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Registration Dossier
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EC number: 231-272-0 | CAS number: 7473-98-5
Toxicity to reproduction
Administrative data
- 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
Cross-reference
- Reason / purpose for cross-reference:
- read-across source
- Remarks:
- Hydroxycyclohexyl phenyl ketone
Reference
- 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.
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
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 022
Materials and methods
Test guidelineopen allclose all
- 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.
Test material
- Reference substance name:
- Hydroxycyclohexyl phenyl ketone
- EC Number:
- 213-426-9
- EC Name:
- Hydroxycyclohexyl phenyl ketone
- Cas Number:
- 947-19-3
- Molecular formula:
- C13H16O2
- IUPAC Name:
- (1-hydroxycyclohexyl)-phenylmethanone
- Test material form:
- solid: particulate/powder
- Details on test material:
- Purity 99.7-99.9%
Constituent 1
- 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
Test animals
- 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
Administration / exposure
- 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)
Doses / concentrationsopen allclose all
- 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
Examinations
- 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
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- 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 / performance (P0)
- 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.
Effect levels (P0)
- 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
Target system / organ toxicity (P0)
- Critical effects observed:
- no
Results: F1 generation
General toxicity (F1)
- 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.
Developmental neurotoxicity (F1)
- 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 (F1)
- 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.
Effect levels (F1)
- 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
Target system / organ toxicity (F1)
- Critical effects observed:
- no
Results: F2 generation
General toxicity (F2)
- 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
Developmental neurotoxicity (F2)
- Behaviour (functional findings):
- not examined
Developmental immunotoxicity (F2)
- 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.
Effect levels (F2)
- 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
Target system / organ toxicity (F2)
- Critical effects observed:
- no
Overall reproductive toxicity
- Key result
- Reproductive effects observed:
- no
- Lowest effective dose / conc.:
- 900 mg/kg bw/day
- Treatment related:
- no
Any other information on results incl. tables
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
Applicant's summary and conclusion
- 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.
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