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Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data from J-check

Data source

Reference
Reference Type:
other: authoritative database
Title:
Repeated oral administration toxicity / reproductive developmental toxicity combined test using test material in rat
Author:
J-check
Year:
2010
Bibliographic source:
J-check, Environment Agency, Ministry of Health, Labour and Welfare,Ministry of Economy, Trade and Industry, and Ministry of the Environment, 2010

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Repeated oral administration toxicity / reproductive developmental toxicity combined test using test material in rat.
GLP compliance:
not specified
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Phthalimide
EC Number:
201-603-3
EC Name:
Phthalimide
Cas Number:
85-41-6
Molecular formula:
C8H5NO2
IUPAC Name:
1H-isoindole-1,3(2H)-dione
Details on test material:
- Name of test material (as cited in study report):1H-isoindole-1,3(2H)-dione
- Molecular formula :C8H5NO2
- Molecular weight :147.133g/mol
- Substance type: Organic

Test animals

Species:
rat
Strain:
other: Crj: CD (SD) IGS
Details on test animals or test system and environmental conditions:
Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Japan Co., Ltd.
- Age at study initiation: 10week old
- Weight at study initiation: 371.7 g (350 to 402 g) for males
231.5 g (195 to 257 g) for female
- Fasting period before study:
- Housing: bracket type metal wire net floor cage.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): CRF-1, Oriental Yeast Industry Co., Ltd, ad libitum
- Water (e.g. ad libitum): drinking water using tap water (Sapporo city) ad libitum
- Acclimation period:2 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 to 24 ° C
- Humidity (%):45 to 61%,
- Air changes (per hr): 10 to 15 times / hour
- Photoperiod (hrs dark / hrs light): 12 hours (lit from 8 am to 8 pm)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 1 w / v% aqueous solution of sodium carmellose
Details on exposure:
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material suspended in a 1 w / v% aqueous solution of sodium carmellose
DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): 1 w / v% aqueous solution of sodium carmellose
- Concentration in vehicle: 0,250,500,1000 mg/kg bw/day
- Amount of vehicle (if gavage): 10ml/kg bw
- Lot/batch no. (if required):
- Purity:
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:the day on which sperm was
confirmed in female vaginal plaque was taken as the 0th gestation
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.:No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]No data available
- After successful mating each pregnant female was caged (how):No data available

Duration of treatment / exposure:
Males:46
Females: 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation.
Frequency of treatment:
Daily
Duration of test:
Approx 54 days
Doses / concentrations
Remarks:
0,250,500,1000 mg/kg bw/day
No. of animals per sex per dose:
Total:96
0mg/kg bw/day:12 male and 12 female
250 mg/kg bw/day:12 male and 12 female
500 mg/kg bw/day:12 male and 12 female
1000mg/kg bw/day:12 male and 12 female
Control animals:
yes, concurrent vehicle

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period.
Parental animals observation and examinations
DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: On 1 day of administration (before administration), 2, 5, 7, 10 and 14 days of administration, after every 7 days (including the administration finish date) for males, for females 0, 1, , 7, 10, 14, 17, and 20, at 0, 1 and 4 days of nursing, and during the mating period

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Feed intake was measured for males except for the mating period, final administration day and autopsy date, and
for females on the same day as the body weight measurement day except for gestation day 0 and nursing 0 day,
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data
- Time schedule for examinations:
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes:
- Skeletal examinations: Yes:
- Head examinations: No data
Statistics:
Multiple sample χ ^ 2- test was performed on findings that showed one level of positive grade among sexual cycle, mating rate, conception rate, childbirth rate and nursing rate, and histopathological examination results, and in case of significant 2 Sample c 2 - Assay was performed. In addition, Fisher's direct probability test method was used when these tests failed. For other observations and findings showing positive grade of 2 or more out of the results of histopathological examination, after equality dispersal test of Bartlett, analyzed by one way analysis of variance or Kruskal-Wallis method, significant , The comparison group and the administration group were compared by Dunnett's test method or Mann-Whitney U-test method. For the test with the control group, the significance level was set at 5%.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups.
In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.

Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation.
In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation.
In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
no effects observed
Description (incidence and severity):
There was no significant difference in any of the test items compared with the control group in each group of male
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In males, no significant difference was observed in each treated group and control group.
In the females, the high weight of the body weight ratio of the lung was in the 250 mg / kg group and the high value of the weight and weight ratio of the spleen was observed in the 500 mg / kg group, but in the 1000 mg /kg group compared with the control group There was no significant difference.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Histopathological examination revealed eosinophilic corpuscle of the proximal tubular epithelium in the kidney in males in 1 to 3 cases in the 250, 500 and 1000 mg / kg groups, but in comparison with the control group No significant difference was observed and it was not considered to be an effect of test material administration no dose-dependent increase was observed in the collection of lung foam cells and the number of occurrences of lymphocyte infiltration of the prostate, and the other findings were also changed only in one case, which is not related to test material administration. Histological examination of females showed only one case in the 1000 mg / kg group, but peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and atrophy of the thymus were observed in the kidney
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Maternal developmental toxicity

Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
effects observed, treatment-related
Description (incidence and severity):
There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
changes in number of pregnant
clinical signs
early or late resorptions
food consumption and compound intake
gross pathology
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
pre and post implantation loss
total litter losses by resorption
urinalysis
Remarks on result:
other: No toxic effects obaserved

Maternal abnormalities

Abnormalities:
not specified
Localisation:
not specified

Results (fetuses)

Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Compared with the control group, the low value of body weight of 4 days of nursing in males of the 500 mg / kg group, the 4 days of nursing of body weight gain, the weight gain and the body weight gain rate were low in females as compared with the control group. Even in the 1000 mg / kg group, low values of weight of body
weight of 4 days of nursing were observed in male, 4 days of body weight in nursing and low value of body weight gain were observed in females.

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
no effects observed
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Deficits in the tail tips of the control group observed in general condition observation and one in each of the 1000 mg / kg group, and abdominal or cervical trauma in 1 case of each sex of 1000 mg / kg group were observed , No other abnormalities were found in any of the dead cases and autopsy of newborn sacrificed on the
4th day of nursing in any animals.
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
not specified

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
fetal/pup body weight changes
changes in postnatal survival
external malformations
Remarks on result:
other: No developmental toxic effects were observed

Fetal abnormalities

Abnormalities:
not specified
Localisation:
other: not specified

Overall developmental toxicity

Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

Influence of test material on reproductive performances of rats in combined repeat dose and reproductive/ developmental toxicity screening test

 

Item

0mg/kg

250mg/kg

500mg/kg

1000mg/kg

No of animals examined

12

12

12

12

No of pairs mated

12

12

12

12

No of pairs with successful copulation

12

12

12

12

Duration of mating (days. Mean ±SD.)

3.3± 1.1

2.9 ±1.2

3.7± 3.4

3.6 ±2.5

Copulation index(%)

100.0

100.0

100.0

100.0

No of pregnant animals

12

12

12

10

Fertility index(%)

100.0

100.0

100.0

83.3

a:(No. of pairs with successful copulation/ no. of pairs mated)x103

b:(No. of pregnant animals/ no. of pairs with successful copulation)x100

 

Influence of test material on developmental performances of rats in combined repeat dose and reproductive/ developmental toxicity screening test

Item

0 mg/kg

250mg/kg

500 mg/kg

1000 mg/kg

No of pregnant animals

12

12

12

12

No of corpora lutea

16.9± 2.4

17.5± 2.8

17.7 ±2.1(11)

18.1± 1.9

No of implantation sites

15.7 ±1.7

16.6 ±1.7

15.7± 4.2

15.5± 3.2

Implantation index‘(%)

93.3± 8.3

95.6 ±7.3

93.7 ±10.5

86.0 ±18.5

No of pups born

14.3± 1.7

15.6± 2.1

14.3 ±5.2

15.2± 3.2

Delivery index"(%)

91.2± 7.2

93.9 ±7.0

85.5± 27.3

98.2± 3.0**

Live pups born

 

 

 

 

No.

14.3± 1.7

15.4 ±2.1

15.5 ±2.6(11)

15.1 ±3.1

Live birth index‘(%)

100.0± 0.0

98.9± 2.5

99.5± 1.6(11)

99.4 ±1.8

Sex ratio (M/F)

1.21 ±0.63

1.36± 0.64

1.10 ±0.82

1.23 ±0.57

Dead pups born

 

 

 

 

No.

0.0 ±0.0

0.2± 0.4

0.1± 0.3(11)

0.1± 0.3

Gestation length(day)

22.5± 0.5

22.7± 0.5

22.7 ±0.2(11)

22.3 ±0.5

Gestation index‘(%)

100.0

100.0

91.7

100.0

Nursing index°(%)

100.0

100.0

100.0

100.0

Live pups on day 4

 

 

 

 

No

14.1 ±1.4

15.3 ±2.1

15.0 ±2.4(11)

14.1± 3.4

Viability index‘(%)

99.0 ±2.3

99.5 ±1.8

97.3 ±5.0(11)

91.1± 14.7

Body weight of pups(g)

 

 

 

 

Male Day 0

6.93 ±0.40

6.82 ±0.69

6.71 ±0.50(11)

6.41 ±0.62

Day 1

7.61 ±0.55

7.53± 0.91

7.24± 0.60(11)

6.94 ±0.94

Day 4

10.88± 0.86

10.62± 1.55

10.04 ±0.83*(11)

9.68 ±1.93*

Day 04 gain(g)

3.96 ±0.51

3.80 ±1.04

3.33 ±0.64(11)

3.27± 1.54

Body weight gain%

57.04± 5.54

55.48± 11.51

49.81± 10.29(11)

50.441 ±22.95

Female Day 0

6.58 ±0.40

6.44± 0.64

6.27 ±0.49(11)

6.09 ±0.56

Day 1

7.21± 0.47

7.06± 0.47

6.74± 0.59(11)

6.58± 0.81

Day 4

10.43 ±0.79

10.08 ±1.46

9.39 ±0.65**(11)

9.12± 1.71*

Day 0-4. gain(g)

3.85 ±0.47

3.63± 1.04

3.12± 0.46**(11)

3.03 ±1.42*

Body weight gain(%)

58.50 ±5.70

56.26 ±13.16

50.03 ±8.93*(11)

49.47± 22.16

 

Values are expressed as Mean ±S.D.

Values in parent theses are no. of animals examined.

Significantly different from 0 mg/kg group*p<0.05,**p <0.01

a:(No. of implantation sites/no. of corpora lutea)X100

b:(No of pups born/no. of implantation sites)X100

c:(No. or live pups born/ no. or pups born)X100

d:(No of females with live pups delivered/no. of pregnant females)X100

e:(No. of females nursing live pup/no. of females with normal delivery)X100

f: (No. of live pups on day 4/ no. of live pups born)X100

g:(Body weight gain/body weight on day 0)X100

Applicant's summary and conclusion

Conclusions:
The no effect level (NOEL) for reproduction of parent animals by repetitive oral administration of test material in this study was considered to be 1,000 mg / kg / day in male, 250 mg / kg / day for females, and 250 mg / kg / day for no effect on neonatal development (NOEL).
Executive summary:

The reproductive and developmental toxicity of test material was performed on male and female rats. The test material suspended in 1 w / v% aqueous solution of sodium carmellose in dose concentration0, 250, 500, 1000 mg/kg bw/day and administered orally by gavage. The dose concentration were selected on the bases on dose finding study (5, 70 and 1000 mg / kg).Exposure to test material for the males, 46 days including the mating period and for the females for 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation. For males and females on 14th day of administration, they were allowed to live together in the same group within 1 to 1 (random combination) from the evening for only 14 days and the day on which sperm was confirmed in female vaginal plaque was taken as the 0th gestation. For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period. For all female mated females, from the 21st day of pregnancy to the end of calving, the status of parturition, nursing behaviour, the number of total births, the number of surviving children and the number of dead children, the sex and the outer table of the infant were observed.

In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups. In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation. In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.

In reproductive tests, male and female mating ratios, female sexual cycles and conception rates, test material administration in the necropsy, weight and histopathological examination of reproductive organs (testis, epididymis and ovaries) and endocrine organs (pituitary, adrenal) No effect was observed. On the other hand, pathological examination of the infertile reproductive organs of 2 cases observed in the 1000 mg / kg group showed no abnormality and no influence by test material administration was observed.

There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.Three cases were unknown, and the weight of surviving newborn infants was decreasing. In this mother animals, body weight loss and low food intake were also observed during the nursing period, and histopathological examination revealed peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and thymic Atrophy was observed. Therefore, although only one case appeared, the possibility that test material administration might have affected the mother's ability to nurture was considered.

In addition, in one case of 500 mg / kg group where delivery was not observed until 26th day of pregnancy, laparotomy of left and right uterus and cervix was observed at necropsy. In the same example, five implantation marks were seen by observation with the naked eye, both of which are considered to be early embryonic deaths, and the relation with test material administration was observed. survival were observed, in neonates weighing transition observed low values of body weight nursing 4 days in male and female 500 and 1000 mg / kg group, In addition, in the 500 mg / kg females, the body weight gain and the body weight gain rate were low, and even in females of the 1000 mg / kg group, the body weight gain was low. But neonatal weight during delivery was not changed in each group treated with test material. Therefore, a decrease or milk migration nursing ability of dams by test material administration was believed to have affected the newborn weighing 500 and 1000 mg / kg group, including one example of 1000 mg / kg group described above, The mechanism could not be clarifiedunder this test condition. Hence The no effect level (NOEL) for reproduction of parent animals by repetitive oral administration of test material in this study was considered to be 1,000 mg / kg / day in male, 250 mg / kg / day for females, and 250 mg / kg / day for no effect on neonatal development (NOEL).