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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

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
Type:
Constituent
Test material form:
liquid: viscous

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Hannover Wistar rats (CRL:WI(Han)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Sulzfeld, Germany
- Age at study initiation: at least 13 weeks old at mating
- Weight at study initiation: 203 - 257 g
- Fasting period before study: no
- Housing: individual housing
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: at least 15 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.3 - 24.8°C
- Humidity (%): 33 - 65%
- Air changes (per hr): 15 - 20 per hour
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on exposure:
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 20, 60, 200 mg/ml
- Amount of vehicle (if gavage): 5 mg/kg
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test substance was dispersed in propyleneglycol. This dispersion was diluted with tetrahydrofuran (1:2) and directly measured by near infrared (NIR) spectroscopy between 908 and 1676 nm (transmission).
Details on mating procedure:
- If cohoused:
- M/F ratio per cage: 1 : 1
- Length of cohabitation: females with appropriate oestrous cycle were paired in the morning for approx. 2 hours
- Proof of pregnancy: vaginal plug or sperm in vaginal smear was referred to as day 0 of pregnancy
Duration of treatment / exposure:
from gestation day 6 till gestation day 19
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
24
Control animals:
yes, concurrent vehicle

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at the first day of treatment and then weekly

BODY WEIGHT: Yes
- Time schedule for examinations: on GD 0, 3, 6, 8, 10, 12, 14, 16, 18, 20

FOOD CONSUMPTION
- Food consumption for each animal determined on GD 0, 3, 6, 8, 10, 12, 14, 16, 18, 20
- mean daily diet consumption calculated as g food/kg body weight/day: Yes, for intervals of days on which body weight and food consumption were determined

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: the dams' viscera were examined macroscopically
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:
The statistical evaluation of data was performed with the program package SAS v9.2 in case of Provantis v.9, or SPSS PC+4.0 (SPSS Hungary Kft, Budapest) in case of data tabulated in Excel, by an appropriate statistical method.

In case of the SAS v9.2 software package (within the validated Provantis system) the following decision tree was applied automatically for statistical evaluation of numeric data. The normality and heterogeneity of variance between groups were checked by Shapiro-Wilk and Levene tests using the most appropriate data format (logtransformed when justified). Where both tests showed no significant heterogeneity, an Anova / Ancova (one-way analysis of variance) test was carried out. If the obtained result was positive, Dunnett’s (Multiple Range) test was used to assess the significance of inter-group differences; identifying differences of <0.05 or <0.01 as appropriate.

In case of the SPSS PC+4.0 program package, the heterogeneity of variance between groups was checked by Bartlett's test. Where no significant heterogeneity was detected, a one-way analysis of variance (ANOVA) was carried out. If the obtained result was significant, then Duncan's Multiple Range test was used to assess the significance of inter-group differences.
Indices:
- Number of corpora lutea: mean ± S.D.
- Number of implantations: mean ± S.D.
- Number and percentage of live foetuses: mean ± S.D.
- Number and percentage of intrauterine mortality: mean ± S.D.
Classified according to time of death: preimplantation loss, postimplantation loss, early and late embryonic loss, as well as foetal death

Foetal Data:
- Sex distribution: %, group mean
- Foetal body weight : mean ± S.D.
- External abnormalities/litter: %, group mean
- Visceral abnormalities/litter: %, group mean
- Skeletal abnormalities/litter: %, group mean

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
A minor, but statistically significant (p<0.01) difference was noted in the body weight gain of the High dose group in GD 8-10 period when compared to the control, in the period GD 6-8 a similar low weight gain was observed, but was not statistically significant. From GD 10 the weight gain for this group was comparable with controls. This was a minor, transient effect with little or no consequences on the overall body weight values. It is not unusual to have such transient effects in the first few days of treatment, with full recovery after animals acclimatise to the test item. There was no effect on weight gain for the Mid or Low dose groups. The difference at the high dose was so small and transient, with no consequences on the overall body weight, and no statistical difference on the overall weight gain, that the small change is not considered as a clear adverse effect.

see Table 2 in the attachments
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Reduced daily food consumption was recorded for the High dose group at the beginning of the treatment (by 9% in the GD 6-8 period and by 13% in the GD 8-10 period. This transient difference was statistically significant at p<0.01) when compared to the control values (smaller changes without statistical significance were also recorded in the Mid dose group). The minor, transient difference at the high dose corresponds to the body weight gain data. The overall mean daily food consumption values in the treatment period or entire study did not differ more the 6% from the control values in any test item treated groups and there was no dose response in any case. Similar trends were seen in the total food consumption and food utilization parameters. Overall, no test item related effect on the food consumption was observed in the Mid or Low dose groups; a minor, transient reduced food intake in the High dose group was seen. The effect at the High dose was so small and transient with no significant on the overall study food intake that the difference is not considered to be adverse.
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:
no effects observed

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
see Table 4 in the attachments
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
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:
no effects observed

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: no adverse effects observed

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
see Table 5 in the attachments
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): see Table 5 in the attachments
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
see Table 5 in the attachments
Changes in sex ratio:
no effects observed
Description (incidence and severity):
see Table 5 in the attachments
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
see Table 5 in the attachments
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Description (incidence and severity):
see Table 6 in the attachments
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
see Table 10 in the attachments
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
see Table 8 in the attachments
Other effects:
no effects observed

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Applicant's summary and conclusion

Conclusions:
In this developmental toxicity study the test substance did not exhibit any adverse toxic effects to maternal animals and to foetuses up to and including the highest dose level of 1000 mg/kg/day.