fetus was weighed and examined macroscopically for external findings. The viability of the fetuses and the condition of the placentas, umbilical cords, fetal membranes, and fluids were examined. Individual placental weights were recorded. After the fetuses had been sacrificed, the abdomen and thorax were opened in order to examine the organs in situ before they were removed. The heart and the kidneys were sectioned in order to evaluate the internal structure. The sex of the fetuses was determined by examination of the gonads in situ.
Transverse sections were prepared of the heads.of approximately half the fetuses per doe. Transversal incision into the frontal/parietal head bones was made of the intact fetues, After fixation in ethyl alcohol, the skeletons were stained and investigated individually. - Statistics:
- Please see attachments titles Statistics
- Historical control data:
- Available for the following end points data collected between 1 Jan 2012 and 31 May 2017
Please see attachment entitled Historical control data
Mean Maternal Body weights during gestation (grams)
Reproduction data
Placenta weights (grams)
Mean fetal weights (grams)
Fetal external malformations
Fetal external variations
Fetal external unclassified findings
Fetal soft tissue malformations
Fetal soft tissue variations
Fetal soft tissue unclassified findings
Fetal skeletal variations
Fetal skeletal malformations
Fetal Skeletal cartilage - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Test group 3 - (150 mg/kg bw/d) unsteady gait and reduced attention shortly after treatment in all 25 females, splayed limbs after treatment in two females: all findings wore off between 2 and 5 hours post-dosing. Splayed limbs were recorded for 2 high-dose females shortly after treatment. These findings were considered as treatment related and adverse. However they disappeared 2-5 hours after dosing and were not observed before dosing the following day. Two females in mid range group had blood in bedding before and after treatment and subsequently died with in 48 hours.
Reduced defecation was observed in 4 control, 1 loe-dose, 4 mid-dose and two high dose females - displaying no dose related responce. There were no other clinical findings in the study. - Description (incidence and severity):
- NA
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- Test group 0 (0 mg/kg bw/d) - two animals died after gavage error
Test group 1 (15mg/kg bw/d)- one animal died after gavage error
Test group 2 (50 mg/kg bw/d) one aniimal died intercurrently
Test group 3 (150mg/kg bw/d) one animal died after gavage error - Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The mean body weights and the average body weight gain of the low-, mid- and high-dose groups (15, 50 and 150 mg/kg bw/d) were generally comparable to the concurrent control group throughout the entire study period. However, there were statistically significantly increased body weight gain values in the low- and the high-dose groups during GD 25-28 which were considered unrelated to treatment.
See illustration as attached - Mean body weight of pregnant animals
Also Table 3 - Mean Maternal body weight change during gestation (grams) - Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- The food consumption of the low-, mid- and high-dose rabbits (15, 50 and 150 mg/kg bw/d) was comparable to the concurrent control (0 mg/kg bw/d) throughout the entire study period.
- Food efficiency:
- no effects observed
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- effects observed, treatment-related
- Description (incidence and severity):
- Test group 3 - (150 mg/kg bw/d) unsteady gait and reduced attention shortly after treatment in all females, splayed limbs after treatment in two females: all finds wore off between 2 and 5 hours post-dosing.
Splayed limbs were recorded for 2 high-dose females shortly after treatment. - Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- The mean gravid uterus weight of the rabbits of the test groups was not influenced by the test substance.The differences between these groups and the control group showed no dose-dependency and were assessed to be without biological
relevance. Please also see table 4 - Mean Gravid weights and net maternal body weight change - Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Some spontaneous findings were noted in individual females of test groups 0, 2 and 3 (0, 50 and 150 mg/kg bw/d). These gross findings were:
• small spleen (additionally misshapen) in one control doe (No. 14 - 0 mg/kg bw/d).
• single erosion in stomach in one mid-dose doe (No. 56 - 50 mg/kg bw/d).
• increased reddish fluid in uterus in one high-dose doe (No. 93 - 150 mg/kg bw/d).
Findings in test groups 0, 1 and 3, such as thoracic cavity filled with either blood or serous fluid,
in two control does, one low-dose doe and one high-dose doe were indicative of gavage errors and were thus associated with unscheduled maternal deaths.
Please see table 5 - Summary of maternal necropsy observations
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- no effects observed
- Number of abortions:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test substance-related and/or biologically relevant differences between the different test groups in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the postimplantation losses, the number of resorptions and viable fetuses. All differences observed are considered to reflect the normal range of fluctuations for animals of this strain and age (see historical control dat as attached)
See Table 1 - summary of reproduction data
There were some foetues deaths found in test groups 0,1 and 3. None of these casualties were considered to be associated with treatment. - Pre- and post-implantation loss:
- no effects observed
- Description (incidence and severity):
- There were no test substance-related and/or biologically relevant differences between the different test groups in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the postimplantation losses, the number of resorptions and viable fetuses. All differences observed are considered to reflect the normal range of fluctuations for animals of this strain and age;
See Table 1 - summary of reproduction data - Total litter losses by resorption:
- no effects observed
- Description (incidence and severity):
- There were no test substance-related and/or biologically relevant differences between the different test groups in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the postimplantation losses, the number of resorptions and viable fetuses. All differences observed are considered to reflect the normal range of fluctuations for animals of this strain and age;
See Table 1 - summary of reproduction data - Early or late resorptions:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no test substance-related and/or biologically relevant differences between the different test groups in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the postimplantation losses, the number of resorptions and viable fetuses. All differences observed are considered to reflect the normal range of fluctuations for animals of this strain and age;
See Table 1 - summary of reproduction data - Dead fetuses:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Some dead fetuses were found in test groups 0, 1 and 3: one dead fetus in the litter of control, one in the litter of low-dose and two dead fetuses in the litter of high-dose. None of these casualties is considered to be associated with
treatment.
See Table 1 - summary of reproduction data
- Changes in pregnancy duration:
- no effects observed
- Description (incidence and severity):
- There were no observed changes in any group on the pregnancy duration
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): There were no observed effects in any group on the pregnancy duration - Changes in number of pregnant:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects on the number of females becoming pregnant. Conception rate in groups 1-4 were 88,92,100 and 100% respectively. Please see table 1 - Summary of reproduction data
- Other effects:
- no effects observed
- Description (incidence and severity):
- No other effects observed
- Details on maternal toxic effects:
- Clinical observations revealed toxicologically relevant effects at a dose of 150 mg/kg bw/d. All 25 high-dose females (150 mg/kg bw/d) showed unsteady gait and reduced attention at least once during the treatment period. Two high-dose females additionally displayed splayed limbs.
These clinical observations were made shortly after treatment and disappeared in the affected animals between 2 and 5 hours post-dosing. There were no persistent clinical findings. These clinical findings were considered as treatment-related and adverse.
The does at the lower dose levels (15 and 50 mg/kg bw/d) remained clinically unaffected by the test item.
Neither food consumption nor body weights/body weight gain (gross and corrected) of the does were affected by the treatment at any dose. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 50 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- clinical signs
- Abnormalities:
- no effects observed
- Fetal body weight changes:
- no effects observed
- Description (incidence and severity):
- The mean fetal weights of test groups 1, 2 and 3 were not influenced by the test substance and did not show any biologically relevant differences in comparison to the control group. Please see table 2 : mean placental and foeatal weights (on a litter basis)
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Please see table 2 : mean placental and foeatal weights (on a litter basis) - Reduction in number of live offspring:
- no effects observed
- Description (incidence and severity):
- Please see table 1 - Summary of reproduction data
- Changes in sex ratio:
- no effects observed
- Description (incidence and severity):
- The sex distribution of the fetuses in test groups 1-3 (15, 50 and 150 mg/kg bw/d) was comparable to the control fetuses. Any observable differences were without biological
relevance. Please see table 1 - Summary of reproduction data
- Changes in litter size and weights:
- no effects observed
- Description (incidence and severity):
- Please see table 2 : mean placental and foeatal weights (on a litter basis)
- Changes in postnatal survival:
- no effects observed
- External malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Three unclassified external observations were recorded in test groups 0, 1 and 3: hemorrhage in one fetus (150 mg/kg bw/d), placentae discolored in one dead fetus, each (0 mg/kg bw/d) and (15 mg/kg bw/d) and blood coagulum around placenta
in two dead fetuses (150 mg/kg bw/d). These findings are not considered to be related to treatment. Please see tables 6 and 7 Total Foetal external variation and total foetal external unclassified observations.
- Skeletal malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Skeletal malformations were detected in single fetuses of all test groups (0, 15, 50, 150 mg/kg bw/d), The incidence of these malformations was neither statisti-
cally significantly different from control nor dose-dependent and therefore, not considered biologically relevant. Please see table 11 - Total skeletal malformations.
For all test groups, skeletal variations of different bone structures were observed, with or without effects on corresponding cartilages. The observed skeletal variations were related to
several parts of fetal skeletons and appeared without a relation to dosing. The overall incidences of skeletal variations were comparable to the historical control data Please see table 12 and historical control data
For a better overview all skeletal variations with statistically significant differences between the control and the treated groups were complied in a table - please see table 13-statistically significant increses were not dose related and within historical control range.
- Visceral malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no external malformations in any foetus in the study. Soft tissue malformations occurred in all test groups including control (0, 15, 50 or 150 mg/kg bw/d). one Female low-dose fetus had multiple visceral malformations, i.e. high-arched
aortic arch, dilated aortic arch, narrowed pulmonary trunk and unexpanded lun, while one low dose foetus had a malpositioned kidney and a short ureter.Furthermore, female high-dose fetus showed microphthalmia on both eyes and severely fused sternebrae (bony plate). No ontogenetic pattern is recognizable for the individual malformations nor was there any cluster of any of these malformations seen in the other offspring of these test groups.
Other malformations, such as misshapen brain, small thymus, absent subclavian, cardiomegaly, absent gallbladder, fused cervical arch, thoracic hemivertebra and malpositioned and bipartite sternebra were scattered observations in individual fetuses of all test groups. They were not dose-related and some of them can be found in the historical control data. An associationof these findings to the treatment is not assumed.
Please see table 15 - total foetal malformations
The distribution of the findings about the test groups does not indicate an association to the treatment and no statistically significant differences between the groups were noted. The total
incidence of soft tissue malformations in treated animals did not differ significantly from the control group and was comparable to the historical control data. - Other effects:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The examination of the soft tissues revealed malpositioned carotid branches in individual fetuses of all test groups including the control group (0, 15, 50 or 150 mg/kg bw/d). An absent
lung lobe (Lobus inferior medialis) was noted in all substance-treated groups and showed a slightly but statistically significantly increased incidence in the low- and mid-dose groups (15
and 50 mg/kg bw/d). However, these are quite common findings in this rabbit strain, they can be found in the historical control data of the test facility at comparable incidences.
Other variations, such as cystic dilatation of the brain, dilated cerebral ventricle, malpositioned
subclavian artery origin, dilated aortic arch, blood-filled pericardium, small atrial chamber
(heart), enlarged atrial chamber (heart) and supernumerary gallbladder occurred in individual
offspring of all test groups including control (0, 15, 50 or 150 mg/kg bw/d). The incidences of
these variations were neither statistically significantly different from control nor dose-dependent
and, therefore, not considered biologically relevant . Some of them can be
found in the historical control data at comparable incidences
One unclassified soft tissue observation was recorded in one fetus of the high-dose group (150
mg/kg bw/d): blood coagulum around urinary bladder. This finding is not considered to be treat-
ment-related.
- Details on embryotoxic / teratogenic effects:
- No differences of toxicological relevance between the control and the treated groups (15, 50 and 150 mg/kg bw/d) were determined for any reproductive parameters, such as conception
rate, mean number of corpora lutea, mean number of implantations, as well as pre- and postimplantation loss and live litter size. Similarly, no adverse effect of the test substance on
uterine and placental weights as well as fetal weight and sex distribution of the fetuses was noted at any dose.
Overall, there were no toxicologically relevant adverse effects of the test substance on fetal
morphology at any dose.
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 150 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: here was no evidence for toxicologically relevant adverse effects of the test substance on fetal morphology at any dose, the no observed adverse effect level (NOAEL) for prenatal developmental toxicity is the highest dose of 150 mg/kg bw/d.
- Key result
- Abnormalities:
- no effects observed
- Key result
- Developmental effects observed:
- no
- Conclusions:
- Irgacure 1173 was administered to pregnant New Zealand White rabbits daily by stomach tube from implantation to one day prior to the expected day of parturition (GD 6-28) at dose levels of 15, 50 and 150 mg/kg bw/day.
Analyses confirmed the correctness of the prepared concentrations, as well as the homogeneous distribution and the stability of the test substance in the vehicle
Clinical observations revealed toxicologically relevant effects at a dose of 150 mg/kg bw/d. All 25 high-dose females (150 mg/kg bw/d) showed unsteady gait and reduced attention at least once during the treatment period. Two high-dose females additionally displayed splayed limbs. These clinical observations were made shortly after treatment and disappeared in the affected animals between 2 and 5 hours post-dosing. There were no persistent clinical findings. These clinical findings were considered as treatment-related and adverse. The does at the lower dose levels (15 and 50 mg/kg bw/d) remained clinically unaffected by the test item.
Neither food consumption nor body weights/body weight gain (gross and corrected) of the does were affected by the treatment at any dose. No differences of toxicological relevance between the control and the treated groups (15, 50 and 150 mg/kg bw/d) were determined for any reproductive parameters, such as conception rate, mean number of corpora lutea, mean number of implantations, aswell as pre- and postimplantation loss and live litter size. Similarly, no adverse effect of the test substance on uterine and placental weights as well as fetal weight and sex distribution of the fetuses was noted at any dose. Overall, there were no toxicologically relevant adverse effects of the test substance on fetal morphology at any dose.
Under the conditions of this prenatal developmental toxicity study, the oral administration of Irgacure 1173 to pregnant New Zealand White rabbits from implantation to one day prior to the expected day of parturition (GD 6-28) caused evidence of maternal toxicity at the high-dose (150 mg/kg bw/d), such as adverse clinical findings. In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 50 mg/kg bw/d.
Since there was no evidence for toxicologically relevant adverse effects of the test substance on fetal morphology at any dose, the no observed adverse effect level (NOAEL)for prenatal developmental toxicity is the highest dose of 150 mg/kg bw/d.
Under the conditions of this prenatal developmental toxicity study, the oral administration of Irgacure 1173 to pregnant New Zealand White rabbits from implantation to one day prior tothe expected day of parturition (GD 6-28) caused evidence of maternal toxicity at the high-dose (150 mg/kg bw/d), such as adverse clinical findings. In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 50 mg/kg bw/d. - Executive summary:
Irgacure 1173 was tested for its prenatal developmental toxicity in New Zealand White rabbits.The test substance was administered as an aqueous suspension to groups of 25 inseminated female New Zealand White rabbits orally by gavage in doses of 15, 50 and 150 mg/kg bodyweight/day (mg/kg bw/d) on gestation days (GD) 6 through 28. The vehicle control group, consisting of 25 females, was dosed with the vehicle 0.5% Carboxymethylcellulose suspension in drinking water (0.5% CMC) in parallel. A standard dose volume of 10 mL/kg body weight was used for each test group.
At terminal sacrifice on GD 29, 22-25 females per group had implantation sites.
Food consumption and body weight of the animals were recorded regularly throughout the study period. The state of health of the animals was checked each day.
On GD 29, all females were sacrificed and assessed by gross pathology (including weight determinations of the unopened uterus and placentas). For each doe, corpora lutea were counted and number and distribution of implantation sites (differentiated between resorptions,live and dead fetuses) were determined. The fetuses were removed from the uterus, sexed, weighed and further investigated for any external, soft tissue and skeletal (inclusive cartilage) findings.
RESULTS
Analytics
• The stability of the test substance in 1% Carboxymethylcellulose suspension in drinking water was demonstrated over a period of 7 days at room temperature.
• The homogeneous distribution of the test substance in the vehicle was shown.
• The correctness of the prepared concentrations was shown. Effects
The following test substance-related adverse effects/findings were noted:
Test group 3 (150 mg/kg bw/d):
Does
• Unsteady gait and reduced attention shortly after treatment in all females, splayed limbs after treatment in two females; all findings wore off between 2 and 5 hours post-dosing
Fetuses
• No test substance-related adverse effects on fetuses.
Test group 2 (50 mg/kg bw/d):
• No test substance-related adverse effects on does, gestational parameters or fetuses.
Test group 1 (15 mg/kg bw/d):
• No test substance-related adverse effects on does, gestational parameters or fetuses.
Under the conditions of this prenatal developmental toxicity study, the oral administration of Irgacure 1173 to pregnant New Zealand White rabbits from implantation to one day prior to the expected day of parturition (GD 6-28) caused evidence of maternal toxicity at the high-dose (150 mg/kg bw/d), such as adverse clinical findings. In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 50 mg/kg bw/d.
Since there was no evidence for toxicologically relevant adverse effects of the test substance on fetal morphology at any dose, the no observed adverse effect level (NOAEL) for prenatal developmental toxicity is the highest dose of 150 mg/kg bw/d.
Referenceopen allclose all
Table 1 - Summary of Body Weights (g)
F0 Males
| Sex: Male | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 124.9 | 176.6 | 225.1 | 268.9 | 303.1 | 333 | 355.5 |
| Group 1 | SD | 16.6 | 18.5 | 21 | 23.8 | 24.7 | 26.3 | 27.3 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 123.8 | 174.3 | 225.7 | 270.7 | 305.6 | 333.2 | 355.7 |
| Group 2 | SD | 16.3 | 18.8 | 23.4 | 25.2 | 26.4 | 28.5 | 30.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.9 | -1.3 | 0.3 | 0.7 | 0.8 | 0.1 | 0.1 | |
| 300 mg/kg/day | Mean | 121 | 171.9 | 222.1 | 268.3 | 302.8 | 332.4 | 355.9 |
| Group 3 | SD | 15.1 | 18.9 | 21 | 23.1 | 22.1 | 24.6 | 26.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -3.1 | -2.7 | -1.3 | -0.2 | -0.1 | -0.2 | 0.1 | |
| 900 mg/kg/day | Mean | 124.9 | 178.2 | 230.5 | 277.9 | 310.1 | 334.8 | 352.2 |
| Group 4 | SD | 11.6 | 14.2 | 18.9 | 21.9 | 24.1 | 25.7 | 28.3 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0 | 0.9 | 2.4 | 3.4 | 2.3 | 0.5 | -0.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 43 | 50 | 57 | 64 | 71 | 78 | 85 | ||
| 0 mg/kg/day | Mean | 374.2 | 389 | 406.9 | 417.8 | 427.7 | 428 | 435.6 |
| Group 1 | SD | 30.2 | 31.6 | 33.7 | 35 | 34.9 | 33.2 | 34.4 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 374.4 | 392.8 | 408 | 417.3 | 423.9 | 427.8 | 437.4 |
| Group 2 | SD | 32.8 | 34.9 | 36.1 | 35.5 | 36.7 | 37.4 | 37.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0.1 | 1 | 0.3 | -0.1 | -0.9 | -0.1 | 0.4 | |
| 300 mg/kg/day | Mean | 373.7 | 390.5 | 404.8 | 415.6 | 424 | 426.6 | 436.5 |
| Group 3 | SD | 29.8 | 32.2 | 33.2 | 33.8 | 36.9 | 36.4 | 38.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | 0.4 | -0.5 | -0.5 | -0.9 | -0.3 | 0.2 | |
| 900 mg/kg/day | Mean | 365.9 | 378.8 | 393.4 | 403.8 | 411.4 | 415.7 | 423.1 |
| Group 4 | SD | 29.7 | 32 | 33.1 | 34.7 | 35.5 | 34.6 | 36.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -2.2 | -2.6 | -3.3 | -3.3 | -3.8 | -2.9 | -2.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 92 | 99 | 106 | 113 | 120 | 127 | 134 | ||
| 0 mg/kg/day | Mean | 441.8 | 453.6 | 466.3 | 472.8 | 483.5 | 489.2 | 496.7 |
| Group 1 | SD | 35.9 | 36.1 | 37.8 | 37.8 | 38.9 | 39 | 41.7 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 441.6 | 450.2 | 462.2 | 470.4 | 478 | 487.7 | 492.6 |
| Group 2 | SD | 37.1 | 37.6 | 39.7 | 40.7 | 41.1 | 43.8 | 45.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | -0.7 | -0.9 | -0.5 | -1.1 | -0.3 | -0.8 | |
| 300 mg/kg/day | Mean | 446.7 | 452.6 | 463.6 | 475.6 | 483.9 | 492.7 | 496.6 |
| Group 3 | SD | 40.7 | 42.3 | 44.7 | 47 | 49.7 | 50.9 | 53.1 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 1.1 | -0.2 | -0.6 | 0.6 | 0.1 | 0.7 | 0 | |
| 900 mg/kg/day | Mean | 434.1 | 442.2 | 449.5 | 458.4 | 464.4 | 475.5 | 478.6 |
| Group 4 | SD | 38 | 39.8 | 40.3 | 40.7 | 40.7 | 42.5 | 44.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -1.7 | -2.5 | -3.6 | -3 | -3.9 | -2.8 | -3.6 | |
| Sex: Male | Day(s) Relative to Start Date | ||||
| 137 | 138 | 141 | 142 | ||
| 0 mg/kg/day | Mean | 507.9 | 501.8 | 493.1 | 473.4 |
| Group 1 | SD | 58.2 | 33.5 | 36.1 | 43.3 |
| N | 8 - | 6 - | 11 - | 5 - | |
| 100 mg/kg/day | Mean | 488 | 512.6 | 490.8 | 477.7 |
| Group 2 | SD | 46.5 | 39.6 | 50.5 | 63.9 |
| N | 5 | 7 | 13 | 7 | |
| %Diff | -3.9 | 2.1 | -0.5 | 0.9 | |
| 300 mg/kg/day | Mean | 458 | 534.4 | 491 | 494.8 |
| Group 3 | SD | 26.4 | 57.8 | 46.4 | 52.9 |
| N | 4 | 8 | 13 | 8 | |
| %Diff | -9.8 | 6.5 | -0.4 | 4.5 | |
| 900 mg/kg/day | Mean | 473.8 | 494.5 | 480.2 | 471.2 |
| Group 4 | SD | 41.8 | 27.5 | 54.1 | 65.1 |
| N | 8 | 4 | 13 | 5 | |
| %Diff | -6.7 | -1.5 | -2.6 | -0.5 | |
Table 2 - Summary of Body Weights (g)
F0 Females - Prior to mating
| Sex: Female | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 72.9 | 110.5 | 136.4 | 155.4 | 173.6 | 188.9 | 197.4 |
| Group 1 | SD | 13.5 | 13.9 | 14 | 13.8 | 13.3 | 13.9 | 14.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 76 | 114.9 | 140.8 | 162 | 181.1 | 195.7 | 206.3 |
| Group 2 | SD | 10.2 | 11.1 | 11.6 | 13.5 | 14.4 | 14.5 | 16.5 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 3.9 | 3.3 | 4.3 | 4.3 | 3.6 | 4.5 | |
| 300 mg/kg/day | Mean | 74.9 | 114.2 | 142.2 | 162.9 | 183.5 | 198.8 | 209.1* |
| Group 3 | SD | 12.4 | 11.1 | 10.6 | 12.3 | 12.8 | 14.7 | 15.6 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 2.8 | 3.4 | 4.3 | 4.9 | 5.7 | 5.2 | 5.9 | |
| 900 mg/kg/day | Mean | 77.2 | 116.4 | 147.2* | 171.3** | 191.6** | 208.2** | 214.9** |
| Group 4 | SD | 14.1 | 14.3 | 15.3 | 17.2 | 17.4 | 17.9 | 18.2 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 5.9 | 5.3 | 7.9 | 10.3 | 10.3 | 10.2 | 8.9 |
| Sex: Female | Day(s) Relative to Start Date | |||||
| 43 | 50 | 57 | 64 | 71 | ||
| 0 mg/kg/day | Mean | 207.3 | 214.6 | 222.2 | 226.2 | 230.5 |
| Group 1 | SD | 14.9 | 13.1 | 14.8 | 14.4 | 15.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 216 | 225.6 | 229.6 | 232.8 | 240.8 |
| Group 2 | SD | 17.1 | 18.4 | 19 | 20.5 | 20.5 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 5.2 | 3.3 | 2.9 | 4.5 | |
| 300 mg/kg/day | Mean | 217.2 | 227.9* | 234.7* | 238.4 | 245.9* |
| Group 3 | SD | 16.5 | 18.6 | 19.4 | 18.9 | 19 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.8 | 6.2 | 5.6 | 5.4 | 6.7 | |
| 900 mg/kg/day | Mean | 227.0** | 233.0** | 242.0** | 244.5** | 250.5** |
| Group 4 | SD | 17.2 | 17.5 | 18.6 | 19 | 19.7 |
| N | 24 | 24 | 24 | 24 | 24 | |
| %Diff | 9.5 | 8.6 | 8.9 | 8.1 | 8.7 |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 3 - Summary of Body Weights (g)
F0 Females - Gestation
| Sex: Female | Day(s) Relative to Mating (Litter: A) | ||||
| 0 | 7 | 14 | 20 | ||
| 0 mg/kg/day | Mean | 222.8 | 246.1 | 269.9 | 325.9 |
| Group 1 | SD | 14.5 | 13.5 | 16.3 | 24 |
| N | 18 - | 18 - | 18 - | 18 - | |
| 100 mg/kg/day | Mean | 237.2* | 259.5 | 282.3 | 338.5 |
| Group 2 | SD | 20.1 | 20.5 | 22.9 | 31.2 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 6.5 | 5.5 | 4.6 | 3.9 | |
| 300 mg/kg/day | Mean | 242.1** | 262.4* | 284.2 | 343.6 |
| Group 3 | SD | 20.5 | 22.4 | 24.6 | 28.1 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 8.6 | 6.6 | 5.3 | 5.4 | |
| 900 mg/kg/day | Mean | 243.7** | 264.5* | 291.1* | 352.5* |
| Group 4 | SD | 17.2 | 21.1 | 23.4 | 34.4 |
| N | 22 | 22 | 22 | 22 | |
| %Diff | 9.4 | 7.5 | 7.9 | 8.2 | |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 4 - Summary of Body Weights (g)
F0 Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | ||
| Sex | 7 | ||||||
| 1F | Mean | 261.7 | 274 | 275.1 | 282.8 | 280.1 | |
| SD | 19.1 | 20.8 | 19.3 | 21.2 | 16.6 | ||
| N | 23 | 22 | 23 | 23 | 23 | ||
| Mean | 268.2 | 280.9 | 283.3 | 290.9 | 288 | ||
| 2F | SD | 22.1 | 22.6 | 23 | 23.6 | 21.7 | |
| N | 23 | 24 | 24 | 24 | 24 | ||
| %Diff G1 | 2.5 | 2.5 | 3 | 2.9 | 2.8 | ||
| Mean | 270.8 | 282.7 | 285.6 | 294.8 | 291 | ||
| SD | 25.7 | 25 | 24.4 | 23.4 | 19.9 | ||
| 3F | N | 25 | 25 | 25 | 25 | 25 | |
| %Diff G1 | 3.5 | 3.2 | 3.8 | 4.2 | 3.9 | ||
| Mean | 271.6 | 286.4 | 292.2 | 302.6a | 300.1b | ||
| 4F | SD | 23.4 | 24 | 24.8 | 25.7 | 25.2 | |
| N | 22 | 22 | 22 | 22 | 22 | ||
| %Diff G1 | 3.8 | 4.5 | 6.2 | 7 | 7.1 | ||
Significantly different from control group 1 value :a=p≤0.05,b=p≤0.01 (Dunnett)
Table 5 - Summary of Body Weights (g)
F1 Animals - Cohort 1A
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
| 0 mg/kg/day | Mean | 56 | - | 71.8 | 119.9 | 172.1 | 219.5 | 269.6 |
| Group 1 | SD | 5.1 | - | 8.8 | 12.2 | 15.8 | 19.7 | 23.4 |
| N | 20 - | - | 20 - | 20 - | 20 - | 20 - | 20 - | |
| - | ||||||||
| 100 mg/kg/day | Mean | 56.3 | 61.0n | 70.8 | 116.4 | 166.9 | 213.7 | 261.2 |
| Group 2 | SD | 6.6 | 6.6 | 9 | 13.6 | 16.5 | 20.5 | 26.4 |
| N | 19 | 3 - | 18 | 20 | 20 | 20 | 20 | |
| %Diff | 0.7 | -1.4 | -2.9 | -3 | -2.6 | -3.1 | ||
| 300 mg/kg/day | Mean | 57.9 | 76.0n | 76.8 | 126.3 | 179.8 | 227 | 276.8 |
| Group 3 | SD | 4.9 | - | 9.3 | 14 | 16.9 | 19.6 | 24.9 |
| N | 20 | 1 - | 19 | 20 | 20 | 20 | 20 | |
| %Diff | 3.5 | 6.9 | 5.4 | 4.5 | 3.4 | 2.7 | ||
| 900 mg/kg/day | Mean | 54.7 | 59.0n 4.3 | 70.6 | 115.8 | 167.5 | 212.9 | 258.5 |
| Group 4 | SD | 5.5 | 6 - | 8.2 | 12.5 | 16.5 | 21.7 | 25.5 |
| N | 15 | 19 | 20 | 20 | 20 | 20 | ||
| %Diff | -2.2 | -1.6 | -3.4 | -2.7 | -3 | -4.1 |
[G] - Anova & Dunnett [I] - n - Inappropriate for statistics
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 41 | 48 | 55 | 62 | 68 | 69 | 70 | ||
| 0 mg/kg/day | Mean | 309.2 | 343.6 | 368.2 | 390.7 | 408.1 | 394 | 405.5 |
| Group 1 | SD | 26.7 | 31.5 | 33 | 37 | 40.1 | 42.2 | 40.7 |
| N | 20 - | 20 - | 20 - | 20 - | 14 - | 6 - | 20 - | |
| 100 mg/kg/day | Mean | 300.1 | 335.3 | 360.5 | 382.8 | 393 | 408.2 | 401.6 |
| Group 2 | SD | 29.4 | 32.6 | 35.7 | 36.5 | 35.3 | 42.7 | 34.3 |
| N | 20 | 20 | 20 | 20 | 16 | 6 | 16 | |
| %Diff | -2.9 | -2.4 | -2.1 | -2 | -3.7 | 3.6 | -1 | |
| 300 mg/kg/day | Mean | 317.3 | 349.4 | 376.1 | 396.8 | 411.1 | 445.9 | 411.1 |
| Group 3 | SD | 28 | 33 | 37.6 | 38.7 | 46.2 | 36.6 | 36.1 |
| N | 20 | 20 | 20 | 20 | 14 | 7 | 19 | |
| %Diff | 2.6 | 1.7 | 2.1 | 1.6 | 0.8 | 13.2 | 1.4 | |
| 900 mg/kg/day | Mean | 299.2 | 325.5 | 346.4 | 363.5 | 386.8 | 381.7 | 382.7 |
| Group 4 | SD | 29 | 32.1 | 37.5 | 40 | 40.6 | 48.2 | 38.9 |
| N | 20 | 20 | 20 | 20 | 14 | 11 | 15 | |
| %Diff | -3.2 | -5.3 | -5.9 | -7 | -5.2 | -3.1 | -5.6 |
Sex: Male |
| Day(s) Relative to Start Date |
71 | ||
0 mg/kg/day
Group 1 | Mean SD N
| - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 447.0n 69.3 2 - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | - - - - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | - - - - |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 4.9 20 - | - - - - | 66.0 7.8 20 - | 103.8 9.7 20 - | 136.5 10.7 20 - | 157.2 10.2 20 - | 176.5 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 55.3 4.9 19 5.3 | 58.0n 3.0 3 - | 68.9 5.7 18 4.5 | 106.1 9.4 20 2.2 | 138.3 11.9 20 1.3 | 159.7 14.1 20 1.6 | 178.8 16.8 20 1.3 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.0 5.0 21 4.7 | 71.0n - 1 - | 69.4 8.9 19 5.2 | 105.5 11.3 20 1.7 | 138.6 11.6 20 1.5 | 160.2 12.3 20 1.9 | 179.9 13.8 20 1.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 52.1 4.8 15 -0.8 | 56.3n 6.0 7 - | 66.1 8.2 18 0.2 | 103.2 10.6 20 -0.6 | 138.6 12.8 20 1.5 | 159.9 15.9 20 1.7 | 182.0 15.5 20 3.1 |
[G] - Anova & Dunnett
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 [G] | 48 [G] | 55 [G] | 62 [G] | 68 [G] | 69 [G] | 70 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.0 12.1 20 - | 203.2 14.4 20 - | 212.2 15.5 20 - | 221.0 18.8 20 - | 231.6 18.8 13 - | 228.1 18.9 7 - | 245.3n 11.0 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 192.4 18.5 20 1.8 | 205.7 17.3 20 1.2 | 214.1 18.0 20 0.9 | 224.2 20.5 20 1.4 | 235.2 22.6 17 1.6 | 216.7 6.8 3 -5.0 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 194.4 13.6 20 2.8 | 206.5 14.6 20 1.6 | 216.4 16.9 20 2.0 | 226.1 17.7 20 2.3 | 231.5 16.3 14 0.0 | 242.3 12.6 6 6.2 | 220.0n - 1 -10.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 201.6 17.1 20 6.7 | 209.2 17.6 20 3.0 | 218.0 17.7 20 2.7 | 226.5 18.3 20 2.5 | 237.5 19.1 15 2.6 | 235.6 26.0 5 3.3 | - - - - |
[G] - Anova & Dunnett
Sex: Female |
| Day(s) Relative to Start Date |
| ||
71 [G] | 72 [I] | 73 [G] | 74 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 226.9 16.6 8 - | - - - - | 228.6 21.1 5 - | 228.8 17.9 4 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 228.3 29.4 8 0.6 | 223.7n 12.2 3 - | 233.8 11.4 5 2.3 | 233.8 25.4 4 2.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 233.1 15.5 9 2.7 | 268.0n - 1 - | 225.0n 11.3 2 -1.6 | 237.7 20.5 7 3.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 235.0 23.9 10 3.6 | 247.0n 11.3 2 - | 232.8 19.2 6 1.9 | 235.0n 25.5 2 2.7 |
[G] - Anova & Dunnett: n - Inappropriate for statistics
Table 7 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Males
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 5.4 20 - | - - - - | 70.5 8.5 20 - | 116.5 12.0 20 - | 167.7 13.3 20 - | 212.9 17.3 20 - | 261.7 20.9 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 57.4 5.2 19 5.1 | 60.0n 8.5 3 - | 72.9 8.7 18 3.5 | 117.5 14.9 20 0.8 | 166.8 20.5 20 -0.6 | 211.2 25.9 20 -0.8 | 258.5 29.7 20 -1.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 56.8 5.7 20 4.0 | 70.0n - 1 - | 74.9 10.8 19 6.2 | 124.1 15.9 20 6.5 | 175.7 18.3 20 4.7 | 222.6 21.4 20 4.6 | 271.2 25.8 20 3.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 54.5 4.5 15 -0.1 | 59.3n 5.9 7 - | 69.9 8.7 18 -0.8 | 115.4 10.8 20 -0.9 | 166.8 12.3 20 -0.5 | 212.2 15.2 20 -0.3 | 258.6 18.7 20 -1.2 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 300.5 25.6 20 - | 334.6 30.5 20 - | 358.4 34.8 20 - | 381.5 38.4 20 - | 395.5 41.5 20 - | 410.3 44.0 20 - | 421.2 46.7 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 297.5 32.5 20 -1.0 | 330.9 34.9 20 -1.1 | 357.3 37.9 20 -0.3 | 379.6 39.8 20 -0.5 | 399.5 39.4 20 1.0 | 414.5 41.7 20 1.0 | 429.9 42.0 20 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 310.2 26.4 20 3.2 | 342.6 29.7 20 2.4 | 370.5 31.5 20 3.4 | 393.0 33.5 20 3.0 | 413.3 33.9 20 4.5 | 427.1 37.4 20 4.1 | 438.9 39.0 20 4.2 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 297.9 23.2 20 -0.8 | 324.7 26.1 20 -2.9 | 348.3 30.2 20 -2.8 | 365.3 33.5 20 -4.2 | 383.9 36.0 20 -2.9 | 399.6 35.5 20 -2.6 | 410.4 40.6 20 -2.6 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
90 | 97 | 104 | 111 | 118 | 125 | 128 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 431.7 46.1 20 - | 434.7 44.9 20 - | 441.9 43.5 20 - | 453.5 48.3 20 - | 464.3 49.9 20 - | 473.8 52.6 20 - | 484.1 58.9 8 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 440.9 44.6 20 2.1 | 444.3 45.0 20 2.2 | 452.5 46.6 20 2.4 | 463.5 46.1 20 2.2 | 477.7 47.1 20 2.9 | 492.3 48.2 20 3.9 | 494.2 58.5 11 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 448.1 41.2 20 3.8 | 452.0 38.8 20 4.0 | 456.2 39.7 20 3.2 | 471.1 40.7 20 3.9 | 483.8 43.3 20 4.2 | 494.5 45.3 20 4.4 | 515.1 42.9 9 6.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 415.5 40.8 20 -3.8 | 424.9 41.7 20 -2.3 | 428.4 41.3 20 -3.1 | 442.0 42.1 20 -2.5 | 451.7 42.8 20 -2.7 | 460.8 41.2 20 -2.8 | 463.5 45.3 12 -4.3 |
Sex: Male |
|
| Day(s) Relative to Start Date |
|
129 [G] | 130 [G] | 131 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 465.0 62.2 6 - | 475.3 45.0 9 - | - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 504.9 36.5 7 8.6 | 502.0 45.0 7 5.6 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 514.8 41.9 5 10.7 | 471.0 41.8 8 -0.9 | 485.5n 36.8 4 - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 476.3 43.3 8 2.4 | 436.3 10.1 3 -8.2 | 529.0n - 1 - |
Table 8 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Prior to Mating
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 5.1 20 - | - - - - | 66.2 7.1 20 - | 103.6 9.0 20 - | 136.1 9.5 20 - | 156.9 10.6 20 - | 174.8 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 53.8 7.9 19 2.4 | 57.3n 4.0 3 - | 66.6 10.4 18 0.7 | 103.4 13.9 20 -0.1 | 133.7 13.8 20 -1.7 | 155.3 14.0 20 -1.0 | 173.5 14.1 20 -0.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 4.4 20 6.0 | 68.0n - 1 - | 71.6 7.5 19 8.3 | 109.3 8.1 20 5.5 | 143.2 9.6 20 5.3 | 164.1 10.0 20 4.6 | 180.9 11.8 20 3.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 51.3 4.3 15 -2.3 | 56.3n 6.6 7 - | 64.6 7.4 18 -2.4 | 102.8 10.3 20 -0.7 | 136.6 9.8 20 0.4 | 157.6 13.1 20 0.4 | 176.8 13.7 20 1.1 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.6 11.3 20 - | 204.2 11.0 20 - | 210.9 11.8 20 - | 218.6 12.6 20 - | 227.6 11.8 20 - | 232.5 11.5 20 - | 236.2 12.5 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 188.1 15.8 20 -0.8 | 200.3 15.1 20 -1.9 | 209.8 15.4 20 -0.5 | 218.7 14.7 20 0.1 | 226.3 16.5 20 -0.6 | 230.7 18.2 20 -0.8 | 232.7 17.3 20 -1.5 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 199.2 12.4 20 5.1 | 211.3 13.2 20 3.5 | 220.1 14.6 20 4.4 | 227.0 15.2 20 3.9 | 236.9 15.5 20 4.1 | 243.1 16.4 20 4.5 | 246.7 17.5 20 4.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 195.6 14.1 20 3.2 | 203.7 14.7 20 -0.2 | 212.2 15.4 20 0.6 | 219.5 16.9 20 0.4 | 231.0 16.5 20 1.5 | 236.4 18.1 20 1.7 | 237.4 17.1 20 0.5 |
Sex: Female |
| Day(s) Relative to Start Date |
90 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 240.0 13.4 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 239.6 16.7 20 -0.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 251.2 17.1 20 4.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 242.7 18.8 20 1.1 |
Table 9 - Summary of Body Weights (g)
F1 Animals - Cohort 1B - Gestation
Sex: Female |
|
| Day(s) Relative to Mating (Litter: A) |
| |
0 | 7 | 14 | 20 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 241.9 12.7 19 - | 261.8 15.3 19 - | 284.1 15.0 19 - | 337.5 18.9 19 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 238.8 14.3 16 -1.3 | 258.2 14.3 16 -1.4 | 277.9 14.9 16 -2.2 | 334.6 19.9 16 -0.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 253.5 16.6 19 4.8 | 271.6 16.9 19 3.8 | 291.6 16.6 19 2.6 | 348.9 19.3 19 3.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 241.1 16.7 20 -0.3 | 260.8 20.7 20 -0.4 | 278.0 23.3 20 -2.1 | 336.5 32.1 20 -0.3 |
Table 10 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | |
| Sex | 7 | |||||
| 1F | Mean | 264.2 | 268.9 | 276.9 | 290.6 | 284.2 |
| SD | 14.4 | 12.9 | 14.9 | 16.8 | 17.9 | |
| N | 18 | 17 | 18 | 18 | 18 | |
| Mean | 262.3 | 271.3 | 275.8 | 288.6 | 286.3 | |
| 2F | SD | 14.5 | 15 | 14.2 | 16.6 | 15.4 |
| N | 16 | 15 | 16 | 16 | 16 | |
| %Diff G1 | -0.7 | 0.9 | -0.4 | -0.7 | 0.8 | |
| Mean | 278.4a | 288.5b | 292.6a | 306.2a | 302.1a | |
| SD | 18.9 | 16.6 | 15.7 | 15.6 | 16.2 | |
| 3F | N | 19 | 19 | 19 | 19 | 19 |
| %Diff G1 | 5.4 | 7.3 | 5.6 | 5.4 | 6.3 | |
| Mean | 265.9 | 276.3 | 283.6 | 292.4 | 291.5 | |
| 4F | SD | 21.6 | 24.5 | 20.3 | 21.7 | 22.9 |
| N | 20 | 19 | 20 | 20 | 19 | |
| %Diff G1 | 0.6 | 2.7 | 2.4 | 0.6 | 2.6 |
Table 11 - Summary of Body Weights (g)
F1 Animals - Cohort 2A
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 4.7 10 - | - - - - | 68.4 8.0 10 - | 115.1 11.9 10 - | 169.5 12.1 10 - | 219.4 13.6 10 - | 269.5 18.2 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.6 5.5 9 -0.1 | 59.0n 7.1 2 - | 67.0 8.9 9 -2.0 | 111.0 12.0 10 -3.6 | 160.7 12.0 10 -5.2 | 205.3 14.3 10 -6.4 | 250.8 18.2 10 -6.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 58.5 5.9 10 7.1 | 76.0n - 1 - | 74.7 10.9 9 9.2 | 122.9 14.3 10 6.8 | 177.1 16.3 10 4.5 | 224.4 19.6 10 2.3 | 273.9 23.0 10 1.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 55.4 3.6 9 1.5 | 59.0n 7.1 2 - | 67.3 9.2 9 -1.6 | 112.4 11.1 10 -2.3 | 164.3 12.4 10 -3.1 | 211.4 15.1 10 -3.6 | 259.5 21.4 10 -3.7 |
Sex: Male |
|
|
| Day(s) Relative to Start Date |
|
| |
41 | 48 | 55 | 58 | 59 | 60 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 308.0 21.9 10 - | 342.0 24.0 10 - | 368.4 25.7 10 - | 369.8 24.4 4 - | 396.8 32.0 5 - | 405.0 37.8 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 287.9 19.6 10 -6.5 | 320.4 24.2 10 -6.3 | 347.9 26.8 10 -5.6 | 376.8 42.4 4 1.9 | 369.8 20.2 6 -6.8 | 360.5 16.2 4 -11.0 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 313.5 26.9 10 1.8 | 348.1 27.9 10 1.8 | 372.8 25.9 10 1.2 | 403.3 32.3 6 9.1 | 404.0 54.1 3 1.8 | 369.5 14.5 4 -8.8 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 298.6 22.8 10 -3.1 | 326.5 27.2 10 -4.5 | 347.3 31.0 10 -5.7 | 362.0 46.0 4 -2.1 | 376.9 25.2 7 -5.0 | 341.5n 0.7 2 -15.7 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 51.5 4.2 10 - | - - - - | 62.9 6.5 10 - | 98.9 9.8 10 - | 128.6 9.3 10 - | 149.3 8.6 10 - | 166.7 9.1 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.2 2.9 9 5.3 | 54.0n 2.8 2 - | 65.4 5.5 9 4.0 | 100.5 8.6 10 1.6 | 131.3 8.2 10 2.1 | 155.1 8.6 10 3.9 | 172.1 10.8 10 3.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 7.4 10 8.2 | 76.0n - 1 - | 68.8 10.0 9 9.3 | 108.9 13.2 10 10.1 | 145.1** 12.9 10 12.8 | 167.6** 15.5 10 12.3 | 186.0** 18.3 10 11.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 53.6 3.7 9 4.0 | 57.0n 5.7 2 - | 63.2 5.3 9 0.5 | 100.5 6.2 10 1.6 | 135.7 10.3 10 5.5 | 159.2 10.4 10 6.6 | 179.3 12.7 10 7.6 |
Sex: Female |
|
| Day(s) Relative to Start Date |
|
| ||
41 [G] | 48 [G] | 55 [G] | 58 [G] | 59 [G1] | 60 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 179.8 10.1 10 - | 193.9 9.9 10 - | 203.7 10.7 10 - | 204.8 13.1 4 - | 214.0 12.7 5 - | 214.3 12.2 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 185.7 12.3 10 3.3 | 199.7 13.1 10 3.0 | 210.3 14.1 10 3.2 | 206.8 12.0 4 1.0 | 218.0 20.1 6 1.9 | 210.8 12.0 4 -1.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 202.3** 20.4 10 12.5 | 215.0** 20.3 10 10.9 | 223.8* 23.4 10 9.9 | 237.2 33.5 6 15.8 | 247.0 47.1 3 15.4 | 223.5 14.2 4 4.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 198.6* 14.7 10 10.5 | 210.7* 14.0 10 8.7 | 218.6 13.4 10 7.3 | 223.0 22.0 4 8.9 | 229.9 10.5 7 7.4 | 220.7 6.0 3 3.0 |
Tables 12. Summary of Clinical Chemistry Values
F0 Animals - Males. Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G1] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 71.4 12.3 10 - | 51.0 11.7 10 - | 128.5 12.4 10 - | 1.5 0.0 10 - | 217.1 68.4 10 - | 1.25 0.00 10 - | 7.35 0.66 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 74.6 10.3 9 1.04 | 49.1 12.0 9 0.96 | 126.0 25.8 9 0.98 | 1.5 0.0 9 1.00 | 292.7 163.0 9 1.35 | 1.25 0.00 9 1.00 | 6.90 0.68 9 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 60.5 * 4.9 10 0.85 | 44.5 7.8 10 0.87 | 125.1 32.5 10 0.97 | 1.5 0.0 10 1.00 | 213.2 70.8 10 0.98 | 1.25 0.00 10 1.00 | 7.16 0.35 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.6 * 5.4 10 0.86 | 36.8 14.1 9 0.72 | 115.4 28.5 10 0.90 | 1.5 0.0 10 1.00 | 196.8 60.0 10 0.91 | 1.25 0.00 10 1.00 | 6.94 0.68 10 0.94 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.2 3.2 10 - | 8.251 0.699 10 - | 1.789 0.249 10 - | 1.949 0.657 10 - | 66.29 1.89 10 - | 40.36 1.53 10 - | 25.93 2.18 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 32.1 3.5 9 0.94 | 8.778 1.195 9 1.06 | 1.853 0.393 9 1.04 | 1.933 0.681 9 0.99 | 66.09 3.31 9 1.00 | 41.22 2.83 9 1.02 | 24.87 2.61 9 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 * 2.9 10 0.87 | 8.113 0.521 10 0.98 | 2.170 * 0.404 10 1.21 | 1.594 0.199 10 0.82 | 66.67 2.46 10 1.01 | 41.93 1.91 10 1.04 | 24.74 2.11 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 31.5 3.6 10 0.92 | 8.208 0.634 10 0.99 | 2.327 ** 0.270 10 1.30 | 1.457 0.567 10 0.75 | 68.21 2.50 10 1.03 | 43.17 * 2.11 10 1.07 | 25.04 2.86 10 0.97 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.58 0.18 10 - | 2.618 0.059 10 - | 141.1 1.3 10 - | 4.98 0.29 10 - | 103.7 1.2 10 - | 1.404 0.193 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.69 0.22 9 1.07 | 2.637 0.076 9 1.01 | 141.4 1.9 9 1.00 | 4.90 0.34 9 0.98 | 103.7 1.4 9 1.00 | 1.531 0.178 9 1.09 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.71 0.17 10 1.08 | 2.654 0.055 10 1.01 | 140.3 2.2 10 0.99 | 4.97 0.15 10 1.00 | 102.3 1.8 10 0.99 | 1.545 0.152 10 1.10 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.75 0.28 10 1.11 | 2.716 ** 0.069 10 1.04 | 141.3 1.7 10 1.00 | 5.02 0.29 10 1.01 | 101.4 ** 2.0 10 0.98 | 1.591 0.187 10 1.13 |
Tables 13. Summary of Clinical Chemistry Values
F0 Animals - Females. Day: 115 Relative to Start Date
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 131.2 28.7 9 - | 117.1 17.5 9 - | 223.4 49.2 9 - | 1.5 0.0 9 - | 630.0 384.5 9 - | 1.25 0.00 9 - | 10.50 1.01 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 105.3 10.9 8 0.80 | 110.0 18.4 8 0.94 | 200.3 73.8 7 0.90 | 1.5 0.0 8 1.00 | 356.8 132.6 8 0.57 | 1.25 0.00 8 1.00 | 10.11 0.94 8 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 116.1 21.8 9 0.88 | 105.7 14.1 9 0.90 | 157.1 38.5 9 0.70 | 1.5 0.0 9 1.00 | 614.7 356.8 9 0.98 | 1.25 0.00 9 1.00 | 9.38 0.84 9 0.89 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 106.4 25.3 10 0.81 | 100.4 21.0 10 0.86 | 173.2 49.5 10 0.78 | 1.5 0.0 9 1.00 | 428.3 244.8 9 0.68 | 1.25 0.00 9 1.00 | 10.57 1.26 9 1.01 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G1] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 29.4 3.2 9 - | 4.740 1.150 9 - | 2.828 0.449 9 - | 0.589 0.061 9 - | 56.30 2.58 9 - | 40.21 1.79 9 - | 16.09 2.22 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 24.6 * 4.2 8 0.84 | 5.008 0.434 8 1.06 | 2.641 0.493 8 0.93 | 0.461 0.140 8 0.78 | 55.21 2.41 8 0.98 | 39.51 2.62 8 0.98 | 15.70 0.72 8 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 24.0 * 2.5 8 0.82 | 4.863 0.808 8 1.03 | 2.889 0.655 9 1.02 | 0.535 0.177 8 0.91 | 56.38 2.85 8 1.00 | 40.59 2.45 9 1.01 | 16.19 1.63 8 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 25.6 4.6 9 0.87 | 5.003 0.815 9 1.06 | 2.827 0.925 9 1.00 | 0.479 0.115 9 0.81 | 58.48 3.36 9 1.04 | 42.81 2.93 10 1.06 | 15.68 1.98 9 0.97 |
Sex: Female |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.53 0.34 9 - | 2.680 0.100 9 - | 140.7 2.1 9 - | 5.12 0.42 9 - | 99.8 2.2 9 - | 1.708 0.538 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.54 0.23 8 1.00 | 2.621 0.139 8 0.98 | 140.0 3.5 8 1.00 | 5.05 0.57 8 0.99 | 100.1 1.9 8 1.00 | 1.649 0.518 8 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.50 0.29 8 0.99 | 2.694 0.096 9 1.01 | 140.1 1.3 9 1.00 | 5.24 0.62 9 1.02 | 99.3 1.7 9 1.00 | 1.538 0.510 8 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.78 0.47 9 1.10 | 2.781 0.183 9 1.04 | 140.1 2.3 10 1.00 | 5.19 0.42 10 1.01 | 97.8 2.9 10 0.98 | 1.949 0.766 9 1.14 |
Tables 14. Summary of Clinical Chemistry Values
F1 Animals - Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 68.5 6.0 10 - | 57.6 11.4 10 - | 153.4 23.5 10 - | 1.5 0.0 9 - | 239.9 42.0 9 - | 1.25 0.00 9 - | 7.02 0.68 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 63.0 7.2 10 0.92 | 52.0 7.9 10 0.90 | 153.5 23.8 10 1.00 | 1.5 0.0 10 1.00 | 239.2 101.7 10 1.00 | 1.25 0.00 10 1.00 | 6.32 0.62 10 0.90 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 61.3 * 5.3 10 0.89 | 49.9 8.6 10 0.87 | 143.2 27.9 10 0.93 | 1.5 0.0 10 1.00 | 210.5 55.4 10 0.88 | 1.25 0.00 10 1.00 | 6.84 0.59 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.2 * 4.0 10 0.89 | 42.5 ** 13.0 10 0.74 | 141.5 23.9 10 0.92 | 1.5 0.0 10 1.00 | 174.9 37.4 10 0.73 | 1.25 0.00 10 1.00 | 7.24 0.85 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 30.8 4.0 9 - | 9.268 0.551 9 - | 1.723 0.237 9 - | 2.458 1.001 9 - | 66.48 1.47 9 - | 41.30 1.39 10 - | 25.16 1.24 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 28.1 3.8 10 0.91 | 8.529 ** 0.254 10 0.92 | 1.894 0.316 10 1.10 | 2.030 0.674 10 0.83 | 66.44 1.28 10 1.00 | 41.71 2.18 10 1.01 | 24.73 2.00 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 2.6 10 0.97 | 8.735 0.590 10 0.94 | 2.007 0.325 10 1.16 | 1.784 0.635 10 0.73 | 65.87 2.39 10 0.99 | 41.11 2.06 10 1.00 | 24.76 2.03 10 0.98 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.6 3.7 10 1.06 | 8.617 * 0.503 10 0.93 | 2.147 * 0.325 10 1.25 | 1.522 0.615 10 0.62 | 65.97 1.76 10 0.99 | 42.49 1.31 10 1.03 | 23.48 1.78 10 0.93 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G1] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.64 0.10 9 - | 2.592 0.033 9 - | 141.0 0.8 10 - | 5.09 0.30 10 - | 100.5 0.5 10 - | 1.747 0.191 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.70 0.19 10 1.03 | 2.620 0.064 10 1.01 | 142.0 1.2 10 1.01 | 5.12 0.30 10 1.01 | 101.7 * 0.8 10 1.01 | 2.034 ** 0.122 10 1.16 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.67 0.19 10 1.02 | 2.626 0.074 10 1.01 | 141.1 1.5 10 1.00 | 4.99 0.20 10 0.98 | 101.3 1.2 10 1.01 | 1.968 ** 0.179 10 1.13 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.83 0.17 10 1.11 | 2.657 0.064 10 1.02 | 141.6 2.0 10 1.00 | 4.95 0.18 10 0.97 | 99.9 1.6 10 0.99 | 2.065 ** 0.116 10 1.18 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 78.0 10.7 7 - | 55.9 14.4 7 - | 100.6 26.1 7 - | 1.5 0.0 7 - | 295.7 114.0 7 - | 1.25 0.00 7 - | 8.09 1.15 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 71.9 7.5 9 0.92 | 54.6 17.9 9 0.98 | 93.2 23.3 9 0.93 | 1.5 0.0 9 1.00 | 242.9 83.5 9 0.82 | 1.25 0.00 9 1.00 | 7.43 0.78 9 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 68.3 6.1 10 0.88 | 49.9 13.2 10 0.89 | 94.3 24.1 10 0.94 | 1.5 0.0 10 1.00 | 259.2 98.3 10 0.88 | 1.25 0.00 10 1.00 | 7.70 0.89 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 73.7 9.9 10 0.94 | 59.8 26.8 10 1.07 | 135.4 38.1 10 1.35 | 2.2 1.5 10 1.47 | 296.8 105.5 10 1.00 | 1.25 0.00 10 1.00 | 7.66 1.24 10 0.95 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.0 3.5 7 - | 8.577 0.866 7 - | 1.589 0.267 7 - | 1.421 0.304 7 - | 65.96 2.49 7 - | 45.56 2.60 7 - | 20.40 2.41 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 34.8 4.5 9 1.02 | 7.801 0.566 9 0.91 | 1.512 0.297 9 0.95 | 1.021 0.261 9 0.72 | 67.76 2.46 9 1.03 | 46.70 1.56 9 1.03 | 21.06 1.80 9 1.03 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 32.7 4.0 10 0.96 | 8.161 0.464 10 0.95 | 1.647 0.235 10 1.04 | 1.268 0.529 10 0.89 | 68.21 2.48 10 1.03 | 46.77 2.62 10 1.03 | 21.44 2.18 10 1.05 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.4 4.5 10 0.95 | 8.082 0.391 10 0.94 | 1.913 * 0.234 10 1.20 | 0.978 0.268 10 0.69 | 67.82 2.61 10 1.03 | 46.97 2.19 10 1.03 | 20.85 1.87 10 1.02 |
Sex: Female |
|
| Reporting Biochemistry |
|
| ||
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.27 0.35 7 - | 2.574 0.035 7 - | 140.3 1.1 7 - | 4.43 0.42 7 - | 103.6 1.4 7 - | 1.227 0.104 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.24 0.21 9 0.99 | 2.581 0.060 9 1.00 | 140.2 1.2 9 1.00 | 4.78 0.29 9 1.08 | 102.9 1.2 9 0.99 | 1.436 0.206 9 1.17 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.18 0.30 10 0.96 | 2.591 0.053 10 1.01 | 140.6 0.8 10 1.00 | 4.45 0.20 10 1.00 | 104.1 1.3 10 1.01 | 1.379 0.205 10 1.12 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.30 0.28 10 1.01 | 2.664 ** 0.039 10 1.03 | 140.4 1.1 10 1.00 | 4.86 * 0.27 10 1.10 | 102.2 0.9 10 0.99 | 1.312 0.263 10 1.07 |
Table 15. Summary of Coagulation Values
F0 Animals – Males, Day: 137 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.55 0.20 10 - | 13.51 1.35 10 - | 2.278 0.177 10 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.43 0.15 10 0.99 | 12.52 1.72 10 0.93 | 2.282 0.122 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.30 ** 0.14 10 0.98 | 12.35 1.14 10 0.91 | 2.286 0.185 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.26 ** 0.12 10 0.97 | 11.77 1.51 10 0.87 | 2.454 * 0.114 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 16. Summary of Coagulation Values
F0 Animals – Females, Day: 115 Relative to Start Date
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.19 0.21 9 - | 11.17 1.22 9 - | 1.713 0.292 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.48 0.34 10 1.03 | 12.17 1.55 10 1.09 | 1.612 0.157 10 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.27 0.32 9 1.01 | 11.82 1.49 9 1.06 | 1.743 0.316 9 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.39 0.73 10 1.02 | 11.16 2.64 10 1.00 | 1.682 0.425 10 0.98 |
[G] - Anova & Dunnett
Table 17. Summary of Coagulation Values
F1 Animals - Cohort 1A, Day: 70 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.50 0.14 9 - | 12.27 1.80 9 - | 2.580 0.172 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.40 0.11 10 0.99 | 12.79 2.10 10 1.04 | 2.516 0.141 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.34 * 0.14 10 0.98 | 12.09 2.30 10 0.99 | 2.655 0.219 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.33 * 0.05 9 0.98 | 11.51 2.25 9 0.94 | 2.707 0.250 9 1.05 |
[G] - Anova & Dunnett: * = p ≤ 0.05
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.31 0.26 9 - | 12.21 1.52 8 - | 2.116 0.251 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.17 0.17 9 0.99 | 12.33 1.57 9 1.01 | 2.128 0.172 9 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.15 0.14 10 0.98 | 11.65 0.99 10 0.95 | 2.186 0.194 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.02 ** 0.17 10 0.97 | 11.71 1.65 10 0.96 | 2.182 0.099 10 1.03 |
[G] - Anova & Dunnett: ** = p ≤ 0.01
Table 18. Summary of Hematology Values
F0 Animals – Males Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G1] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 4.651 0.478 10 - | 0.961 0.257 10 - | 3.480 0.289 10 - | 0.065 0.014 10 - | 0.116 0.046 10 - | 0.008 0.004 10 - | 0.019 0.006 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 4.340 0.805 10 0.93 | 0.769 0.202 10 0.80 | 3.390 0.784 10 0.97 | 0.062 0.018 10 0.95 | 0.096 0.024 10 0.83 | 0.006 0.005 10 0.75 | 0.016 0.008 10 0.84 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.469 0.783 10 0.96 | 0.687 * 0.169 10 0.71 | 3.605 0.697 10 1.04 | 0.057 0.021 10 0.88 | 0.089 0.023 10 0.77 | 0.010 0.000 10 1.25 | 0.019 0.007 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.192 0.960 10 1.12 | 0.854 0.199 10 0.89 | 4.126 0.908 10 1.19 | 0.079 0.028 10 1.22 | 0.092 0.040 10 0.79 | 0.009 0.006 10 1.13 | 0.027 0.013 10 1.42 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 8.813 0.449 10 - | 153.6 4.7 10 . | 0.4464 0.0164 10 - | 50.68 1.37 10 - | 17.43 0.57 10 - | 343.9 7.0 10 - | 11.88 0.31 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.630 0.303 10 0.98 | 150.7 6.5 10 0.98 | 0.4441 0.0155 10 0.99 | 51.52 2.26 10 1.02 | 17.46 1.03 10 1.00 | 338.9 7.5 10 0.99 | 12.00 0.48 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.572 0.317 10 0.97 | 149.6 5.2 10 0.97 | 0.4388 0.0129 10 0.98 | 51.25 1.89 10 1.01 | 17.47 0.71 10 1.00 | 341.1 6.6 10 0.99 | 12.01 0.52 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.573 0.220 10 0.97 | 147.4 5.4 10 0.96 | 0.4364 0.0139 10 0.98 | 50.92 1.39 10 1.00 | 17.17 0.55 10 0.99 | 337.5 4.5 10 0.98 | 12.16 0.48 10 1.02 |
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 781.6 146.2 10 - | 155.83 19.62 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 765.3 76.5 10 0.98 | 142.51 41.33 10 0.91 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 760.4 92.6 10 0.97 | 160.37 18.17 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 883.4 160.3 10 1.13 | 171.47 21.32 10 1.10 |
Table 19. Summary of Hematology Values
F0 Animals – Females Day: 115 Relative to Start Date
Sex: Female |
| Reporting Hematology | ||||||
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.168 1.072 8 - | 1.669 0.559 8 - | 3.265 0.790 8 - | 0.109 0.031 8 - | 0.095 0.038 8 - | 0.011 0.006 8 - | 0.023 0.009 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.529 0.603 10 1.07 | 1.737 0.408 10 1.04 | 3.564 0.620 10 1.09 | 0.113 0.025 10 1.04 | 0.077 0.039 10 0.81 | 0.012 0.008 10 1.07 | 0.025 0.010 10 1.11 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 5.866 1.252 10 1.14 | 1.903 0.424 10 1.14 | 3.694 0.894 10 1.13 | 0.143 0.060 10 1.31 | 0.091 0.038 10 0.96 | 0.013 0.007 10 1.16 | 0.026 0.013 10 1.16 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.570 0.802 10 1.08 | 1.762 0.315 10 1.06 | 3.555 0.672 10 1.09 | 0.144 0.051 10 1.32 | 0.073 0.013 10 0.77 | 0.008 0.004 10 0.71 | 0.028 0.011 10 1.24 |
Sex: Female |
| Reporting Hematology | ||||||
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.958 0.449 8 - | 157.3 3.4 8 . | 0.4679 0.0124 8 - | 58.88 2.37 8 - | 19.80 0.92 8 - | 336.4 5.2 8 - | 12.00 1.06 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.678 0.380 10 0.96 | 153.3 5.2 10 0.97 | 0.4515 0.0208 10 0.97 | 58.86 1.56 10 1.00 | 19.99 0.77 10 1.01 | 339.8 8.3 10 1.01 | 11.84 0.79 10 0.99 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.466 0.606 9 0.94 | 152.7 5.1 10 0.97 | 0.4468 0.0194 9 0.95 | 59.99 2.69 9 1.02 | 20.48 1.21 9 1.03 | 341.6 7.1 9 1.02 | 11.82 0.97 10 0.99 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.528 0.566 10 0.95 | 151.3 8.8 10 0.96 | 0.4428 0.0316 10 0.95 | 58.85 1.83 10 1.00 | 20.10 1.07 10 1.02 | 341.5 10.8 10 1.02 | 12.17 1.05 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 894.6 119.2 8 - | 177.99 20.44 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 794.3 132.8 10 0.89 | 178.18 31.61 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 872.4 121.5 9 0.98 | 161.09 22.77 10 0.91 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 833.4 177.2 10 0.93 | 173.25 30.61 10 0.97 |
Table 20. Summary of Hematology Values
F1 Animals – Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 6.989 1.038 10 - | 0.930 0.183 10 - | 5.762 0.940 10 - | 0.121 0.043 10 - | 0.108 0.036 10 - | 0.008 0.009 10 - | 0.059 0.018 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.203 0.765 10 1.03 | 0.970 0.322 10 1.04 | 5.976 0.743 10 1.04 | 0.099 0.031 10 0.82 | 0.091 0.031 10 0.84 | 0.011 0.006 10 1.38 | 0.054 0.023 10 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.256 0.780 10 1.04 | 0.993 0.193 10 1.07 | 6.050 0.666 10 1.05 | 0.086 0.030 10 0.71 | 0.074 0.023 10 0.69 | 0.007 0.005 10 0.88 | 0.047 0.009 10 0.80 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.090 * 0.978 10 1.16 | 1.081 0.253 10 1.16 | 6.736 1.071 10 1.17 | 0.115 0.030 10 0.95 | 0.080 0.032 10 0.74 | 0.011 0.006 10 1.38 | 0.064 0.023 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Hematology |
|
|
| |
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 9.078 0.461 10 - | 163.2 4.9 10 . | 0.4747 0.0207 10 - | 52.33 1.52 10 - | 17.99 0.90 10 - | 343.7 10.9 10 - | 11.47 0.49 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.551 * 0.488 10 0.94 | 156.9 * 5.4 10 0.96 | 0.4471 ** 0.0181 10 0.94 | 52.33 2.09 10 1.00 | 18.37 0.87 10 1.02 | 350.8 6.2 10 1.02 | 11.59 0.52 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.522 ** 0.298 10 0.94 | 157.1 * 4.1 10 0.96 | 0.4519 * 0.0171 10 0.95 | 53.06 1.54 10 1.01 | 18.44 0.54 10 1.03 | 347.7 7.0 10 1.01 | 11.63 0.51 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.340 ** 0.285 10 0.92 | 154.7 ** 4.4 10 0.95 | 0.4456 ** 0.0166 10 0.94 | 53.45 1.51 10 1.02 | 18.56 0.67 10 1.03 | 347.4 7.6 10 1.01 | 11.84 0.32 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 763.2 131.0 10 - | 193.85 15.23 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 876.8 89.5 10 1.15 | 171.56 23.85 10 0.89 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 899.7 76.1 10 1.18 | 198.49 23.89 10 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 823.3 144.5 10 1.08 | 208.78 30.71 10 1.08 |
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G1] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G1] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.104 1.013 9 - | 0.587 0.094 9 - | 4.321 0.997 9 - | 0.070 0.018 9 - | 0.081 0.019 9 - | 0.003 0.005 9 - | 0.038 0.010 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.207 1.111 9 1.02 | 0.671 0.142 9 1.14 | 4.371 1.058 9 1.01 | 0.053 0.017 9 0.76 | 0.074 0.028 9 0.92 | 0.004 0.005 9 1.33 | 0.029 0.014 9 0.76 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.656 0.870 10 0.91 | 0.708 0.238 10 1.21 | 3.780 0.791 10 0.87 | 0.060 0.021 10 0.86 | 0.067 0.020 10 0.83 | 0.003 0.005 10 0.90 | 0.034 0.008 10 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.515 1.387 10 1.08 | 0.681 0.323 10 1.16 | 4.640 1.417 10 1.07 | 0.076 0.039 10 1.09 | 0.079 0.031 10 0.97 | 0.004 0.005 10 1.20 | 0.037 0.014 10 0.98 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.842 0.436 9 - | 151.6 5.3 9 . | 0.4248 0.0164 9 - | 54.23 1.82 9 - | 19.38 0.95 9 - | 356.8 8.1 9 - | 10.70 0.38 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.868 0.225 9 1.00 | 148.8 6.5 9 0.98 | 0.4244 0.0173 9 1.00 | 53.91 1.31 9 0.99 | 18.89 0.64 9 0.97 | 350.7 8.5 9 0.98 | 10.91 0.36 9 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.867 0.362 10 1.00 | 149.7 5.4 10 0.99 | 0.4244 0.0143 10 1.00 | 54.01 2.25 10 1.00 | 19.06 0.93 10 0.98 | 352.6 4.3 10 0.99 | 10.68 0.35 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.744 0.368 10 0.99 | 144.3 5.5 10 0.95 | 0.4116 0.0156 10 0.97 | 53.20 1.94 10 0.98 | 18.64 0.75 10 0.96 | 350.5 6.8 10 0.98 | 10.82 0.33 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 833.0 141.8 9 - | 183.07 34.30 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 801.3 145.2 8 0.96 | 178.02 22.41 9 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 796.3 97.2 9 0.96 | 185.18 28.61 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 910.6 58.3 10 1.09 | 177.80 38.40 10 0.97 |
Table 21. Summary of Reproductive Performance
F0 Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 25 25 25 5.3 5.1 25 - 19 76.0 6 24.0 100.0 25/25 | 25 25 25 3.2 2.5 25 -39.4 24 96.0 1 4.0 100.0 25/25 | 25 25 25 2.7 2.6 25 -49.2 24 96.0 1 4.0 100.0 25/25 | 25 24 24 3.0 2.7 24 -43.2 23 95.8 1 4.2 100.0 24/24 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 22. Summary of Reproductive Performance
F1 Animals - Cohort 1B Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 20 20 20 2.7 1.0 20 - 20 100.0 0 0.0 100.0 20/20 | 20 20 20 3.9 3.7 20 45.3 18 90.0 2 10.0 100.0 20/20 | 20 20 20 3.5 2.7 20 32.1 19 95.0 1 5.0 100.0 20/20 | 20 20 20 2.5 1.2 20 -7.5 20 100.0 0 0.0 100.0 20/20 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 23. Mating Performance and Fertility
F0 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 6(1)* | 4 | 8 | 7(1) |
2 | 2 | 5 | 6 | 5 |
3 | 6 | 8 | 7 | 3 |
4 | 5 | 7 | 3 | 8 |
No clear indication of mating | 6(1) | 1 | 1 | 1 |
Mean number of nights to positive mating sign | 2.5 | 2.8 | 2.2 | 2.5 |
Number passing one estrus | 0 | 0 | 0 | 0 |
Number of males paired | 25 | 25 | 25 | 24 |
Number of siring males | 24 | 25 | 25 | 23 |
Male Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Male Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Male Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
Female Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Female Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Female Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
*Includes Animal 1524 that had 2 implant sites (no live or dead pups)
Table 24. Mating Performance and Fertility
F1 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 3 | 5 | 2 | 7 |
2 | 5 | 2 | 4 | 2 |
3 | 9 | 5(2) | 8 | 6 |
4 | 2 | 5 | 4 | 5 |
5 | 1(1) | 1 | 0 | 0 |
6 | 0 | 0 | 1 | 0 |
No clear indication of mating | 0 | 2(2) | 1(1) | 0 |
Mean number of nights to positive mating sign | 2.7 | 2.7 | 2.9 | 2.5 |
Number passing one estrus | 0 | 1 | 1 | 0 |
Number of males paired | 20 | 20 | 20 | 20 |
Number of siring males | 19 | 16 | 19 | 20 |
Male Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Male Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Male Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Female Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Female Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Female Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Table 25. Summary of Duration of Gestation and Overall Litter Performance
F0 Animals
F0 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 24 | 25 | 25 | 23 |
Duration of Gestation (Days) 21 |
1 |
1 |
0 |
0 |
22 | 17 | 17 | 23 | 20 |
23 | 0 | 6 | 1 | 2 |
No clear indication of mating | 6 | 1 | 1 | 1 |
Mean Duration | 21.9 | 22.2 | 22.0 | 22.1 |
Number of females producing a live litter | 23 | 25 | 25 | 23 |
Gestation index as % | 96 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.0 ± 3.4 | 12.5 ± 2.4 | 12.6 ± 1.9 | 12.9 ± 1.6 |
Mean total number of pups born per litter ± standard deviation | 11.0 ± 3.0 | 11.2 ± 2.3 | 11.0 ± 2.5 | 11.0 ± 3.3 |
Mean number of live pups per litter ± standard deviation: Lactation Day 1 |
11.0 ± 3.0 |
11.1 ± 2.2 |
10.9 ± 2.6 |
11.1 ± 2.6 |
Lactation Day 4 | 10.9 ± 3.1 | 11.2 ± 2.2 | 10.9 ± 2.6 | 11.0 ± 2.7 |
Lactation Day 4a | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 7 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8± 0.7 | 7.7 ± 0.9 |
Lactation Day 14 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 21 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Number of males on Lactation Day 1 (%) | 120 (47) | 135 (49) | 145 (53) | 116 (47) |
Number of females on Lactation Day 1 (%) | 134 (53) | 143 (51) | 127 (47) | 129 (53) |
a After cull of pups on PND 4 to standardise the size of the litter
Table 26. Summary of Duration of Gestation and Overall Litter Performance
F1 Animals
F1 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 19 | 16 | 19 | 20 |
Duration of Gestation (Days) 21 |
1 |
3 |
1 |
1 |
22 | 15 | 9 | 17 | 18 |
23 | 3 | 4 | 1 | 1 |
Mean Duration | 22.1 | 22.1 | 22.0 | 22.0 |
Number of females producing a live litter | 19 | 16 | 19 | 20 |
Gestation index as % | 100 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.6 ± 1.7 | 12.3 ± 2.3 | 13.0 ± 1.6 | 12.5 ± 2.3 |
Mean total number of pups born per litter ± standard deviation* | 11.6 ± 1.9 | 11.1 ± 2.8 | 10.5 ± 2.8 | 11.3 ± 2.2 |
Mean number of live pups per litter ± standard deviation*: Lactation Day 1 |
11.3 ± 2.2 |
11.1 ± 3.0 |
10.4 ± 2.7 |
11.1 ± 2.1 |
Lactation Day 4 | 11.3 ± 2.2 | 11.0 ± 2.9 | 10.3 ± 2.8 | 11.0 ± 2.1 |
Lactation Day 4a | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 7 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 14 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 21 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.8 ± 0.5 |
Number of males on Lactation Day 1 (%) | 96 (47) | 96 (54) | 93 (49) | 100 (45) |
Number of females on Lactation Day 1 (%) | 108 (53) | 81 (46) | 96 (51) | 121 (55) |
a After cull of pups on PND 4 to standardise the size of the litter
* Excludes Animal 1628 that had a total litter loss
Table 27. Summary of Urinalysis Values
F0 Animals - Males
Day: 131 Relative to Start Date
| Sex: Male | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G1] | |||
| 0 mg/kg/day | Mean | 4.32 | 1.0152 | 7.5 |
| Group 1 | SD | 2.94 | 0.0089 | 0.94 |
| N | 10 | 10 - | 10 - | |
| - | ||||
| 100 mg/kg/day | Mean | 5.64 | 1.0135 | 8 |
| Group 2 | SD | 2.99 | 0.0052 | 0.62 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.31 | 1 | 1.07 | |
| 300 mg/kg/day | Mean | 4.22 | 1.0215 | 8.15 |
| Group 3 | SD | 2.11 | 0.0086 | 0.63 |
| N | 10 | 10 | 10 | |
| tCtrl | 0.98 | 1.01 | 1.09 | |
| 900 mg/kg/day | Mean | 5.25 | 1.0300 ** | 7.35 0.71 |
| Group 4 | SD | 2.39 | 0.0133 | 10 |
| N | 10 | 10 | 0.98 | |
| tCtrl | 1.22 | 1.01 |
Table 28. Summary of Urinalysis Values
F0 Animals - Females
Day: 108 Relative to Start Date
| Sex: Female | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G1] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 4.85 | 1.0188 | 8.6 |
| Group 1 | SD | 2.33 | 0.0052 | 0.32 |
| N | 10 - | 10 - | 10 - | |
| 100 mg/kg/day | Mean | 5.23 | 1.0192 | 8.45 |
| Group 2 | SD | 2.96 | 0.0069 | 0.16 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.08 | 1 | 0.98 | |
| 300 mg/kg/day | Mean | 6.01 | 1.0199 | 8.2 |
| Group 3 | SD | 1.68 | 0.0039 | 0.42 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.24 | 1 | 0.95 | |
| 900 mg/kg/day | Mean | 7.05 2.58 | 1.0218 0.0051 | 8.05 * |
| Group 4 | SD | 10 | 10 | 0.72 |
| N | 1.45 | 1 | 10 | |
| tCtrl | 0.94 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05
Table 29. Summary of Urinalysis Values
F1 Animals - Cohort 1A
Day: 60 Relative to Start Date
| Sex: Male | Reporting urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 2.67 | 1.0182 | 8.11 |
| Group 1 | SD | 1.87 | 0.0078 | 0.6 |
| N | 10 - | 10 - | 9 - | |
| 100 mg/kg/day | Mean | 3.07 | 1.0168 | 7.9 |
| Group 2 | SD | 0.95 | 0.0123 | 0.61 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.15 | 1 | 0.97 | |
| 300 mg/kg/day | Mean | 1.96 | 1.0302 * | 7.11 ** 0.70 |
| Group 3 | SD | 1.25 | 0.0118 | 9 |
| N | 10 | 10 | 0.88 | |
| tCtrl | 0.73 | 1.01 | ||
| 900 mg/kg/day | Mean | 2.94 1.43 | 1.0346 ** | 6.25 ** |
| Group 4 | SD | 10 | 0.0091 | 0.42 |
| N | 1.1 | 10 | 10 | |
| tCtrl | 1.02 | 0.77 |
Day: 60 Relative to Start Date
Sex: Female |
|
| Reporting Urinalysi | s |
VOLUME (mL) [G] | SPECIFIC GRAVITY [G] | URINE pH [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.88 1.25 10 - | 1.0226 0.0117 10 - | 7.00 0.94 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 3.20 2.88 10 1.70 | 1.0161 0.0086 10 0.99 | 7.15 0.91 10 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.33 1.08 10 0.71 | 1.0333 0.0118 10 1.01 | 6.61 0.55 9 0.94 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.61 1.55 9 1.39 | 1.0267 0.0087 9 1.00 | 6.44 0.68 9 0.92 |
[G] - Anova & Dunnett
Table 1 - Summary of Body Weights (g)
F0 Males
| Sex: Male | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 124.9 | 176.6 | 225.1 | 268.9 | 303.1 | 333 | 355.5 |
| Group 1 | SD | 16.6 | 18.5 | 21 | 23.8 | 24.7 | 26.3 | 27.3 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 123.8 | 174.3 | 225.7 | 270.7 | 305.6 | 333.2 | 355.7 |
| Group 2 | SD | 16.3 | 18.8 | 23.4 | 25.2 | 26.4 | 28.5 | 30.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.9 | -1.3 | 0.3 | 0.7 | 0.8 | 0.1 | 0.1 | |
| 300 mg/kg/day | Mean | 121 | 171.9 | 222.1 | 268.3 | 302.8 | 332.4 | 355.9 |
| Group 3 | SD | 15.1 | 18.9 | 21 | 23.1 | 22.1 | 24.6 | 26.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -3.1 | -2.7 | -1.3 | -0.2 | -0.1 | -0.2 | 0.1 | |
| 900 mg/kg/day | Mean | 124.9 | 178.2 | 230.5 | 277.9 | 310.1 | 334.8 | 352.2 |
| Group 4 | SD | 11.6 | 14.2 | 18.9 | 21.9 | 24.1 | 25.7 | 28.3 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0 | 0.9 | 2.4 | 3.4 | 2.3 | 0.5 | -0.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 43 | 50 | 57 | 64 | 71 | 78 | 85 | ||
| 0 mg/kg/day | Mean | 374.2 | 389 | 406.9 | 417.8 | 427.7 | 428 | 435.6 |
| Group 1 | SD | 30.2 | 31.6 | 33.7 | 35 | 34.9 | 33.2 | 34.4 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 374.4 | 392.8 | 408 | 417.3 | 423.9 | 427.8 | 437.4 |
| Group 2 | SD | 32.8 | 34.9 | 36.1 | 35.5 | 36.7 | 37.4 | 37.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 0.1 | 1 | 0.3 | -0.1 | -0.9 | -0.1 | 0.4 | |
| 300 mg/kg/day | Mean | 373.7 | 390.5 | 404.8 | 415.6 | 424 | 426.6 | 436.5 |
| Group 3 | SD | 29.8 | 32.2 | 33.2 | 33.8 | 36.9 | 36.4 | 38.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | 0.4 | -0.5 | -0.5 | -0.9 | -0.3 | 0.2 | |
| 900 mg/kg/day | Mean | 365.9 | 378.8 | 393.4 | 403.8 | 411.4 | 415.7 | 423.1 |
| Group 4 | SD | 29.7 | 32 | 33.1 | 34.7 | 35.5 | 34.6 | 36.7 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -2.2 | -2.6 | -3.3 | -3.3 | -3.8 | -2.9 | -2.9 | |
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 92 | 99 | 106 | 113 | 120 | 127 | 134 | ||
| 0 mg/kg/day | Mean | 441.8 | 453.6 | 466.3 | 472.8 | 483.5 | 489.2 | 496.7 |
| Group 1 | SD | 35.9 | 36.1 | 37.8 | 37.8 | 38.9 | 39 | 41.7 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 441.6 | 450.2 | 462.2 | 470.4 | 478 | 487.7 | 492.6 |
| Group 2 | SD | 37.1 | 37.6 | 39.7 | 40.7 | 41.1 | 43.8 | 45.4 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -0.1 | -0.7 | -0.9 | -0.5 | -1.1 | -0.3 | -0.8 | |
| 300 mg/kg/day | Mean | 446.7 | 452.6 | 463.6 | 475.6 | 483.9 | 492.7 | 496.6 |
| Group 3 | SD | 40.7 | 42.3 | 44.7 | 47 | 49.7 | 50.9 | 53.1 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 1.1 | -0.2 | -0.6 | 0.6 | 0.1 | 0.7 | 0 | |
| 900 mg/kg/day | Mean | 434.1 | 442.2 | 449.5 | 458.4 | 464.4 | 475.5 | 478.6 |
| Group 4 | SD | 38 | 39.8 | 40.3 | 40.7 | 40.7 | 42.5 | 44.8 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | -1.7 | -2.5 | -3.6 | -3 | -3.9 | -2.8 | -3.6 | |
| Sex: Male | Day(s) Relative to Start Date | ||||
| 137 | 138 | 141 | 142 | ||
| 0 mg/kg/day | Mean | 507.9 | 501.8 | 493.1 | 473.4 |
| Group 1 | SD | 58.2 | 33.5 | 36.1 | 43.3 |
| N | 8 - | 6 - | 11 - | 5 - | |
| 100 mg/kg/day | Mean | 488 | 512.6 | 490.8 | 477.7 |
| Group 2 | SD | 46.5 | 39.6 | 50.5 | 63.9 |
| N | 5 | 7 | 13 | 7 | |
| %Diff | -3.9 | 2.1 | -0.5 | 0.9 | |
| 300 mg/kg/day | Mean | 458 | 534.4 | 491 | 494.8 |
| Group 3 | SD | 26.4 | 57.8 | 46.4 | 52.9 |
| N | 4 | 8 | 13 | 8 | |
| %Diff | -9.8 | 6.5 | -0.4 | 4.5 | |
| 900 mg/kg/day | Mean | 473.8 | 494.5 | 480.2 | 471.2 |
| Group 4 | SD | 41.8 | 27.5 | 54.1 | 65.1 |
| N | 8 | 4 | 13 | 5 | |
| %Diff | -6.7 | -1.5 | -2.6 | -0.5 | |
Table 2 - Summary of Body Weights (g)
F0 Females - Prior to mating
| Sex: Female | Day(s) Relative to Start Date | |||||||
| -7 | 1 | 8 | 15 | 22 | 29 | 36 | ||
| 0 mg/kg/day | Mean | 72.9 | 110.5 | 136.4 | 155.4 | 173.6 | 188.9 | 197.4 |
| Group 1 | SD | 13.5 | 13.9 | 14 | 13.8 | 13.3 | 13.9 | 14.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 76 | 114.9 | 140.8 | 162 | 181.1 | 195.7 | 206.3 |
| Group 2 | SD | 10.2 | 11.1 | 11.6 | 13.5 | 14.4 | 14.5 | 16.5 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 3.9 | 3.3 | 4.3 | 4.3 | 3.6 | 4.5 | |
| 300 mg/kg/day | Mean | 74.9 | 114.2 | 142.2 | 162.9 | 183.5 | 198.8 | 209.1* |
| Group 3 | SD | 12.4 | 11.1 | 10.6 | 12.3 | 12.8 | 14.7 | 15.6 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 2.8 | 3.4 | 4.3 | 4.9 | 5.7 | 5.2 | 5.9 | |
| 900 mg/kg/day | Mean | 77.2 | 116.4 | 147.2* | 171.3** | 191.6** | 208.2** | 214.9** |
| Group 4 | SD | 14.1 | 14.3 | 15.3 | 17.2 | 17.4 | 17.9 | 18.2 |
| N | 25 | 25 | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 5.9 | 5.3 | 7.9 | 10.3 | 10.3 | 10.2 | 8.9 |
| Sex: Female | Day(s) Relative to Start Date | |||||
| 43 | 50 | 57 | 64 | 71 | ||
| 0 mg/kg/day | Mean | 207.3 | 214.6 | 222.2 | 226.2 | 230.5 |
| Group 1 | SD | 14.9 | 13.1 | 14.8 | 14.4 | 15.6 |
| N | 25 - | 25 - | 25 - | 25 - | 25 - | |
| 100 mg/kg/day | Mean | 216 | 225.6 | 229.6 | 232.8 | 240.8 |
| Group 2 | SD | 17.1 | 18.4 | 19 | 20.5 | 20.5 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.2 | 5.2 | 3.3 | 2.9 | 4.5 | |
| 300 mg/kg/day | Mean | 217.2 | 227.9* | 234.7* | 238.4 | 245.9* |
| Group 3 | SD | 16.5 | 18.6 | 19.4 | 18.9 | 19 |
| N | 25 | 25 | 25 | 25 | 25 | |
| %Diff | 4.8 | 6.2 | 5.6 | 5.4 | 6.7 | |
| 900 mg/kg/day | Mean | 227.0** | 233.0** | 242.0** | 244.5** | 250.5** |
| Group 4 | SD | 17.2 | 17.5 | 18.6 | 19 | 19.7 |
| N | 24 | 24 | 24 | 24 | 24 | |
| %Diff | 9.5 | 8.6 | 8.9 | 8.1 | 8.7 |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 3 - Summary of Body Weights (g)
F0 Females - Gestation
| Sex: Female | Day(s) Relative to Mating (Litter: A) | ||||
| 0 | 7 | 14 | 20 | ||
| 0 mg/kg/day | Mean | 222.8 | 246.1 | 269.9 | 325.9 |
| Group 1 | SD | 14.5 | 13.5 | 16.3 | 24 |
| N | 18 - | 18 - | 18 - | 18 - | |
| 100 mg/kg/day | Mean | 237.2* | 259.5 | 282.3 | 338.5 |
| Group 2 | SD | 20.1 | 20.5 | 22.9 | 31.2 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 6.5 | 5.5 | 4.6 | 3.9 | |
| 300 mg/kg/day | Mean | 242.1** | 262.4* | 284.2 | 343.6 |
| Group 3 | SD | 20.5 | 22.4 | 24.6 | 28.1 |
| N | 24 | 24 | 24 | 24 | |
| %Diff | 8.6 | 6.6 | 5.3 | 5.4 | |
| 900 mg/kg/day | Mean | 243.7** | 264.5* | 291.1* | 352.5* |
| Group 4 | SD | 17.2 | 21.1 | 23.4 | 34.4 |
| N | 22 | 22 | 22 | 22 | |
| %Diff | 9.4 | 7.5 | 7.9 | 8.2 | |
Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 4 - Summary of Body Weights (g)
F0 Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | ||
| Sex | 7 | ||||||
| 1F | Mean | 261.7 | 274 | 275.1 | 282.8 | 280.1 | |
| SD | 19.1 | 20.8 | 19.3 | 21.2 | 16.6 | ||
| N | 23 | 22 | 23 | 23 | 23 | ||
| Mean | 268.2 | 280.9 | 283.3 | 290.9 | 288 | ||
| 2F | SD | 22.1 | 22.6 | 23 | 23.6 | 21.7 | |
| N | 23 | 24 | 24 | 24 | 24 | ||
| %Diff G1 | 2.5 | 2.5 | 3 | 2.9 | 2.8 | ||
| Mean | 270.8 | 282.7 | 285.6 | 294.8 | 291 | ||
| SD | 25.7 | 25 | 24.4 | 23.4 | 19.9 | ||
| 3F | N | 25 | 25 | 25 | 25 | 25 | |
| %Diff G1 | 3.5 | 3.2 | 3.8 | 4.2 | 3.9 | ||
| Mean | 271.6 | 286.4 | 292.2 | 302.6a | 300.1b | ||
| 4F | SD | 23.4 | 24 | 24.8 | 25.7 | 25.2 | |
| N | 22 | 22 | 22 | 22 | 22 | ||
| %Diff G1 | 3.8 | 4.5 | 6.2 | 7 | 7.1 | ||
Significantly different from control group 1 value :a=p≤0.05,b=p≤0.01 (Dunnett)
Table 5 - Summary of Body Weights (g)
F1 Animals - Cohort 1A
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
| 0 mg/kg/day | Mean | 56 | - | 71.8 | 119.9 | 172.1 | 219.5 | 269.6 |
| Group 1 | SD | 5.1 | - | 8.8 | 12.2 | 15.8 | 19.7 | 23.4 |
| N | 20 - | - | 20 - | 20 - | 20 - | 20 - | 20 - | |
| - | ||||||||
| 100 mg/kg/day | Mean | 56.3 | 61.0n | 70.8 | 116.4 | 166.9 | 213.7 | 261.2 |
| Group 2 | SD | 6.6 | 6.6 | 9 | 13.6 | 16.5 | 20.5 | 26.4 |
| N | 19 | 3 - | 18 | 20 | 20 | 20 | 20 | |
| %Diff | 0.7 | -1.4 | -2.9 | -3 | -2.6 | -3.1 | ||
| 300 mg/kg/day | Mean | 57.9 | 76.0n | 76.8 | 126.3 | 179.8 | 227 | 276.8 |
| Group 3 | SD | 4.9 | - | 9.3 | 14 | 16.9 | 19.6 | 24.9 |
| N | 20 | 1 - | 19 | 20 | 20 | 20 | 20 | |
| %Diff | 3.5 | 6.9 | 5.4 | 4.5 | 3.4 | 2.7 | ||
| 900 mg/kg/day | Mean | 54.7 | 59.0n 4.3 | 70.6 | 115.8 | 167.5 | 212.9 | 258.5 |
| Group 4 | SD | 5.5 | 6 - | 8.2 | 12.5 | 16.5 | 21.7 | 25.5 |
| N | 15 | 19 | 20 | 20 | 20 | 20 | ||
| %Diff | -2.2 | -1.6 | -3.4 | -2.7 | -3 | -4.1 |
[G] - Anova & Dunnett [I] - n - Inappropriate for statistics
| Sex: Male | Day(s) Relative to Start Date | |||||||
| 41 | 48 | 55 | 62 | 68 | 69 | 70 | ||
| 0 mg/kg/day | Mean | 309.2 | 343.6 | 368.2 | 390.7 | 408.1 | 394 | 405.5 |
| Group 1 | SD | 26.7 | 31.5 | 33 | 37 | 40.1 | 42.2 | 40.7 |
| N | 20 - | 20 - | 20 - | 20 - | 14 - | 6 - | 20 - | |
| 100 mg/kg/day | Mean | 300.1 | 335.3 | 360.5 | 382.8 | 393 | 408.2 | 401.6 |
| Group 2 | SD | 29.4 | 32.6 | 35.7 | 36.5 | 35.3 | 42.7 | 34.3 |
| N | 20 | 20 | 20 | 20 | 16 | 6 | 16 | |
| %Diff | -2.9 | -2.4 | -2.1 | -2 | -3.7 | 3.6 | -1 | |
| 300 mg/kg/day | Mean | 317.3 | 349.4 | 376.1 | 396.8 | 411.1 | 445.9 | 411.1 |
| Group 3 | SD | 28 | 33 | 37.6 | 38.7 | 46.2 | 36.6 | 36.1 |
| N | 20 | 20 | 20 | 20 | 14 | 7 | 19 | |
| %Diff | 2.6 | 1.7 | 2.1 | 1.6 | 0.8 | 13.2 | 1.4 | |
| 900 mg/kg/day | Mean | 299.2 | 325.5 | 346.4 | 363.5 | 386.8 | 381.7 | 382.7 |
| Group 4 | SD | 29 | 32.1 | 37.5 | 40 | 40.6 | 48.2 | 38.9 |
| N | 20 | 20 | 20 | 20 | 14 | 11 | 15 | |
| %Diff | -3.2 | -5.3 | -5.9 | -7 | -5.2 | -3.1 | -5.6 |
Sex: Male |
| Day(s) Relative to Start Date |
71 | ||
0 mg/kg/day
Group 1 | Mean SD N
| - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 447.0n 69.3 2 - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | - - - - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | - - - - |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 4.9 20 - | - - - - | 66.0 7.8 20 - | 103.8 9.7 20 - | 136.5 10.7 20 - | 157.2 10.2 20 - | 176.5 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 55.3 4.9 19 5.3 | 58.0n 3.0 3 - | 68.9 5.7 18 4.5 | 106.1 9.4 20 2.2 | 138.3 11.9 20 1.3 | 159.7 14.1 20 1.6 | 178.8 16.8 20 1.3 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.0 5.0 21 4.7 | 71.0n - 1 - | 69.4 8.9 19 5.2 | 105.5 11.3 20 1.7 | 138.6 11.6 20 1.5 | 160.2 12.3 20 1.9 | 179.9 13.8 20 1.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 52.1 4.8 15 -0.8 | 56.3n 6.0 7 - | 66.1 8.2 18 0.2 | 103.2 10.6 20 -0.6 | 138.6 12.8 20 1.5 | 159.9 15.9 20 1.7 | 182.0 15.5 20 3.1 |
[G] - Anova & Dunnett
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 [G] | 48 [G] | 55 [G] | 62 [G] | 68 [G] | 69 [G] | 70 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.0 12.1 20 - | 203.2 14.4 20 - | 212.2 15.5 20 - | 221.0 18.8 20 - | 231.6 18.8 13 - | 228.1 18.9 7 - | 245.3n 11.0 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 192.4 18.5 20 1.8 | 205.7 17.3 20 1.2 | 214.1 18.0 20 0.9 | 224.2 20.5 20 1.4 | 235.2 22.6 17 1.6 | 216.7 6.8 3 -5.0 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 194.4 13.6 20 2.8 | 206.5 14.6 20 1.6 | 216.4 16.9 20 2.0 | 226.1 17.7 20 2.3 | 231.5 16.3 14 0.0 | 242.3 12.6 6 6.2 | 220.0n - 1 -10.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 201.6 17.1 20 6.7 | 209.2 17.6 20 3.0 | 218.0 17.7 20 2.7 | 226.5 18.3 20 2.5 | 237.5 19.1 15 2.6 | 235.6 26.0 5 3.3 | - - - - |
[G] - Anova & Dunnett
Sex: Female |
| Day(s) Relative to Start Date |
| ||
71 [G] | 72 [I] | 73 [G] | 74 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 226.9 16.6 8 - | - - - - | 228.6 21.1 5 - | 228.8 17.9 4 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 228.3 29.4 8 0.6 | 223.7n 12.2 3 - | 233.8 11.4 5 2.3 | 233.8 25.4 4 2.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 233.1 15.5 9 2.7 | 268.0n - 1 - | 225.0n 11.3 2 -1.6 | 237.7 20.5 7 3.9 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 235.0 23.9 10 3.6 | 247.0n 11.3 2 - | 232.8 19.2 6 1.9 | 235.0n 25.5 2 2.7 |
[G] - Anova & Dunnett: n - Inappropriate for statistics
Table 7 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Males
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 5.4 20 - | - - - - | 70.5 8.5 20 - | 116.5 12.0 20 - | 167.7 13.3 20 - | 212.9 17.3 20 - | 261.7 20.9 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 57.4 5.2 19 5.1 | 60.0n 8.5 3 - | 72.9 8.7 18 3.5 | 117.5 14.9 20 0.8 | 166.8 20.5 20 -0.6 | 211.2 25.9 20 -0.8 | 258.5 29.7 20 -1.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 56.8 5.7 20 4.0 | 70.0n - 1 - | 74.9 10.8 19 6.2 | 124.1 15.9 20 6.5 | 175.7 18.3 20 4.7 | 222.6 21.4 20 4.6 | 271.2 25.8 20 3.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 54.5 4.5 15 -0.1 | 59.3n 5.9 7 - | 69.9 8.7 18 -0.8 | 115.4 10.8 20 -0.9 | 166.8 12.3 20 -0.5 | 212.2 15.2 20 -0.3 | 258.6 18.7 20 -1.2 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 300.5 25.6 20 - | 334.6 30.5 20 - | 358.4 34.8 20 - | 381.5 38.4 20 - | 395.5 41.5 20 - | 410.3 44.0 20 - | 421.2 46.7 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 297.5 32.5 20 -1.0 | 330.9 34.9 20 -1.1 | 357.3 37.9 20 -0.3 | 379.6 39.8 20 -0.5 | 399.5 39.4 20 1.0 | 414.5 41.7 20 1.0 | 429.9 42.0 20 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 310.2 26.4 20 3.2 | 342.6 29.7 20 2.4 | 370.5 31.5 20 3.4 | 393.0 33.5 20 3.0 | 413.3 33.9 20 4.5 | 427.1 37.4 20 4.1 | 438.9 39.0 20 4.2 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 297.9 23.2 20 -0.8 | 324.7 26.1 20 -2.9 | 348.3 30.2 20 -2.8 | 365.3 33.5 20 -4.2 | 383.9 36.0 20 -2.9 | 399.6 35.5 20 -2.6 | 410.4 40.6 20 -2.6 |
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
90 | 97 | 104 | 111 | 118 | 125 | 128 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 431.7 46.1 20 - | 434.7 44.9 20 - | 441.9 43.5 20 - | 453.5 48.3 20 - | 464.3 49.9 20 - | 473.8 52.6 20 - | 484.1 58.9 8 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 440.9 44.6 20 2.1 | 444.3 45.0 20 2.2 | 452.5 46.6 20 2.4 | 463.5 46.1 20 2.2 | 477.7 47.1 20 2.9 | 492.3 48.2 20 3.9 | 494.2 58.5 11 2.1 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 448.1 41.2 20 3.8 | 452.0 38.8 20 4.0 | 456.2 39.7 20 3.2 | 471.1 40.7 20 3.9 | 483.8 43.3 20 4.2 | 494.5 45.3 20 4.4 | 515.1 42.9 9 6.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 415.5 40.8 20 -3.8 | 424.9 41.7 20 -2.3 | 428.4 41.3 20 -3.1 | 442.0 42.1 20 -2.5 | 451.7 42.8 20 -2.7 | 460.8 41.2 20 -2.8 | 463.5 45.3 12 -4.3 |
Sex: Male |
|
| Day(s) Relative to Start Date |
|
129 [G] | 130 [G] | 131 [I] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 465.0 62.2 6 - | 475.3 45.0 9 - | - - - - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 504.9 36.5 7 8.6 | 502.0 45.0 7 5.6 | - - - - |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 514.8 41.9 5 10.7 | 471.0 41.8 8 -0.9 | 485.5n 36.8 4 - |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 476.3 43.3 8 2.4 | 436.3 10.1 3 -8.2 | 529.0n - 1 - |
Table 8 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Prior to Mating
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 52.6 5.1 20 - | - - - - | 66.2 7.1 20 - | 103.6 9.0 20 - | 136.1 9.5 20 - | 156.9 10.6 20 - | 174.8 12.3 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 53.8 7.9 19 2.4 | 57.3n 4.0 3 - | 66.6 10.4 18 0.7 | 103.4 13.9 20 -0.1 | 133.7 13.8 20 -1.7 | 155.3 14.0 20 -1.0 | 173.5 14.1 20 -0.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 4.4 20 6.0 | 68.0n - 1 - | 71.6 7.5 19 8.3 | 109.3 8.1 20 5.5 | 143.2 9.6 20 5.3 | 164.1 10.0 20 4.6 | 180.9 11.8 20 3.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 51.3 4.3 15 -2.3 | 56.3n 6.6 7 - | 64.6 7.4 18 -2.4 | 102.8 10.3 20 -0.7 | 136.6 9.8 20 0.4 | 157.6 13.1 20 0.4 | 176.8 13.7 20 1.1 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
41 | 48 | 55 | 62 | 69 | 76 | 83 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 189.6 11.3 20 - | 204.2 11.0 20 - | 210.9 11.8 20 - | 218.6 12.6 20 - | 227.6 11.8 20 - | 232.5 11.5 20 - | 236.2 12.5 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 188.1 15.8 20 -0.8 | 200.3 15.1 20 -1.9 | 209.8 15.4 20 -0.5 | 218.7 14.7 20 0.1 | 226.3 16.5 20 -0.6 | 230.7 18.2 20 -0.8 | 232.7 17.3 20 -1.5 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 199.2 12.4 20 5.1 | 211.3 13.2 20 3.5 | 220.1 14.6 20 4.4 | 227.0 15.2 20 3.9 | 236.9 15.5 20 4.1 | 243.1 16.4 20 4.5 | 246.7 17.5 20 4.5 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 195.6 14.1 20 3.2 | 203.7 14.7 20 -0.2 | 212.2 15.4 20 0.6 | 219.5 16.9 20 0.4 | 231.0 16.5 20 1.5 | 236.4 18.1 20 1.7 | 237.4 17.1 20 0.5 |
Sex: Female |
| Day(s) Relative to Start Date |
90 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 240.0 13.4 20 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 239.6 16.7 20 -0.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 251.2 17.1 20 4.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 242.7 18.8 20 1.1 |
Table 9 - Summary of Body Weights (g)
F1 Animals - Cohort 1B - Gestation
Sex: Female |
|
| Day(s) Relative to Mating (Litter: A) |
| |
0 | 7 | 14 | 20 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 241.9 12.7 19 - | 261.8 15.3 19 - | 284.1 15.0 19 - | 337.5 18.9 19 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 238.8 14.3 16 -1.3 | 258.2 14.3 16 -1.4 | 277.9 14.9 16 -2.2 | 334.6 19.9 16 -0.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 253.5 16.6 19 4.8 | 271.6 16.9 19 3.8 | 291.6 16.6 19 2.6 | 348.9 19.3 19 3.4 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 241.1 16.7 20 -0.3 | 260.8 20.7 20 -0.4 | 278.0 23.3 20 -2.1 | 336.5 32.1 20 -0.3 |
Table 10 - Summary of Body Weights (g)
F1 Animals - Cohort 1B Females - Lactation
| Group / | 1 | 4 | Day | 14 | 21 | |
| Sex | 7 | |||||
| 1F | Mean | 264.2 | 268.9 | 276.9 | 290.6 | 284.2 |
| SD | 14.4 | 12.9 | 14.9 | 16.8 | 17.9 | |
| N | 18 | 17 | 18 | 18 | 18 | |
| Mean | 262.3 | 271.3 | 275.8 | 288.6 | 286.3 | |
| 2F | SD | 14.5 | 15 | 14.2 | 16.6 | 15.4 |
| N | 16 | 15 | 16 | 16 | 16 | |
| %Diff G1 | -0.7 | 0.9 | -0.4 | -0.7 | 0.8 | |
| Mean | 278.4a | 288.5b | 292.6a | 306.2a | 302.1a | |
| SD | 18.9 | 16.6 | 15.7 | 15.6 | 16.2 | |
| 3F | N | 19 | 19 | 19 | 19 | 19 |
| %Diff G1 | 5.4 | 7.3 | 5.6 | 5.4 | 6.3 | |
| Mean | 265.9 | 276.3 | 283.6 | 292.4 | 291.5 | |
| 4F | SD | 21.6 | 24.5 | 20.3 | 21.7 | 22.9 |
| N | 20 | 19 | 20 | 20 | 19 | |
| %Diff G1 | 0.6 | 2.7 | 2.4 | 0.6 | 2.6 |
Table 11 - Summary of Body Weights (g)
F1 Animals - Cohort 2A
Sex: Male |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 54.6 4.7 10 - | - - - - | 68.4 8.0 10 - | 115.1 11.9 10 - | 169.5 12.1 10 - | 219.4 13.6 10 - | 269.5 18.2 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.6 5.5 9 -0.1 | 59.0n 7.1 2 - | 67.0 8.9 9 -2.0 | 111.0 12.0 10 -3.6 | 160.7 12.0 10 -5.2 | 205.3 14.3 10 -6.4 | 250.8 18.2 10 -6.9 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 58.5 5.9 10 7.1 | 76.0n - 1 - | 74.7 10.9 9 9.2 | 122.9 14.3 10 6.8 | 177.1 16.3 10 4.5 | 224.4 19.6 10 2.3 | 273.9 23.0 10 1.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 55.4 3.6 9 1.5 | 59.0n 7.1 2 - | 67.3 9.2 9 -1.6 | 112.4 11.1 10 -2.3 | 164.3 12.4 10 -3.1 | 211.4 15.1 10 -3.6 | 259.5 21.4 10 -3.7 |
Sex: Male |
|
|
| Day(s) Relative to Start Date |
|
| |
41 | 48 | 55 | 58 | 59 | 60 | ||
0 mg/kg/day
Group 1 | Mean SD N
| 308.0 21.9 10 - | 342.0 24.0 10 - | 368.4 25.7 10 - | 369.8 24.4 4 - | 396.8 32.0 5 - | 405.0 37.8 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 287.9 19.6 10 -6.5 | 320.4 24.2 10 -6.3 | 347.9 26.8 10 -5.6 | 376.8 42.4 4 1.9 | 369.8 20.2 6 -6.8 | 360.5 16.2 4 -11.0 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 313.5 26.9 10 1.8 | 348.1 27.9 10 1.8 | 372.8 25.9 10 1.2 | 403.3 32.3 6 9.1 | 404.0 54.1 3 1.8 | 369.5 14.5 4 -8.8 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 298.6 22.8 10 -3.1 | 326.5 27.2 10 -4.5 | 347.3 31.0 10 -5.7 | 362.0 46.0 4 -2.1 | 376.9 25.2 7 -5.0 | 341.5n 0.7 2 -15.7 |
Sex: Female |
|
|
|
| Day(s) Relative to Start Date |
|
|
|
1 [G] | 2 [I] | 6 [G] | 13 [G] | 20 [G] | 27 [G] | 34 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 51.5 4.2 10 - | - - - - | 62.9 6.5 10 - | 98.9 9.8 10 - | 128.6 9.3 10 - | 149.3 8.6 10 - | 166.7 9.1 10 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 54.2 2.9 9 5.3 | 54.0n 2.8 2 - | 65.4 5.5 9 4.0 | 100.5 8.6 10 1.6 | 131.3 8.2 10 2.1 | 155.1 8.6 10 3.9 | 172.1 10.8 10 3.2 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 55.7 7.4 10 8.2 | 76.0n - 1 - | 68.8 10.0 9 9.3 | 108.9 13.2 10 10.1 | 145.1** 12.9 10 12.8 | 167.6** 15.5 10 12.3 | 186.0** 18.3 10 11.6 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 53.6 3.7 9 4.0 | 57.0n 5.7 2 - | 63.2 5.3 9 0.5 | 100.5 6.2 10 1.6 | 135.7 10.3 10 5.5 | 159.2 10.4 10 6.6 | 179.3 12.7 10 7.6 |
Sex: Female |
|
| Day(s) Relative to Start Date |
|
| ||
41 [G] | 48 [G] | 55 [G] | 58 [G] | 59 [G1] | 60 [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 179.8 10.1 10 - | 193.9 9.9 10 - | 203.7 10.7 10 - | 204.8 13.1 4 - | 214.0 12.7 5 - | 214.3 12.2 3 - |
100 mg/kg/day
Group 2 | Mean SD N %Diff | 185.7 12.3 10 3.3 | 199.7 13.1 10 3.0 | 210.3 14.1 10 3.2 | 206.8 12.0 4 1.0 | 218.0 20.1 6 1.9 | 210.8 12.0 4 -1.7 |
300 mg/kg/day
Group 3 | Mean SD N %Diff | 202.3** 20.4 10 12.5 | 215.0** 20.3 10 10.9 | 223.8* 23.4 10 9.9 | 237.2 33.5 6 15.8 | 247.0 47.1 3 15.4 | 223.5 14.2 4 4.3 |
900 mg/kg/day
Group 4 | Mean SD N %Diff | 198.6* 14.7 10 10.5 | 210.7* 14.0 10 8.7 | 218.6 13.4 10 7.3 | 223.0 22.0 4 8.9 | 229.9 10.5 7 7.4 | 220.7 6.0 3 3.0 |
Tables 12. Summary of Clinical Chemistry Values
F0 Animals - Males. Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G1] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 71.4 12.3 10 - | 51.0 11.7 10 - | 128.5 12.4 10 - | 1.5 0.0 10 - | 217.1 68.4 10 - | 1.25 0.00 10 - | 7.35 0.66 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 74.6 10.3 9 1.04 | 49.1 12.0 9 0.96 | 126.0 25.8 9 0.98 | 1.5 0.0 9 1.00 | 292.7 163.0 9 1.35 | 1.25 0.00 9 1.00 | 6.90 0.68 9 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 60.5 * 4.9 10 0.85 | 44.5 7.8 10 0.87 | 125.1 32.5 10 0.97 | 1.5 0.0 10 1.00 | 213.2 70.8 10 0.98 | 1.25 0.00 10 1.00 | 7.16 0.35 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.6 * 5.4 10 0.86 | 36.8 14.1 9 0.72 | 115.4 28.5 10 0.90 | 1.5 0.0 10 1.00 | 196.8 60.0 10 0.91 | 1.25 0.00 10 1.00 | 6.94 0.68 10 0.94 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.2 3.2 10 - | 8.251 0.699 10 - | 1.789 0.249 10 - | 1.949 0.657 10 - | 66.29 1.89 10 - | 40.36 1.53 10 - | 25.93 2.18 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 32.1 3.5 9 0.94 | 8.778 1.195 9 1.06 | 1.853 0.393 9 1.04 | 1.933 0.681 9 0.99 | 66.09 3.31 9 1.00 | 41.22 2.83 9 1.02 | 24.87 2.61 9 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 * 2.9 10 0.87 | 8.113 0.521 10 0.98 | 2.170 * 0.404 10 1.21 | 1.594 0.199 10 0.82 | 66.67 2.46 10 1.01 | 41.93 1.91 10 1.04 | 24.74 2.11 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 31.5 3.6 10 0.92 | 8.208 0.634 10 0.99 | 2.327 ** 0.270 10 1.30 | 1.457 0.567 10 0.75 | 68.21 2.50 10 1.03 | 43.17 * 2.11 10 1.07 | 25.04 2.86 10 0.97 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.58 0.18 10 - | 2.618 0.059 10 - | 141.1 1.3 10 - | 4.98 0.29 10 - | 103.7 1.2 10 - | 1.404 0.193 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.69 0.22 9 1.07 | 2.637 0.076 9 1.01 | 141.4 1.9 9 1.00 | 4.90 0.34 9 0.98 | 103.7 1.4 9 1.00 | 1.531 0.178 9 1.09 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.71 0.17 10 1.08 | 2.654 0.055 10 1.01 | 140.3 2.2 10 0.99 | 4.97 0.15 10 1.00 | 102.3 1.8 10 0.99 | 1.545 0.152 10 1.10 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.75 0.28 10 1.11 | 2.716 ** 0.069 10 1.04 | 141.3 1.7 10 1.00 | 5.02 0.29 10 1.01 | 101.4 ** 2.0 10 0.98 | 1.591 0.187 10 1.13 |
Tables 13. Summary of Clinical Chemistry Values
F0 Animals - Females. Day: 115 Relative to Start Date
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 131.2 28.7 9 - | 117.1 17.5 9 - | 223.4 49.2 9 - | 1.5 0.0 9 - | 630.0 384.5 9 - | 1.25 0.00 9 - | 10.50 1.01 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 105.3 10.9 8 0.80 | 110.0 18.4 8 0.94 | 200.3 73.8 7 0.90 | 1.5 0.0 8 1.00 | 356.8 132.6 8 0.57 | 1.25 0.00 8 1.00 | 10.11 0.94 8 0.96 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 116.1 21.8 9 0.88 | 105.7 14.1 9 0.90 | 157.1 38.5 9 0.70 | 1.5 0.0 9 1.00 | 614.7 356.8 9 0.98 | 1.25 0.00 9 1.00 | 9.38 0.84 9 0.89 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 106.4 25.3 10 0.81 | 100.4 21.0 10 0.86 | 173.2 49.5 10 0.78 | 1.5 0.0 9 1.00 | 428.3 244.8 9 0.68 | 1.25 0.00 9 1.00 | 10.57 1.26 9 1.01 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G1] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 29.4 3.2 9 - | 4.740 1.150 9 - | 2.828 0.449 9 - | 0.589 0.061 9 - | 56.30 2.58 9 - | 40.21 1.79 9 - | 16.09 2.22 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 24.6 * 4.2 8 0.84 | 5.008 0.434 8 1.06 | 2.641 0.493 8 0.93 | 0.461 0.140 8 0.78 | 55.21 2.41 8 0.98 | 39.51 2.62 8 0.98 | 15.70 0.72 8 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 24.0 * 2.5 8 0.82 | 4.863 0.808 8 1.03 | 2.889 0.655 9 1.02 | 0.535 0.177 8 0.91 | 56.38 2.85 8 1.00 | 40.59 2.45 9 1.01 | 16.19 1.63 8 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 25.6 4.6 9 0.87 | 5.003 0.815 9 1.06 | 2.827 0.925 9 1.00 | 0.479 0.115 9 0.81 | 58.48 3.36 9 1.04 | 42.81 2.93 10 1.06 | 15.68 1.98 9 0.97 |
Sex: Female |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.53 0.34 9 - | 2.680 0.100 9 - | 140.7 2.1 9 - | 5.12 0.42 9 - | 99.8 2.2 9 - | 1.708 0.538 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.54 0.23 8 1.00 | 2.621 0.139 8 0.98 | 140.0 3.5 8 1.00 | 5.05 0.57 8 0.99 | 100.1 1.9 8 1.00 | 1.649 0.518 8 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.50 0.29 8 0.99 | 2.694 0.096 9 1.01 | 140.1 1.3 9 1.00 | 5.24 0.62 9 1.02 | 99.3 1.7 9 1.00 | 1.538 0.510 8 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.78 0.47 9 1.10 | 2.781 0.183 9 1.04 | 140.1 2.3 10 1.00 | 5.19 0.42 10 1.01 | 97.8 2.9 10 0.98 | 1.949 0.766 9 1.14 |
Tables 14. Summary of Clinical Chemistry Values
F1 Animals - Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 68.5 6.0 10 - | 57.6 11.4 10 - | 153.4 23.5 10 - | 1.5 0.0 9 - | 239.9 42.0 9 - | 1.25 0.00 9 - | 7.02 0.68 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 63.0 7.2 10 0.92 | 52.0 7.9 10 0.90 | 153.5 23.8 10 1.00 | 1.5 0.0 10 1.00 | 239.2 101.7 10 1.00 | 1.25 0.00 10 1.00 | 6.32 0.62 10 0.90 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 61.3 * 5.3 10 0.89 | 49.9 8.6 10 0.87 | 143.2 27.9 10 0.93 | 1.5 0.0 10 1.00 | 210.5 55.4 10 0.88 | 1.25 0.00 10 1.00 | 6.84 0.59 10 0.97 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 61.2 * 4.0 10 0.89 | 42.5 ** 13.0 10 0.74 | 141.5 23.9 10 0.92 | 1.5 0.0 10 1.00 | 174.9 37.4 10 0.73 | 1.25 0.00 10 1.00 | 7.24 0.85 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Biochemistry |
|
|
| |
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 30.8 4.0 9 - | 9.268 0.551 9 - | 1.723 0.237 9 - | 2.458 1.001 9 - | 66.48 1.47 9 - | 41.30 1.39 10 - | 25.16 1.24 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 28.1 3.8 10 0.91 | 8.529 ** 0.254 10 0.92 | 1.894 0.316 10 1.10 | 2.030 0.674 10 0.83 | 66.44 1.28 10 1.00 | 41.71 2.18 10 1.01 | 24.73 2.00 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 29.8 2.6 10 0.97 | 8.735 0.590 10 0.94 | 2.007 0.325 10 1.16 | 1.784 0.635 10 0.73 | 65.87 2.39 10 0.99 | 41.11 2.06 10 1.00 | 24.76 2.03 10 0.98 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.6 3.7 10 1.06 | 8.617 * 0.503 10 0.93 | 2.147 * 0.325 10 1.25 | 1.522 0.615 10 0.62 | 65.97 1.76 10 0.99 | 42.49 1.31 10 1.03 | 23.48 1.78 10 0.93 |
Sex: Male |
|
|
| Reporting Biochemistry |
|
| |
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G1] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.64 0.10 9 - | 2.592 0.033 9 - | 141.0 0.8 10 - | 5.09 0.30 10 - | 100.5 0.5 10 - | 1.747 0.191 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 1.70 0.19 10 1.03 | 2.620 0.064 10 1.01 | 142.0 1.2 10 1.01 | 5.12 0.30 10 1.01 | 101.7 * 0.8 10 1.01 | 2.034 ** 0.122 10 1.16 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.67 0.19 10 1.02 | 2.626 0.074 10 1.01 | 141.1 1.5 10 1.00 | 4.99 0.20 10 0.98 | 101.3 1.2 10 1.01 | 1.968 ** 0.179 10 1.13 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 1.83 0.17 10 1.11 | 2.657 0.064 10 1.02 | 141.6 2.0 10 1.00 | 4.95 0.18 10 0.97 | 99.9 1.6 10 0.99 | 2.065 ** 0.116 10 1.18 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
AST (U/L) [G] | ALT (U/L) [G] | ALP (U/L) [G] | GGT (U/L) [G1] | CK (U/L) [G] | TBIL (umol/L) [G1] | UREA (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 78.0 10.7 7 - | 55.9 14.4 7 - | 100.6 26.1 7 - | 1.5 0.0 7 - | 295.7 114.0 7 - | 1.25 0.00 7 - | 8.09 1.15 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 71.9 7.5 9 0.92 | 54.6 17.9 9 0.98 | 93.2 23.3 9 0.93 | 1.5 0.0 9 1.00 | 242.9 83.5 9 0.82 | 1.25 0.00 9 1.00 | 7.43 0.78 9 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 68.3 6.1 10 0.88 | 49.9 13.2 10 0.89 | 94.3 24.1 10 0.94 | 1.5 0.0 10 1.00 | 259.2 98.3 10 0.88 | 1.25 0.00 10 1.00 | 7.70 0.89 10 0.95 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 73.7 9.9 10 0.94 | 59.8 26.8 10 1.07 | 135.4 38.1 10 1.35 | 2.2 1.5 10 1.47 | 296.8 105.5 10 1.00 | 1.25 0.00 10 1.00 | 7.66 1.24 10 0.95 |
Sex: Female |
|
|
|
| Reporting Biochemistry |
|
|
|
CREAT (umol/L) [G] | GLUC (mmol/L) [G] | CHOL (mmol/L) [G] | TRIG (mmol/L) [G] | TPROT (g/L) [G] | ALB (g/L) [G] | GLOB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 34.0 3.5 7 - | 8.577 0.866 7 - | 1.589 0.267 7 - | 1.421 0.304 7 - | 65.96 2.49 7 - | 45.56 2.60 7 - | 20.40 2.41 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 34.8 4.5 9 1.02 | 7.801 0.566 9 0.91 | 1.512 0.297 9 0.95 | 1.021 0.261 9 0.72 | 67.76 2.46 9 1.03 | 46.70 1.56 9 1.03 | 21.06 1.80 9 1.03 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 32.7 4.0 10 0.96 | 8.161 0.464 10 0.95 | 1.647 0.235 10 1.04 | 1.268 0.529 10 0.89 | 68.21 2.48 10 1.03 | 46.77 2.62 10 1.03 | 21.44 2.18 10 1.05 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 32.4 4.5 10 0.95 | 8.082 0.391 10 0.94 | 1.913 * 0.234 10 1.20 | 0.978 0.268 10 0.69 | 67.82 2.61 10 1.03 | 46.97 2.19 10 1.03 | 20.85 1.87 10 1.02 |
Sex: Female |
|
| Reporting Biochemistry |
|
| ||
A/G (ratio) [G] | CA (mmol/L) [G] | NA (mmol/L) [G] | K (mmol/L) [G] | CL (mmol/L) [G] | PHOS (mmol/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 2.27 0.35 7 - | 2.574 0.035 7 - | 140.3 1.1 7 - | 4.43 0.42 7 - | 103.6 1.4 7 - | 1.227 0.104 7 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 2.24 0.21 9 0.99 | 2.581 0.060 9 1.00 | 140.2 1.2 9 1.00 | 4.78 0.29 9 1.08 | 102.9 1.2 9 0.99 | 1.436 0.206 9 1.17 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 2.18 0.30 10 0.96 | 2.591 0.053 10 1.01 | 140.6 0.8 10 1.00 | 4.45 0.20 10 1.00 | 104.1 1.3 10 1.01 | 1.379 0.205 10 1.12 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.30 0.28 10 1.01 | 2.664 ** 0.039 10 1.03 | 140.4 1.1 10 1.00 | 4.86 * 0.27 10 1.10 | 102.2 0.9 10 0.99 | 1.312 0.263 10 1.07 |
Table 15. Summary of Coagulation Values
F0 Animals – Males, Day: 137 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.55 0.20 10 - | 13.51 1.35 10 - | 2.278 0.177 10 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.43 0.15 10 0.99 | 12.52 1.72 10 0.93 | 2.282 0.122 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.30 ** 0.14 10 0.98 | 12.35 1.14 10 0.91 | 2.286 0.185 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.26 ** 0.12 10 0.97 | 11.77 1.51 10 0.87 | 2.454 * 0.114 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Table 16. Summary of Coagulation Values
F0 Animals – Females, Day: 115 Relative to Start Date
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.19 0.21 9 - | 11.17 1.22 9 - | 1.713 0.292 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.48 0.34 10 1.03 | 12.17 1.55 10 1.09 | 1.612 0.157 10 0.94 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.27 0.32 9 1.01 | 11.82 1.49 9 1.06 | 1.743 0.316 9 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.39 0.73 10 1.02 | 11.16 2.64 10 1.00 | 1.682 0.425 10 0.98 |
[G] - Anova & Dunnett
Table 17. Summary of Coagulation Values
F1 Animals - Cohort 1A, Day: 70 Relative to Start Date
Sex: Male |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.50 0.14 9 - | 12.27 1.80 9 - | 2.580 0.172 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.40 0.11 10 0.99 | 12.79 2.10 10 1.04 | 2.516 0.141 10 0.98 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.34 * 0.14 10 0.98 | 12.09 2.30 10 0.99 | 2.655 0.219 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.33 * 0.05 9 0.98 | 11.51 2.25 9 0.94 | 2.707 0.250 9 1.05 |
[G] - Anova & Dunnett: * = p ≤ 0.05
Sex: Female |
|
| Reporting Coagulation |
|
PT (sec) [G] | APTT (sec) [G] | FIB (g/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 10.31 0.26 9 - | 12.21 1.52 8 - | 2.116 0.251 9 . |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 10.17 0.17 9 0.99 | 12.33 1.57 9 1.01 | 2.128 0.172 9 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 10.15 0.14 10 0.98 | 11.65 0.99 10 0.95 | 2.186 0.194 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 10.02 ** 0.17 10 0.97 | 11.71 1.65 10 0.96 | 2.182 0.099 10 1.03 |
[G] - Anova & Dunnett: ** = p ≤ 0.01
Table 18. Summary of Hematology Values
F0 Animals – Males Day: 137 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G1] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 4.651 0.478 10 - | 0.961 0.257 10 - | 3.480 0.289 10 - | 0.065 0.014 10 - | 0.116 0.046 10 - | 0.008 0.004 10 - | 0.019 0.006 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 4.340 0.805 10 0.93 | 0.769 0.202 10 0.80 | 3.390 0.784 10 0.97 | 0.062 0.018 10 0.95 | 0.096 0.024 10 0.83 | 0.006 0.005 10 0.75 | 0.016 0.008 10 0.84 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.469 0.783 10 0.96 | 0.687 * 0.169 10 0.71 | 3.605 0.697 10 1.04 | 0.057 0.021 10 0.88 | 0.089 0.023 10 0.77 | 0.010 0.000 10 1.25 | 0.019 0.007 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.192 0.960 10 1.12 | 0.854 0.199 10 0.89 | 4.126 0.908 10 1.19 | 0.079 0.028 10 1.22 | 0.092 0.040 10 0.79 | 0.009 0.006 10 1.13 | 0.027 0.013 10 1.42 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 8.813 0.449 10 - | 153.6 4.7 10 . | 0.4464 0.0164 10 - | 50.68 1.37 10 - | 17.43 0.57 10 - | 343.9 7.0 10 - | 11.88 0.31 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.630 0.303 10 0.98 | 150.7 6.5 10 0.98 | 0.4441 0.0155 10 0.99 | 51.52 2.26 10 1.02 | 17.46 1.03 10 1.00 | 338.9 7.5 10 0.99 | 12.00 0.48 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.572 0.317 10 0.97 | 149.6 5.2 10 0.97 | 0.4388 0.0129 10 0.98 | 51.25 1.89 10 1.01 | 17.47 0.71 10 1.00 | 341.1 6.6 10 0.99 | 12.01 0.52 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.573 0.220 10 0.97 | 147.4 5.4 10 0.96 | 0.4364 0.0139 10 0.98 | 50.92 1.39 10 1.00 | 17.17 0.55 10 0.99 | 337.5 4.5 10 0.98 | 12.16 0.48 10 1.02 |
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 781.6 146.2 10 - | 155.83 19.62 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 765.3 76.5 10 0.98 | 142.51 41.33 10 0.91 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 760.4 92.6 10 0.97 | 160.37 18.17 10 1.03 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 883.4 160.3 10 1.13 | 171.47 21.32 10 1.10 |
Table 19. Summary of Hematology Values
F0 Animals – Females Day: 115 Relative to Start Date
Sex: Female |
| Reporting Hematology | ||||||
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.168 1.072 8 - | 1.669 0.559 8 - | 3.265 0.790 8 - | 0.109 0.031 8 - | 0.095 0.038 8 - | 0.011 0.006 8 - | 0.023 0.009 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.529 0.603 10 1.07 | 1.737 0.408 10 1.04 | 3.564 0.620 10 1.09 | 0.113 0.025 10 1.04 | 0.077 0.039 10 0.81 | 0.012 0.008 10 1.07 | 0.025 0.010 10 1.11 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 5.866 1.252 10 1.14 | 1.903 0.424 10 1.14 | 3.694 0.894 10 1.13 | 0.143 0.060 10 1.31 | 0.091 0.038 10 0.96 | 0.013 0.007 10 1.16 | 0.026 0.013 10 1.16 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.570 0.802 10 1.08 | 1.762 0.315 10 1.06 | 3.555 0.672 10 1.09 | 0.144 0.051 10 1.32 | 0.073 0.013 10 0.77 | 0.008 0.004 10 0.71 | 0.028 0.011 10 1.24 |
Sex: Female |
| Reporting Hematology | ||||||
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.958 0.449 8 - | 157.3 3.4 8 . | 0.4679 0.0124 8 - | 58.88 2.37 8 - | 19.80 0.92 8 - | 336.4 5.2 8 - | 12.00 1.06 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.678 0.380 10 0.96 | 153.3 5.2 10 0.97 | 0.4515 0.0208 10 0.97 | 58.86 1.56 10 1.00 | 19.99 0.77 10 1.01 | 339.8 8.3 10 1.01 | 11.84 0.79 10 0.99 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.466 0.606 9 0.94 | 152.7 5.1 10 0.97 | 0.4468 0.0194 9 0.95 | 59.99 2.69 9 1.02 | 20.48 1.21 9 1.03 | 341.6 7.1 9 1.02 | 11.82 0.97 10 0.99 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.528 0.566 10 0.95 | 151.3 8.8 10 0.96 | 0.4428 0.0316 10 0.95 | 58.85 1.83 10 1.00 | 20.10 1.07 10 1.02 | 341.5 10.8 10 1.02 | 12.17 1.05 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 894.6 119.2 8 - | 177.99 20.44 8 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 794.3 132.8 10 0.89 | 178.18 31.61 10 1.00 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 872.4 121.5 9 0.98 | 161.09 22.77 10 0.91 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 833.4 177.2 10 0.93 | 173.25 30.61 10 0.97 |
Table 20. Summary of Hematology Values
F1 Animals – Cohort 1A. Day 70 Relative to Start Date
Sex: Male |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G] | EOS (10^9/L) [G] | BASO (10^9/L) [G1] | LUC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 6.989 1.038 10 - | 0.930 0.183 10 - | 5.762 0.940 10 - | 0.121 0.043 10 - | 0.108 0.036 10 - | 0.008 0.009 10 - | 0.059 0.018 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.203 0.765 10 1.03 | 0.970 0.322 10 1.04 | 5.976 0.743 10 1.04 | 0.099 0.031 10 0.82 | 0.091 0.031 10 0.84 | 0.011 0.006 10 1.38 | 0.054 0.023 10 0.92 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.256 0.780 10 1.04 | 0.993 0.193 10 1.07 | 6.050 0.666 10 1.05 | 0.086 0.030 10 0.71 | 0.074 0.023 10 0.69 | 0.007 0.005 10 0.88 | 0.047 0.009 10 0.80 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.090 * 0.978 10 1.16 | 1.081 0.253 10 1.16 | 6.736 1.071 10 1.17 | 0.115 0.030 10 0.95 | 0.080 0.032 10 0.74 | 0.011 0.006 10 1.38 | 0.064 0.023 10 1.08 |
[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn
Sex: Male |
|
|
| Reporting Hematology |
|
|
| |
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 9.078 0.461 10 - | 163.2 4.9 10 . | 0.4747 0.0207 10 - | 52.33 1.52 10 - | 17.99 0.90 10 - | 343.7 10.9 10 - | 11.47 0.49 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 8.551 * 0.488 10 0.94 | 156.9 * 5.4 10 0.96 | 0.4471 ** 0.0181 10 0.94 | 52.33 2.09 10 1.00 | 18.37 0.87 10 1.02 | 350.8 6.2 10 1.02 | 11.59 0.52 10 1.01 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 8.522 ** 0.298 10 0.94 | 157.1 * 4.1 10 0.96 | 0.4519 * 0.0171 10 0.95 | 53.06 1.54 10 1.01 | 18.44 0.54 10 1.03 | 347.7 7.0 10 1.01 | 11.63 0.51 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 8.340 ** 0.285 10 0.92 | 154.7 ** 4.4 10 0.95 | 0.4456 ** 0.0166 10 0.94 | 53.45 1.51 10 1.02 | 18.56 0.67 10 1.03 | 347.4 7.6 10 1.01 | 11.84 0.32 10 1.03 |
[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
Sex: Male |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 763.2 131.0 10 - | 193.85 15.23 10 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 876.8 89.5 10 1.15 | 171.56 23.85 10 0.89 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 899.7 76.1 10 1.18 | 198.49 23.89 10 1.02 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 823.3 144.5 10 1.08 | 208.78 30.71 10 1.08 |
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
WBC (10^9/L) [G] | NEUT (10^9/L) [G1] | LYMPH (10^9/L) [G] | MONO (10^9/L) [G1] | EOS (10^9/L) [G] | BASO (10^9/L) [G] | LUC (10^9/L) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 5.104 1.013 9 - | 0.587 0.094 9 - | 4.321 0.997 9 - | 0.070 0.018 9 - | 0.081 0.019 9 - | 0.003 0.005 9 - | 0.038 0.010 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 5.207 1.111 9 1.02 | 0.671 0.142 9 1.14 | 4.371 1.058 9 1.01 | 0.053 0.017 9 0.76 | 0.074 0.028 9 0.92 | 0.004 0.005 9 1.33 | 0.029 0.014 9 0.76 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 4.656 0.870 10 0.91 | 0.708 0.238 10 1.21 | 3.780 0.791 10 0.87 | 0.060 0.021 10 0.86 | 0.067 0.020 10 0.83 | 0.003 0.005 10 0.90 | 0.034 0.008 10 0.90 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 5.515 1.387 10 1.08 | 0.681 0.323 10 1.16 | 4.640 1.417 10 1.07 | 0.076 0.039 10 1.09 | 0.079 0.031 10 0.97 | 0.004 0.005 10 1.20 | 0.037 0.014 10 0.98 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn
Sex: Female |
|
|
|
| Reporting Hematology |
|
|
|
RBC (10^12/L) [G] | HGB (g/L) [G] | HCT (L/L) [G] | MCV (fL) [G] | MCH (pg) [G] | MCHC (g/L) [G] | RDWG (%) [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 7.842 0.436 9 - | 151.6 5.3 9 . | 0.4248 0.0164 9 - | 54.23 1.82 9 - | 19.38 0.95 9 - | 356.8 8.1 9 - | 10.70 0.38 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 7.868 0.225 9 1.00 | 148.8 6.5 9 0.98 | 0.4244 0.0173 9 1.00 | 53.91 1.31 9 0.99 | 18.89 0.64 9 0.97 | 350.7 8.5 9 0.98 | 10.91 0.36 9 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 7.867 0.362 10 1.00 | 149.7 5.4 10 0.99 | 0.4244 0.0143 10 1.00 | 54.01 2.25 10 1.00 | 19.06 0.93 10 0.98 | 352.6 4.3 10 0.99 | 10.68 0.35 10 1.00 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 7.744 0.368 10 0.99 | 144.3 5.5 10 0.95 | 0.4116 0.0156 10 0.97 | 53.20 1.94 10 0.98 | 18.64 0.75 10 0.96 | 350.5 6.8 10 0.98 | 10.82 0.33 10 1.01 |
Sex: Female |
| Reporting Hematology | |
PLT (10^9/L) [G] | RETIC (10^9/L) [G1] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 833.0 141.8 9 - | 183.07 34.30 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 801.3 145.2 8 0.96 | 178.02 22.41 9 0.97 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 796.3 97.2 9 0.96 | 185.18 28.61 10 1.01 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 910.6 58.3 10 1.09 | 177.80 38.40 10 0.97 |
Table 21. Summary of Reproductive Performance
F0 Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 25 25 25 5.3 5.1 25 - 19 76.0 6 24.0 100.0 25/25 | 25 25 25 3.2 2.5 25 -39.4 24 96.0 1 4.0 100.0 25/25 | 25 25 25 2.7 2.6 25 -49.2 24 96.0 1 4.0 100.0 25/25 | 25 24 24 3.0 2.7 24 -43.2 23 95.8 1 4.2 100.0 24/24 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 22. Summary of Reproductive Performance
F1 Animals - Cohort 1B Females
Sex: Female Day(s) Relative to Pairing (Litter: A | ) | 0 mg/kg/day Group 1 | 100 mg/kg/day Group 2 | 300 mg/kg/day Group 3 | 900 mg/kg/day Group 4 |
Group Size - Females |
N+ve N+ve Mean SD N %Diff N+ve % N+ve % % ProA | 20 20 20 2.7 1.0 20 - 20 100.0 0 0.0 100.0 20/20 | 20 20 20 3.9 3.7 20 45.3 18 90.0 2 10.0 100.0 20/20 | 20 20 20 3.5 2.7 20 32.1 19 95.0 1 5.0 100.0 20/20 | 20 20 20 2.5 1.2 20 -7.5 20 100.0 0 0.0 100.0 20/20 |
Paired - Females | |||||
Mated Females | |||||
Pre-coital Interval (Days) | |||||
Confirmed Mating Days 1-7 | |||||
Confirmed Mating Days 8-14 | |||||
Female Mating Index |
Mated Females - Pregnant or Not Pregnant with confirmed mating date (by sperm or plug).
Table 23. Mating Performance and Fertility
F0 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 6(1)* | 4 | 8 | 7(1) |
2 | 2 | 5 | 6 | 5 |
3 | 6 | 8 | 7 | 3 |
4 | 5 | 7 | 3 | 8 |
No clear indication of mating | 6(1) | 1 | 1 | 1 |
Mean number of nights to positive mating sign | 2.5 | 2.8 | 2.2 | 2.5 |
Number passing one estrus | 0 | 0 | 0 | 0 |
Number of males paired | 25 | 25 | 25 | 24 |
Number of siring males | 24 | 25 | 25 | 23 |
Male Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Male Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Male Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
Female Mating Index | 0.96 | 1.00 | 1.00 | 1.00 |
Female Fertility Index | 1.00 | 1.00 | 1.00 | 0.96 |
Female Pregnancy Index | 0.96 | 1.00 | 1.00 | 0.96 |
*Includes Animal 1524 that had 2 implant sites (no live or dead pups)
Table 24. Mating Performance and Fertility
F1 Animals
Number of Nights to Positive Mating Sign | Group/Dose Level (mg/kg/day) | |||
1 (0) | 2 (100) | 3 (300) | 4 (900) | |
Number of Animals (Number of these not becoming pregnant) | ||||
1 | 3 | 5 | 2 | 7 |
2 | 5 | 2 | 4 | 2 |
3 | 9 | 5(2) | 8 | 6 |
4 | 2 | 5 | 4 | 5 |
5 | 1(1) | 1 | 0 | 0 |
6 | 0 | 0 | 1 | 0 |
No clear indication of mating | 0 | 2(2) | 1(1) | 0 |
Mean number of nights to positive mating sign | 2.7 | 2.7 | 2.9 | 2.5 |
Number passing one estrus | 0 | 1 | 1 | 0 |
Number of males paired | 20 | 20 | 20 | 20 |
Number of siring males | 19 | 16 | 19 | 20 |
Male Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Male Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Male Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Female Mating Index | 1.00 | 0.90 | 0.95 | 1.00 |
Female Fertility Index | 0.95 | 0.89 | 0.95 | 1.00 |
Female Pregnancy Index | 0.95 | 0.80 | 0.95 | 1.00 |
Table 25. Summary of Duration of Gestation and Overall Litter Performance
F0 Animals
F0 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 24 | 25 | 25 | 23 |
Duration of Gestation (Days) 21 |
1 |
1 |
0 |
0 |
22 | 17 | 17 | 23 | 20 |
23 | 0 | 6 | 1 | 2 |
No clear indication of mating | 6 | 1 | 1 | 1 |
Mean Duration | 21.9 | 22.2 | 22.0 | 22.1 |
Number of females producing a live litter | 23 | 25 | 25 | 23 |
Gestation index as % | 96 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.0 ± 3.4 | 12.5 ± 2.4 | 12.6 ± 1.9 | 12.9 ± 1.6 |
Mean total number of pups born per litter ± standard deviation | 11.0 ± 3.0 | 11.2 ± 2.3 | 11.0 ± 2.5 | 11.0 ± 3.3 |
Mean number of live pups per litter ± standard deviation: Lactation Day 1 |
11.0 ± 3.0 |
11.1 ± 2.2 |
10.9 ± 2.6 |
11.1 ± 2.6 |
Lactation Day 4 | 10.9 ± 3.1 | 11.2 ± 2.2 | 10.9 ± 2.6 | 11.0 ± 2.7 |
Lactation Day 4a | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 7 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8± 0.7 | 7.7 ± 0.9 |
Lactation Day 14 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Lactation Day 21 | 7.7 ± 1.1 | 7.9 ± 0.4 | 7.8 ± 0.7 | 7.7 ± 0.9 |
Number of males on Lactation Day 1 (%) | 120 (47) | 135 (49) | 145 (53) | 116 (47) |
Number of females on Lactation Day 1 (%) | 134 (53) | 143 (51) | 127 (47) | 129 (53) |
a After cull of pups on PND 4 to standardise the size of the litter
Table 26. Summary of Duration of Gestation and Overall Litter Performance
F1 Animals
F1 Generation | Group/Dose Level (mg/kg/day) |
| ||
1 | 2 | 3 | 4 | |
| (0) | (100) | (300) | (900) |
Number Pregnant | 19 | 16 | 19 | 20 |
Duration of Gestation (Days) 21 |
1 |
3 |
1 |
1 |
22 | 15 | 9 | 17 | 18 |
23 | 3 | 4 | 1 | 1 |
Mean Duration | 22.1 | 22.1 | 22.0 | 22.0 |
Number of females producing a live litter | 19 | 16 | 19 | 20 |
Gestation index as % | 100 | 100 | 100 | 100 |
Mean number of implant sites per pregnancy ± standard deviation | 12.6 ± 1.7 | 12.3 ± 2.3 | 13.0 ± 1.6 | 12.5 ± 2.3 |
Mean total number of pups born per litter ± standard deviation* | 11.6 ± 1.9 | 11.1 ± 2.8 | 10.5 ± 2.8 | 11.3 ± 2.2 |
Mean number of live pups per litter ± standard deviation*: Lactation Day 1 |
11.3 ± 2.2 |
11.1 ± 3.0 |
10.4 ± 2.7 |
11.1 ± 2.1 |
Lactation Day 4 | 11.3 ± 2.2 | 11.0 ± 2.9 | 10.3 ± 2.8 | 11.0 ± 2.1 |
Lactation Day 4a | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 7 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 14 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.9 ± 0.5 |
Lactation Day 21 | 7.9 ± 0.5 | 7.7 ± 0.9 | 7.6 ± 1.2 | 7.8 ± 0.5 |
Number of males on Lactation Day 1 (%) | 96 (47) | 96 (54) | 93 (49) | 100 (45) |
Number of females on Lactation Day 1 (%) | 108 (53) | 81 (46) | 96 (51) | 121 (55) |
a After cull of pups on PND 4 to standardise the size of the litter
* Excludes Animal 1628 that had a total litter loss
Table 27. Summary of Urinalysis Values
F0 Animals - Males
Day: 131 Relative to Start Date
| Sex: Male | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G1] | |||
| 0 mg/kg/day | Mean | 4.32 | 1.0152 | 7.5 |
| Group 1 | SD | 2.94 | 0.0089 | 0.94 |
| N | 10 | 10 - | 10 - | |
| - | ||||
| 100 mg/kg/day | Mean | 5.64 | 1.0135 | 8 |
| Group 2 | SD | 2.99 | 0.0052 | 0.62 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.31 | 1 | 1.07 | |
| 300 mg/kg/day | Mean | 4.22 | 1.0215 | 8.15 |
| Group 3 | SD | 2.11 | 0.0086 | 0.63 |
| N | 10 | 10 | 10 | |
| tCtrl | 0.98 | 1.01 | 1.09 | |
| 900 mg/kg/day | Mean | 5.25 | 1.0300 ** | 7.35 0.71 |
| Group 4 | SD | 2.39 | 0.0133 | 10 |
| N | 10 | 10 | 0.98 | |
| tCtrl | 1.22 | 1.01 |
Table 28. Summary of Urinalysis Values
F0 Animals - Females
Day: 108 Relative to Start Date
| Sex: Female | Reporting Urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G1] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 4.85 | 1.0188 | 8.6 |
| Group 1 | SD | 2.33 | 0.0052 | 0.32 |
| N | 10 - | 10 - | 10 - | |
| 100 mg/kg/day | Mean | 5.23 | 1.0192 | 8.45 |
| Group 2 | SD | 2.96 | 0.0069 | 0.16 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.08 | 1 | 0.98 | |
| 300 mg/kg/day | Mean | 6.01 | 1.0199 | 8.2 |
| Group 3 | SD | 1.68 | 0.0039 | 0.42 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.24 | 1 | 0.95 | |
| 900 mg/kg/day | Mean | 7.05 2.58 | 1.0218 0.0051 | 8.05 * |
| Group 4 | SD | 10 | 10 | 0.72 |
| N | 1.45 | 1 | 10 | |
| tCtrl | 0.94 |
[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05
Table 29. Summary of Urinalysis Values
F1 Animals - Cohort 1A
Day: 60 Relative to Start Date
| Sex: Male | Reporting urinalysis | |||
| VOLUME | SPECIFIC | URINE pH | ||
| (mL) | GRAVITY | [G] | ||
| [G] | [G] | |||
| 0 mg/kg/day | Mean | 2.67 | 1.0182 | 8.11 |
| Group 1 | SD | 1.87 | 0.0078 | 0.6 |
| N | 10 - | 10 - | 9 - | |
| 100 mg/kg/day | Mean | 3.07 | 1.0168 | 7.9 |
| Group 2 | SD | 0.95 | 0.0123 | 0.61 |
| N | 10 | 10 | 10 | |
| tCtrl | 1.15 | 1 | 0.97 | |
| 300 mg/kg/day | Mean | 1.96 | 1.0302 * | 7.11 ** 0.70 |
| Group 3 | SD | 1.25 | 0.0118 | 9 |
| N | 10 | 10 | 0.88 | |
| tCtrl | 0.73 | 1.01 | ||
| 900 mg/kg/day | Mean | 2.94 1.43 | 1.0346 ** | 6.25 ** |
| Group 4 | SD | 10 | 0.0091 | 0.42 |
| N | 1.1 | 10 | 10 | |
| tCtrl | 1.02 | 0.77 |
Day: 60 Relative to Start Date
Sex: Female |
|
| Reporting Urinalysi | s |
VOLUME (mL) [G] | SPECIFIC GRAVITY [G] | URINE pH [G] | ||
0 mg/kg/day
Group 1 | Mean SD N
| 1.88 1.25 10 - | 1.0226 0.0117 10 - | 7.00 0.94 9 - |
100 mg/kg/day
Group 2 | Mean SD N tCtrl | 3.20 2.88 10 1.70 | 1.0161 0.0086 10 0.99 | 7.15 0.91 10 1.02 |
300 mg/kg/day
Group 3 | Mean SD N tCtrl | 1.33 1.08 10 0.71 | 1.0333 0.0118 10 1.01 | 6.61 0.55 9 0.94 |
900 mg/kg/day
Group 4 | Mean SD N tCtrl | 2.61 1.55 9 1.39 | 1.0267 0.0087 9 1.00 | 6.44 0.68 9 0.92 |
[G] - Anova & Dunnett
Stability analysis The stability of the test substance suspensions over a period of 7 days at room temperature was demonstrated
Homogeneity analysis of the test substance preparations The homogeneous distribution of the test substance in the vehicle (1% CMC) was demonstrated
Concentration control analyses of the test substance preparations The results of the analysis of the aqueous test substance preparations confirmed the correctness of the prepared concentrations. The analytical values of the samples corresponded to the expected values within the limits of the analytical method, i.e. were always above 90% and below 110% of the nominal concentrations.
Tab. 1 Total soft tissue variations
| Test group 0, 0 mg/kg bw/d | Test group 1, 50 mg/kg bw/d | Test group 2, 150 mg/kg bw/d | Test group 3, 500 mg/kg bw/d | |||
| Litter Fetuses | N | 25 | 25 | 25 | 24 | |
| N | 118 | 119 | 112 | 105 | ||
| Fetal incidence | N% | 3 (2.5) | 2 (1.7) | 2 (1.8) | 10 (9.5) | |
| Litter incidence | N% | 3 (12) | 2 (8.0) | 2 (8.0) | 8 (33) | |
| Affected fetuses/litter | Mean % | 2.3 | 1.8 | 1.8 | 10.9* | |
mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent
* = p ≤ 0.05 (Wilcoxon-test [one-sided])
Tab. 2 Occurrence of statistically significantly increased fetal skeletal variations (expressed as mean percentage of affected fetuses/litter)
| Finding | Test group 0, 0 mg/kgbw/d | Test group 1, 50 mg/kgbw/d | Test group 2, 150 mg/kgbw/d | Test group 3, 500 mg/kgbw/d | HCDMean %(range) |
| Incomplete ossification ofnasal; unchanged cartilage | 2.1 | 2.1 | 5.2 | 9.9* | 1.4(0.0 – 10.1) |
| Dumbbell ossification ofthoracic centrum;dumbbell-shaped cartilageof centrum | 1.4 | 4.2 | 4.3 | 7.5* | 4.9(0.0 – 19.4) |
| Supernumerary thoracicvertebra | 3.3 | 2.3 | 3.5 | 13.3** | 3.9(0.8 – 8.0) |
| Misshapen sternebra;unchanged cartilage | 65.0 | 68.3 | 70.3 | 75.0* | 52.7(29.5 – 66.8) |
| Supernumerary rib (14th);cartilage present | 3.5 | 9.2* | 11.7* | 6.8 | 5.9(1.4 – 11.7) |
mg/kg bw/d = milligram per kilogram body weight per day; HCD = Historical control data; % = per cent
* = p ≤ 0.05 (Wilcoxon-test [one-sided])
** = p ≤ 0.01 (Wilcoxon-test [one-sided])
Tab. 3 Total unclassified cartilage observations
| Test group 0, 0 mg/kg bw/d | Test group 1, 50 mg/kg bw/d | Test group 2, 150 mg/kg bw/d | Test group 3, 500 mg/kg bw/d | ||
| Litter | N | 25 | 25 | 25 | 24 |
| Fetuses | N | 130 | 129 | 128 | 121 |
| Fetal incidence | N (%) | 103 (79) | 97 (75) | 107 (84) | 107 (88) |
| Litter incidence | N (%) | 24 (96) | 25 (100) | 25 (100) | 23 (96) |
| Affectedfetuses/litter | Mean | 79.3 | 76.2 | 85.1 | 87.1* |
mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent
* = p ≤ 0.05 (Wilcoxon-test [one-sided])
Tab. 4 Total fetal variations
| Test group 0, 0 mg/kg bw/d | Test group 1, 50 mg/kg bw/d | Test group 2, 150 mg/kg bw/d | Test group 3, 500 mg/kg bw/d | ||
| Litter | N | 25 | 25 | 25 | 24 |
| Fetuses | N | 248 | 248 | 240 | 226 |
| Fetal incidence | N (%) | 133 (54) | 130 (52) | 130 (54) | 131 (58) |
| Litter incidence | N (%) | 25 (100) | 25 (100) | 25 (100) | 24 (100) |
| Affectedfetuses/litter | Mean | 53.4 | 52.6 | 54.8 | 58.5* |
mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent
* = p ≤ 0.05 (Wilcoxon-test [one-sided])
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 150 mg/kg bw/day
- Species:
- rabbit
- Quality of whole database:
- There are two studies avaiilable for consideration, both were conducted according to OECD guideline 414 and to GLP, and both Klimisch code 1. One was conducted in the rat and one in the rabbit. The overall quality of the database is therefore considered to be high. The most senstive species is concluded to be the rabbit and the NOAEL observed in this study is the taken as the overall result.
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Additional information
OECD Guideline 414 with rats:
range finding study
In the course of a range finding study the test item was administered once daily by oral gavage to groups of 10 mated Wistar rats at dose levels of 0 (vehicle control; test group 0), 150 mg/kg bw/d (test group 1) and 400 mg/kg bw (test group 2) from gestation days (GD) 6 to 19. Effects of beginning systemic toxicity were limited to the high dose group, i.e. salivation in 6/10 animals, increased relative liver (110%) and kidney weights (108%) as well as increased alaninaminotransferase. Dose levels 50, 150 and 500 mg/kg bw/day were regarded to be appropriate to test the material in an OECD guideline 414 setup for its potential effects on rat offspring without causing exaggerated toxicity in their mothers.
main study OECD 414
The test substance was tested for its prenatal developmental toxicity in Wistar rats. The test substance was administered as an aqueous suspension to groups of 25 time-mated female Wistar rats by gavage at doses of 50, 150, and 500 mg/kg body weight/day (mg/kg bw/d) on gestation days (GD) 6 through 19. The control group, consisting of 25 females, was dosed with the vehicle (1% Carboxymethylcellulose suspension in drinking water (1% CMC)) in parallel. A standard dose volume of 10 mL/kg body weight was used for each test group. At terminal sacrifice on GD 20, 24-25 females per group had implantation sites.
Food consumption and body weights of the animals were recorded regularly throughout the study period. The state of health of the animals was checked each day. On GD 20, all females were sacrificed by cervical dislocation (under isoflurane anesthesia) and assessed by gross pathology (including weight determinations of the unopened uterus and the placentae). For each dam, corpora lutea were counted and number and distribution of implantation sites (differentiated between resorptions, live and dead fetuses) were determined. The fetuses were removed from the uterus, sexed, weighed and further investigated for external findings. Thereafter, one half of the fetuses of each litter were examined for soft tissue findings and the remaining fetuses for skeletal (inclusive cartilage) findings.
The stability of the test substance preparations was demonstrated over a period of 7 days at room temperature. The homogeneous distribution of the test substance in the vehicle (1% CMC suspension in drinking water) was confirmed. The correctness of the prepared concentrations was shown.
Under the conditions of this prenatal developmental toxicity study, the oral administration to pregnant Wistar rats from implantation to one day prior to the expected day of parturition (GD 6-19) at a dose of 500 mg/kg bw/d caused no evidence of maternal toxicity.
In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 500 mg/kg bw/d.
The no observed adverse effect level (NOAEL) for prenatal developmental toxicity is 500 mg/kg bw/d. There were no toxicologically relevant adverse fetal findings evident. Slightly higher rates of individual non-specific fetal variations at a dose of 500 mg/kg bw/d were considered to be an effect of the treatment, but not as adverse. Hence the no observed effect level (NOEL) for prenatal developmental toxicity is 150 mg/kg bw/d.
Toxicity to reproduction: other studies
Description of key information
OECD 414 with Rabbits
A range finding study was conducted and based on the results of thsi study and at the request of the sponsor three dose levels were selected for the main OECD 414 study.
The test substance was administered as an aqueous suspension to groups of 25 inseminated female New Zealand White rabbits orally by gavage in doses of 15, 50 and 150 mg/kg body
weight/day (mg/kg bw/d) on gestation days (GD) 6 through 28. The vehicle control group, consisting of 25 females, was dosed with the vehicle 0.5% Carboxymethylcellulose suspension in
drinking water (0.5% CMC) in parallel. A standard dose volume of 10 mL/kg body weight was used for each test group
Oral admisnistration of the test substance to pregnant New Zealand White rabbits from implantation to one day prior to the expected day of parturition (GD 6-28) caused evidence of maternal toxicity at the highdose (150 mg/kg bw/d), such as adverse clinical findings. In conclusion, the no observed adverse effect level (NOAEL) for maternal toxicity is 50 mg/kg bw/d.
Since there was no evidence for toxicologically relevant adverse effects of the test substance on fetal morphology at any dose, the no observed adverse effect level (NOAEL)
for prenatal developmental toxicity is the highest dose of 150 mg/kg bw/d.
Justification for classification or non-classification
The test material is not classified as a reproductive toxicant.
Reproductive toxicity includes adverse effects on sexual function and fertility as well as developmental toxicity in the offspring.
Weight of evidence from all three study results indicate that there are no relevant adverse effects on either fertility or relelvent fetal findings i.e fertility and developmental toxicty even at the highest doses so there is no evidence that the test material is a reproductive toxicant and so classification is not required.
Additional information
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