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Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
250 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
GLP study conducted in accordance with OECD guideline therefore Klimisch 1 study.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

In the combined repeated dose toxicity study with the reproduction/developmental toxicity screening test in rats, the test substance Structural Analogue 04 (SA04) administered daily by oral gavage to Wistar rats did not result in test item related mortality or systemic adverse effects.

There were no statistically significant differences between the Control and test item-treated groups with regard to reproductive ability or in the mating or gestation indices, or effects considered adverse or toxicologically significant in correlation with the test substance administration. Test item administration was considered to have no impact on the duration of the mating period. Successful coitus (sperm positive vaginal smears and/or vaginal plugs) generally occurred within up to 6 days of pairing (cohabitation). No test item effect on the duration of pregnancy or abnormalities in the gestation outcome ascribed to the treatment were observed. No external abnormalities ascribed to treatment were detected at the clinical or external macroscopic examinations of the pups.

In this study, at the 1000 mg/kg bw/day dose level, the intrauterine mortality, post-natal mortality and subsequently the total mortality were higher than control. However, only the post-natal mortality attained statistical significance. These effects were considered to be possibly related to the test substance administration although a role of the individual variability could not be excluded.

At 1000 mg/kg bw/day, the mean number of viable pups was -26.1% lower than control on PND0 and -30.4% lower on PND4, attaining statistical significance when evaluated for the total number of pups. These results correlated with the higher post-natal and total mortality observed in the High dose dams.

There were no effects considered adverse on the offspring weight or weight gain at up to and including 250 mg/kg bw/day. At 1000 mg/kg bw/day, when evaluated per litter basis, there were no statistically significant differences to control, however, the mean litter weights were lower than control on both PND0 and PND4. When evaluated for all pups, the mean body weights were statistically significant, slightly lower up to -7% than control: on PND0, 6.39 g vs. 6.88 g, and on PND4, 10.36 g vs. 11.03 g. As the differences were minor and the dose response was not clear, a test item related adverse effect on the offspring development following administration of 1000 mg/kg bw/day to the parental generation in the conditions of this study could not be ascertained, although it could not be excluded.

Under the conditions of this study, based on histopathological changes in the kidney and stomach, the no observed adverse effect level (NOAEL) for the test substance for parental toxicity effects is considered to be 62.5 mg/kg bw/day. For reproductive toxicity, the no observed effect level (NOEL) for parental/adult effects is considered to be 1000 mg/kg bw/day for the males and 250 mg/kg bw/day for the females, due to the higher intrauterine, postnatal and total foetal mortality at 1000 mg/kg bw/day, correlated with a NOEL of 250 mg/kg bw/day for F1 offspring, based on a possible test substance-related slight effect on pups survival and birth weight at 1000 mg/kg bw/day.

In the combined repeated dose toxicity study with the reproduction/developmental toxicity screening test in rats, the test substance Structural Analogue 05 (SA05) administered daily by oral gavage to Wistar rats did not result in test item related mortality or systemic adverse effects.

SA05 administered to parental generation under the conditions of this study did not lead to any adverse effects considered related to treatment or toxicologically significant in the F1 generation at up to and including 250 mg/kg bw/day dose level. At 1000 mg/kg bw/day, the intrauterine mortality, post-natal mortality and subsequently the total mortality were statistically higher than control (28.06% vs. 9.73%, 8.12% vs. 0.61% and 32.7% vs. 10.3% in the High dose vs. the control groups), effects considered possibly related to SA05 administration at the High dose level.

There were no test substance-related adverse effects on the reproductive ability or in the mating or gestation indices. In all groups, the mating indices were 100%, the fertility indices, 92% due to 1/12 non-pregnant females in each group, and the gestation indices, 100%, values within the physiological ranges for Wistar rats. Successful coitus (sperm positive vaginal smears and/or vaginal plugs) generally occurred within up to 5 days of pairing (cohabitation). No test item effect on the duration of pregnancy or abnormalities in the gestation outcome ascribed to the treatment were observed. The mean duration of pregnancy was similar in the control and test item treated groups. There was no evidence of test substance-related macroscopic or microscopic findings in the High Dose animals in the reproductive organs. The number of corpora lutea and implantation sites were comparable in the treated groups up to and including 1000 mg/kg bw/day with the mean value recorded in the Control group.

For reproductive toxicity, the no observed effect level (NOEL) for parental/adult effects is considered to be 1000 mg/kg bw/day for the males and 250 mg/kg bw/day for the females, due to the higher intrauterine, postnatal and total foetal mortality at 1000 mg/kg bw/day, correlated with a NOEL of 250 mg/kg bw/day for F1 offspring, based on a possible test substance-related effect on pups survival at 1000 mg/kg bw/day.


Short description of key information:
A Reproduction/Developmental Toxicity Screening Test with structural analogues led to slightly statistically higher intrauterine mortality, post-natal mortality and subsequently total mortality at 1000 mg/kg bw/day.

Justification for selection of Effect on fertility via inhalation route:
Oral route is considered the most appropriate route of exposure.

Justification for selection of Effect on fertility via dermal route:
Oral route is considered the most appropriate route of exposure.

Effects on developmental toxicity

Description of key information
In Reproduction/Developmental Toxicity Screening Tests and a teratogenicity study with structural analogues, no structural or developmental adverse effects were observed in the surviving pups at up to and including 1000 mg/kg bw/day
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
14 September to 25 November 1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to EU test guidance in compliance with GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes
Limit test:
yes
Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG
- Age at study initiation: 65 to 75 days
- Weight at study initiation: 193.1 +/- 14.2
- Fasting period before study: NA
- Housing: single
- Diet: Altromin 1310 ad libitum
- Water: tap water ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 22°C
- Humidity (%): 48 to 61%
- Air changes (per hr): 16 to 20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 14. Sep To: 25. Nov 1993
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
- Frequency of preparation: daily
- Administration: within 3 hours after preparation

VEHICLE
- Concentration in vehicle: 200 mg/kg nominal
- Amount of vehicle (if gavage): 5 ml/kg
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
-
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1:1
- Length of cohabitation: over night (3:30 pm to 7:30 am next day)
- Verification of same strain and source of both sexes: yes - own breeding facility
- Proof of pregnancy: sperm in vaginal smear, referred to as day 1 of pregnancy
Duration of treatment / exposure:
7. - 16. day of pregnancy
Frequency of treatment:
daily
Duration of test:
cesarean section on Day 21 of pregnancy
No. of animals per sex per dose:
20 to 24 mated females
Control animals:
yes
Details on study design:
- Dose selection rationale: test item was tolerated in the acute and subacute studies without adverse effects
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE : Yes
- Time schedule for examinations: weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/100 g body weight/day: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: all organs examined macroscopically
uterus - live and dead fetuses, resorption sites, placentas
ovaries - corpora lutea

OTHER:
- diameter of conceptuses undergoing resorption
- placenta weights
- presence of iron in uterus walls with ammonium sulphide to detect invisible implantation sites
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Placenta 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
- Body weight: Yes: all per litter
- Crown-rump length: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No data
Statistics:
comparison to actual control group and historical controls
MANOVA: body weight development, fetal weight, placental weight
PURI&SEN rank order test: food intake
Mantel-Haenszel's chi-squared test: live fetuses, intrauterine fetal death, number of implants, number of corpora lutea
multivariate analysis of variance: litter means of fetal weights, crown-rump length, placental weights
Fisher test: autopsy findings, body cross-sections, skelettal examination

Dams which had no live fetuses were excluded from the calculation of mean values and statistical evaluation
Indices:
No data
Historical control data:
Yes
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
black discoloration of feces and blue discoloration of urine due to excretion of dye
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: other:
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
only minor anomalies or variation within the historical range of spontaneous findings were observed
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
No adverse effects were observed after daily administration of 1000 mg/kg/day in dams or their fetuses.
Maternal NOAEL: 1000 mg/kg/day
Fetal NOEL: 1000 mg/kg/day
Executive summary:

In this limit test, the test substance, dissolved in distilled water, was administered orally by stomach tube in a single daily dose of 1000 mg/kg body weight to a group of 20 pregnant female Wistar rats from the 7th - 16th day of pregnancy. A simultaneous control group of the same size received the vehicle without test compound. On the 21st day of pregnancy, the dams were killed and delivered by caesarean section. The foetuses delivered by caesarean section were then examined morphologically for developmental disorders.

The study showed that the repeated oral administration of the test substance at a dose level of 1000 mg/kg body weight in the sensitive phase of organogenesis for the conceptuses, did not lead to any impairment of the general physical condition of the dams or impaired intrauterine development of conceptuses.

The morphological examination of the foetuses with regard to stage of development, outwardly detectable anomalies as well as anomalies of the internal organs and the skeleton showed no indication of an embryotoxic or teratogenic effect of the compound. The findings observed are to be regarded as spontaneous in origin.

On the basis of the results of this limit test, the “no observed adverse effect level” in rats following oral administration lies at 1000 mg/kg body weight with regard to maternal and embryofoetal toxicity and teratogenicity.

No teratogenic effect was observed.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
GLP study conducted in accordance with OECD guideline therefore Klimisch 1 study.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

In this limit test, the test substance, dissolved in distilled water, was administered orally by stomach tube in a single daily dose of 1000 mg/kg body weight to a group of 20 pregnant female Wistar rats from the 7th - 16th day of pregnancy. A simultaneous control group of the same size received the vehicle without test compound. On the 21st day of pregnancy, the dams were killed and delivered by caesarean section. The foetuses delivered by caesarean section were then examined morphologically for developmental disorders.

The study showed that the repeated oral administration of the test substance at a dose level of 1000 mg/kg body weight in the sensitive phase of organogenesis for the conceptuses, did not lead to any impairment of the general physical condition of the dams or impaired intrauterine development of conceptuses.

The morphological examination of the foetuses with regard to stage of development, outwardly detectable anomalies as well as anomalies of the internal organs and the skeleton showed no indication of an embryotoxic or teratogenic effect of the compound. The findings observed are to be regarded as spontaneous in origin.

On the basis of the results of this limit test, the “no observed adverse effect level” in rats following oral administration lies at 1000 mg/kg body weight with regard to maternal and embryofoetal toxicity and teratogenicity.

No teratogenic effect was observed.


Justification for selection of Effect on developmental toxicity: via inhalation route:
Oral route is considered the most appropriate route of exposure.

Justification for selection of Effect on developmental toxicity: via dermal route:
Oral route is considered the most appropriate route of exposure.

Justification for classification or non-classification

The above studies have been ranked reliability 1 according to the Klimisch et al system. This ranking was deemed appropriate because the study was conducted to GLP and in compliance with agreed protocols. Sufficient dose ranges and numbers are detailed; hence it is appropriate for use based on reliability and animal welfare grounds.

The above results triggered no classification under the Dangerous Substance Directive (67/548/EEC) and the CLP Regulation (EC No 1272/2008). No classification for reproductive toxicity is therefore required.

Additional information