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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 October 2015 - 09 May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Test material form:
liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL- Source and lot/batch No.of test material: JBLE0012T- Expiration date of the lot/batch: May 2016STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: Controlled room temperature protected from lightTREATMENT OF TEST MATERIAL PRIOR TO TESTING- Treatment of test material prior to testing: None

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS - Source: Charles River Laboratories, Sulzfeld, Germany - Age at study initiation: 11 weeks old at mating - Weight at study initiation: 197-233 g - Fasting period before study: - Housing: - Diet (e.g. ad libitum): ssniff SM R/M "Autoclavable Complete Feed for Rats and Mice - Breeding and Maintenance" - Water (e.g. ad libitum): tap water - Acclimation period: 5 days ENVIRONMENTAL CONDITIONS - Temperature (°C): 20.0 - 23.8°C - Humidity (%): 39-70% - Air changes (per hr): 15-20/hour - Photoperiod (hrs dark / hrs light):12 / 12 IN-LIFE DATES: From: 17 November 2015 To: 07 December 2015

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: VEHICLE - Justification for use and choice of vehicle (if other than water): corn oil - Lot/batch no. (if required): MKBQ9948V / MKBS6944V- 3 mL/kg
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis of test substance for concentration and/or homogeneity by validated GC method.Duplicate samples (top, middle and bottom samples) taken from the test formulation two times during the study (second and last weeks of treatment). One duplicate sample taken on each occasion from vehicle control for concentration measurement.
Details on mating procedure:
- Impregnation procedure: cohoused - If cohoused: - M/F ratio per cage: 1:1 - Length of cohabitation: 2 hours - Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
GD 6 to GD 19
Frequency of treatment:
Daily
Duration of test:
Until GD 20
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
24 mated females/group
Control animals:
yes
Details on study design:
- Dose selection rationale: based on available data, including an OECD 422 study

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes - Time schedule: Twice daily each day DETAILED CLINICAL OBSERVATIONS: Yes / No / No data - Time schedule: at the onset of treatment (GD 6) then weekly BODY WEIGHT: Yes - Time schedule for examinations: GD 0, 3, 6, 8, 10, 12, 14, 16, 18 and 20 FOOD CONSUMPTION: Yes - Time schedule: GD 0, 3, 6, 8, 10, 12, 14, 16, 18 and 20 POST-MORTEM EXAMINATIONS: Yes - Sacrifice on gestation day 20 - Organs examined: viscera, ovaries, uterus, OTHER:
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes Examinations included: - Gravid uterus weight: Yes - Number of corpora lutea: Yes - Number of implantations: Yes - Number of early resorptions: Yes - Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes: all per litter - Soft tissue examinations: Yes: half per litter - Skeletal examinations: Yes: half per litter - Head examinations: Yes:half per litter
Statistics:
SPSS PC+4.0
Historical control data:
Yes

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, non-treatment-related
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Statistically significantly decreased bodyweight and bodyweight gain were seen in the high dose group on several days/periods. However, the difference in terminal bodyweight and bodyweight gain during the study (GD 0 - 20) was less than 10%. Furthermore, the bodyweight loss duing the treatment period (GD 6 - 20) was not considered statistically significant. Therefore, these findings were not considered as clear treatment-related adverse effects. A similar but stronger effect was detected for the corrected terminal bodyweight, corrected bodyweight gain and corrected net bodyweight gain. The corrected bodyweight gain and corrected net bodyweight gain values were significantly lower compared to the control group (by 22 and 29%, respectively). These adverse effects were considered related to administration of the test substance.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
A slight effect on food consumption was observed for all test groups. Statistically significant decreased food consumption was seen for the high dose group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Maternal developmental toxicity

Number of abortions:
not examined
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 examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
not examined
Other effects:
not examined

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
not examined
Changes in litter size and weights:
not examined
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not examined

Effect levels (fetuses)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Remarks on result:
other: Embryotoxicity
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Remarks on result:
other: Foetotoxicity
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Remarks on result:
other: Teratogenicity

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Applicant's summary and conclusion

Executive summary:

A study was conducted to assess the effect of amine synergist on the embryonic and fetal development of Hannover Wistar rats in their first pregnancy according to OECD Guideline 414, in compliance with GLP. Groups of 24 pregnant females were administered the substance diluted in corn oil (3 mL/kg) at 0, 100, 300 and 1000 mg/kg/day from Gestation Day 6 to 20. Cesarian sections, necrospy of dams and examinations of uterine content were performed on GD 20.

Dose formulations were analysed for test substance concentration twice during the treatment period using a validated GC method. Parameters monitored during the study included mortality and clinical observations, body weight, body weight gain and individual food consumtion. Maternal reproductive parameters associated with uterine examination were evaluated, and the foetuses were weighed ande xamined for external, visceral and skeletal abnormalities. Placentas were examined macroscopically. The number of confirmed pregnant, evaluated dams in the dose groups treated was 24 in the control group and 23, 24 and 24 in the low, mid and high dose groups, respectively.

All formulations were within the range of 92 to 107% of nominal concentration an were found to be homogenous. No test item was detected in the control samples. Based on these results, test item formulations were considered suitable for the study purposes. There was no unscheduled mortality or treatment related clinical signs in the study. Statistically significantly decreased body weight or body weight gain values were observed for the High dose group (100 mg/kg bw/day) on several days / periods. However, the difference in the terminal body weight and the body weight gain during the study (GD0-20) was less than 10%. Furthermore the body weight loss during the treatment period GD6-20) was not considered statistically significant. Therefore these findings were not considered as a clear test item related adverse effect. A similar but stronger effect was detected for the corrected terminal body weight, corrected body weight gain and corrected net body weight gain. The corrected body weight gain and corrected net body weight gain values were significantly lower compared to the control group (by 22% and 29% respectively). These adverse effects were considered related to the administration of the test item. A slight effect on the food consumtpion was observed in the study for all test groups. Additionally, statistically significant decreased food consumption was observed for the high dose group. There were no toxicologically significant differences, or test item related-changes in the reproductive parameters examined in the conditions of this study. The number of corpora lutea, implantation sites and pre-implantation loss mean values were comparable to the control for the test item treated groups at all dose levels. There were no effects on the early and late embryonic loss, post-implantation (total resorption, including the early and late embryonic loss) or total intrauterine mortality in the test item-related dams. No remarkable internal or external observations were recorded for any pregnant animals during necropsy. No abnormalities were observed on the placentas in any examined groups. The mean number of viable foetuses in all test item treated groups as well as their sex distribution were comparable with the control.

The weight of foetuses in the low, mid and high dose groups did not differ significantly from the control mean, when evaluated by both litter mean and group mean.

The number of body weight retarded foetuses in the test item treated groups was lower than in the control group. All abnormalities observed at external, visceral or skeletal examination in this study were considered as incidental findings, which were also observed in the concurrent study control group. The findinds lauid within historical control (HC) data or were considered to be spontaneous events unrelated to treatment.

In conclusion, when administered daily by oral gavage to pregnant Hannover Wistar rats from gestation days GD 6 to 19, amine synergist caused a slight maternal body weight and food intake effect in the high dose group, demonstrating an adequate but relatively minor maternal toxicity. There was no evidence of any adverse maternal effects in the mid and low dose groups. There were no toxicologically significant differences, or test item related-changes in the reproductive parameters examined in the conditions of this study. No foetal effects were seen at external, visceral and/or skeletal examination of foetuses in the study which could be related to the test item administration. The following NOAELs were derived (Hargitai, 2016):

NOAELmaternal toxicity:300 mg/kg bw/day
NOAEL
embryotoxicity:1000 mg/kg bw/day
NOAEL foetotoxicity
: 1000 mg/kg bw/day
NOAELteratogenicity
: 1000 mg/kg bw/day