Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

According to REACH Regulation, Annex IX, 8.7, column 1, an Extended One-Generation Reproductive Toxicity Study shall be proposed, if the available repeated dose toxicity studies indicate adverse effects on reproductive organs or tissues or reveal other concerns in relation with reproductive toxicity. As no adverse effects on reproductive organs or tissues were detected in the available studies (28-day and 90-day studies, OECD TG 414), no further testing is required.

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available

Effects on developmental toxicity

Description of key information

NOEL(development)=250 mg/kg bw/d, NOAEL(maternal toxicity)=100 mg/kg bw/d

(rat, oral: gavage; OECD Guideline 414, GLP; dose levels: 0, 25, 100 and 250 mg/kg bw/day)

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 May 2016 and 15 July 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted 22 January 2001
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Version / remarks:
August 1998
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Version / remarks:
30 May 2008
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
San Juan
Remarks:
Sprague-Dawley Crl:CD (SD) IGS BR
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Weight at study initiation: 197 to 276 g
- Fasting period before study: no
- Housing: individually in solid-floor polypropylene cages with stainless steel mesh lids furnished with softwood flakes
- Diet (e.g. ad libitum): pelleted diet (Rodent 2018C Teklad Global Certified Diet, Envigo RMS (UK) Limited, Oxon, UK), ad libitum
- Water (e.g. ad libitum): mains drinking water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 ºC
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 20 May 2016 (first day of treatment) to 08 June 2016 (final day of necropsy)
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The stability and homogeneity of the test item formulations were previously determined by Envigo Research Limited, Shardlow, UK Analytical Services (Envigo Study Number: CQ79LS). Results showed the formulations to be stable for at least twenty-one days. Formulations were prepared on three occasions and stored at approximately +4 °C in the dark and under nitrogen.

VEHICLE
- Concentration in vehicle: 4.17, 16.7, 58.3/41.67 (Dose Level/Concentration reduced from the 26 May 2016 (Days 7 to 9 of gestation))
- Amount of vehicle (if gavage): 6 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples were taken of test item formulations and were analyzed for concentration of BADGE-MXDA at Envigo Analytical Laboratory, Shardlow. The results indicate that the prepared formulations were within ± 2% of the nominal concentration.
Details on mating procedure:
- Impregnation procedure: purchased timed pregnant (Animals were delivered in two batches containing females prior to Day 3 of gestation.)
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
Day 3 to Day 19 of gestation
Frequency of treatment:
daily
Duration of test:
18 d (aminals were sacrificed on day 20 of gestation)
Dose / conc.:
25 mg/kg bw/day (actual dose received)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
350 mg/kg bw/day (actual dose received)
Remarks:
Dose Level reduced to 250 mg/kg bw/d from the 26 May 2016 (Days 7 to 9 of gestation)
No. of animals per sex per dose:
24
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were chosen in collaboration with the Study Monitor and were based on previous toxicity data (Envigo Research Limited Study Number DP46NV).
- Rationale for animal assignment (if not random): The animals were randomly allocated to treatment groups using a randomization procedure based on stratified body weight to ensure similarity between the treatment groups.
Maternal examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: all animals were examined for overt signs of toxicity, ill-health or behavioral changes once daily during the gestation period; during the dosing period, observations were recorded immediately before dosing, up to thirty minutes after dosing and one hour after dosing

BODY WEIGHT: Yes
- Time schedule for examinations: Day 3 (before the start of treatment) and on Days 4, 5, 8, 11, 14 and 17 of gestation. Body weights were also recorded for surviving animals at terminal kill (Day 20).

FOOD CONSUMPTION: Yes
Food consumption was recorded for each surviving individual animal at Day 3, 5, 8, 11, 14, 17 and 20 of gestation

WATER CONSUMPTION: Yes
- Time schedule for examinations: daily

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #20
- Organs examined: ovaries and uteri
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: placental weight
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: all per litter
Statistics:
The following parameters were analyzed statistically, where appropriate, using the test methods outlined below:
Female body weight change, food consumption and gravid uterus weight: Shapiro Wilk normality test and Bartlett’s test for homogeneity of variance and one way analysis of variance, followed by Dunnett’s multiple comparison test or, if unequal variances were observed, on alternative multiple comparison test.
All caesarean necropsy parameters and fetal parameters: Kruskal-Wallis non-parametric analysis of variance; and a subsequent pairwise analysis of control values against treated values using the Mann-Whitney ‘U’ test, where significance was seen.
Fetal evaluation parameters, including skeletal or visceral findings: Kruskal-Wallis non-parametric analysis of variance and Mann-Whitney ‘U’ test.
Historical control data:
attached as .pdf file
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Noisy respiration and increased salivation were evident in the majority of females at the high dose level throughout the treatment period. In addition, decreased respiratory rate, labored respiration and hunched posture was evident in one surviving female on Day 8, decreased respiratory rate and labored/gasping respiration was evident in one surviving female on Day 10 and diarrhoea was evident in one surviving female on Day 17. One female also had generalized fur loss on days 19 and 20.
All of the females that were sacrificed in extremis showed increased salivation and respiratory pattern changes (noisy respiration, gasping/labored respiration and/or decreased respiratory rate). In addition, the female that was sacrificed in extremis on Day 5 had chromodacryorrhea, hunched posture, pilo-erection and lethargy, the female that was sacrificed in extremis on Day 7 had lethargy, hunched posture, pilo-erection and a distended abdomen, one of the females that was sacrificed in extremis on Day 14 had hunched posture and the other female that was sacrificed in extremis on Day 14 had hunched posture, a distended abdomen and was dehydrated. The female that was sacrificed in extremis on Day 16 also had lethargy and a mass under the left forelimb.
Instances of noisy respiration were evident in seven females treated with 100 mg/kg bw/day on Days 7, 12, 14, 15 or 17 and one female treated with 100 mg/kg bw/day had increased salivation on Day 12 only.
No such effects were detected in females treated with 25 mg/kg bw/day.
One control female had diarrhoea on Day 16 only.
Mortality:
mortality observed, treatment-related
Description (incidence):
Two females from the high dose group were sacrificed in extremis on Days 5 and Day 7 of gestation due to excessive body weight losses and adverse clinical signs. Following the reduction of the high dose level to 250 mg/kg bw/day a further three females were sacrificed in extremis on Days 14 and 16 and two females were found dead on Day 18 of gestation.
There were no further unscheduled deaths.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Females from the high dose group showed a reduction in body weight gain between Days 3 and 5 of gestation, with the majority of females showing actual body weight losses during this period. Two of these females were subsequently terminated on Days 5 and 7 due to excessive body weight loss. Following the reduction of the high dose level, improvement in body weight gain was evident in the surviving females however further body weight losses were evident in two females that were prematurely terminated and in the two females which were found dead. Body weight gain in the surviving high dose females was comparable to controls between Days 11 and 20. Cumulative body weight gain was statistically significantly reduced (p<0.05-0.01) in surviving females throughout the treatment period and body weight gain when adjusted for gravid uterus weight was also statistically significantly reduced (p<0.01) when compared to controls.
An initial reduction in body weight gain was evident in females treated with 100 mg/kg bw/day between Days 3 and 4 of gestation, with actual body weight losses being evident in some females. As a consequence of the initial reduction in body weight gain a slight reduction in cumulative body weight gain (achieving statistical significance between Days 3 and 14 only; p<0.05) was evident in these females and body weight gain when adjusted for gravid uterus weight was also slightly lower than controls. However statistical significance was not achieved for body weight gains or for body weight gain when adjusted for gravid uterus weight and body weight gains from Day 4 onwards were comparable to controls.
Body weight gain during gestation, including after adjustment for the contribution of the gravid uterus, was considered to be unaffected by treatment at 25 mg/kg bw/day.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Females from the high dose group showed a statistically significant reduction (p<0.01-0.001) in food consumption between Days 3 and 11. Food consumption between Days 11 and 20 was comparable to controls. Females treated with 100 mg/kg bw/day also showed a statistically significant reduction (p<0.01) in food consumption between Days 3 and 5. Recovery in these females was evident thereafter.
No differences as compared to the control group were detected on food consumption in females treated with 25 mg/kg bw/day.
Females treated with 100 mg/kg bw/day showed a statistically significant reduction (p<0.05) in food consumption between Days 11 and 14 however in the absence of a similar effect in the high dose females during this period the intergroup difference was most likely to represent biological variation.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Daily visual inspection of water bottles did not reveal any overt intergroup differences.
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:
effects observed, treatment-related
Description (incidence and severity):
All of the females from the high dose group that were sacrificed in extremis and one of the females that was found dead had gaseous distension in the stomach and gastro-intestinal tract. The female that was sacrificed in extremis on Day 7 also had a distended abdomen and the female that was sacrificed in extremis on Day 16 had a mass under the left forelimb that was filled with a pale brown viscous liquid. The remaining female that was found dead had a fluid filled thoracic cavity.
No macroscopic abnormalities were detected in the surviving females of the high dose group or in females treated with 100 or 25 mg/kg bw/day.
Neuropathological findings:
not examined
Histopathological findings: neoplastic:
not examined
Number of abortions:
no effects observed
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:
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): not examined
Changes in number of pregnant:
no effects observed
Details on maternal toxic effects:
There was no obvious effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), or pre- or post-implantation losses at 25, 100 or 350/250 mg/kg bw/day.
Intergroup differences for mean placental weights did not indicate any obvious effects of maternal treatment at 25, 100 or 350/250 mg/kg bw/day.
Statistical analysis of the data did not reveal any significant intergroup differences.
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
A statistically significant reduction (p<0.05-0.01) in the number of fetuses/litters showing incomplete ossification of the parietal was evident in all treated groups. Group mean values were within historical control ranges and the observation of one variant at a lower incidence compared with controls is not significant when evaluated in isolation. In the absence of any particular pattern of abnormal skeletal development of skeletal structures affecting treated fetuses, the observation of one affected skeletal structure can be considered unlikely to represent true developmental abnormality. This takes account of this finding being seen regularly on this study type amongst control group fetuses.
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
There was no obvious effect of maternal treatment on litter data as assessed by live litter size and sex ratio at 25, 100 or 350/250 mg/kg bw/day.
Intergroup differences for mean fetal and litter weights did not indicate any obvious effects of maternal treatment at 25, 100 or 350/250 mg/kg bw/day.
Neither the type, incidence nor the distribution of findings observed during external examination of the fetuses at necropsy on gestation Day 20 and subsequent detailed visceral and skeletal examination indicated any adverse effect of maternal exposure on fetal development.
A statistically significant reduction (p<0.05-0.01) in the number of fetuses/litters showing incomplete ossification of the parietal was evident in all treated groups. Group mean values were within historical control ranges and the observation of one variant at a lower incidence compared with controls is not significant when evaluated in isolation. In the absence of any particular pattern of abnormal skeletal development of skeletal structures affecting treated fetuses, the observation of one affected skeletal structure can be considered unlikely to represent true developmental abnormality. This takes account of this finding being seen regularly on this study type amongst control group fetuses.
Statistical analysis of the remaining data did not reveal any significant intergroup differences

Dose descriptor:
NOEL
Effect level:
250 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no effects observed
Abnormalities:
no effects observed
Developmental effects observed:
no

results tables are attached as .pdf file below

Conclusions:
The oral (gavage) administration of BADGE-MXDA to pregnant rats during gestation at dose levels of 25, 100 and 350 mg/kg bw/day (reduced to 250 mg/kg bw/day from Days 7 to 9 of gestation), resulted in treatment related effects at 350/250 mg/kg bw/day. The reduction in body weight gain and reduction in food consumption together with adverse clinical signs detected when the females were initially dosed at 350 mg/kg bw/day, resulted in the early sacrifice of two females (Day 5 and Day 7 of gestation). Following the reduction of the high dose level to 250 mg/kg bw/day, improvement was evident in some of the remaining females however a further three females were sacrificed early on Days 14 and 16 of gestation and two females were found dead on Day 18 of gestation. Although an initial reduction in body weight gain was noted for the 100 mg/kg bw/day dose group, full recovery was evident thereafter and only minimal clinical observations were evident. Consequently, 100 mg/kg bw/day was considered to represent the No Observed Adverse Effect Level (NOAEL) for the pregnant female.
No treatment-related changes were detected in the offspring parameters measured or on embryofetal development. The ‘No Observed Effect Level’ (NOEL) for developmental toxicity was therefore considered to be 250 mg/kg bw/day.
Executive summary:

The study was performed to investigate the effects of BADGE-MXDA on embryonic and fetal development following repeated administration by gavage to the pregnant female during gestation including the period of organogenesis. The study was designed to comply with OECD Guideline 414 (adopted 22 January 2001).

The test item was administered by gavage to three groups each of twenty-four time-mated Sprague-Dawley Crl:CD®(SD) IGS BR strain rats, between Days 3 and 19 of gestation inclusive at dose levels of 25, 100, and 350 mg/kg bw/day. Following the early termination of two females treated with 350 mg/kg bw/day and adverse clinical signs in the remaining high dose females, the high dose level was reduced to 250 mg/kg bw/day from the 26 May 2016 (Days 7 to 9 of gestation). A further group of twenty-four time-mated females was exposed to the vehicle only (Propylene Glycol) to serve as a control.

Clinical signs, body weight change, food and water consumptions were monitored during the study. 

All surviving females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weights, fetal weights, sex and external and internal macroscopic appearance were recorded. Half of each litter were examined for detailed skeletal development and the remaining half were subjected to detailed visceral examination.

…….

Mortality

Two females from the high dose group were sacrificedin extremison Days 5 and Day 7 of gestation due to excessive body weight losses and adverse clinical signs. Following the reduction of the high dose level to 250 mg/kg bw/day a further three females were sacrificed in extremis on Days 14 and 16 and two females were found dead on Day 18 of gestation. There were no further unscheduled deaths.

Clinical Observations

Noisy respiration and increased salivation were evident in the majority of females at the high dose level throughout the treatment period. Isolated incidences of decreased respiratory rate, gasping/labored respiration, hunched posture, diarrhoea or generalized fur loss were also evident in four surviving females from this dose group. In addition to increased salivation and respiratory pattern changes the females that were sacrificedin extremisalso showed incidences of chromodacryorrhea, hunched posture, dehydration, pilo-erection, lethargy, distended abdomen and/or a mass under the left forelimb. 

Instances of noisy respiration were also evident in females treated with 100 mg/kg bw/day and one female treated with 100 mg/kg bw/day had increased salivation on Day 12 only.

No such effects were detected in females treated with 25 mg/kg bw/day.

Body Weight

Females from the high dose group showed a reduction in body weight gain between Days 3 and 5 of gestation, with actual body weight losses being evident in some females. Following the reduction of the high dose level, improvement in body weight gain was evident in the surviving females however further body weight losses were evident in the females that were prematurely terminated on Days 14 and 16 or which were found dead on Day 18. Body weight gain in the surviving high dose females was comparable to controls between Days 11 and 20. Cumulative body weight gain was reduced in these females throughout the treatment period and body weight gain when adjusted for gravid uterus weight was also reduced when compared to controls.

A reduction in body weight gain was evident in females treated with 100 mg/kg bw/day between Days 3 and 4 of gestation, with actual body weight losses being evident in some females. Recovery was however evident thereafter. No such effects were evident in females treated with 25 mg/kg bw/day.

Food Consumption

Females from the high dose group showed a reduction in food consumption between Days 3 and 11. Food consumption between Days 11 and 20 was comparable to controls. Females treated with 100 mg/kg bw/day also showed a reduction in food consumption between Days 3 and 5. Recovery was evident thereafter. No differences as compared to the control group were detected on food consumption in females treated with 25 mg/kg bw/day.

Water Consumption

No differences as compared to the control group were detected on water consumption.


 

Post Mortem Studies

All of the females from the high dose group that were sacrificedin extremisand one of the females that was found dead had gaseous distension in the stomach and gastro-intestinal tract. The female that was sacrificedin extremison Day 7 also had a distended abdomen and the female that was sacrificedin extremison Day 16 had a mass under the left forelimb that was filled with a pale brown viscous liquid. The remaining female that was found dead had a fluid filled thoracic cavity. No treatment related macroscopic abnormalities were detected in the surviving females of the high dose group or in females treated with 100 or 25 mg/kg bw/day.

Litter Data and Litter Placental and Fetal Weights

The number of implantations, subsequent embryofetal survival, live litter size and sex ratio on Day 20 of gestation were considered to be unaffected by maternal treatment at 25, 100 or 350/250 mg/kg bw/day. Mean fetal, placental and litter weights were also considered to have been unaffected by maternal treatment at 25, 100 or 350/250 mg/kg bw/day.

Fetal Examination

External examination of fetuses on Day 20 of gestation did not indicate any obvious effect of maternal treatment on fetal development at 25, 100 or 350/250 mg/kg bw/day. Findings at detailed skeletal and visceral examinations of fetuses on Day 20 of gestation did not indicate any obvious effect of maternal treatment on fetal development at 25, 100 or 350/250 mg/kg bw/day.

Conclusion

The oral (gavage) administration of BADGE-MXDA to pregnant rats during gestation at dose levels of 25, 100 and 350 mg/kg bw/day (reduced to 250 mg/kg bw/day from Days 7 to 9 of gestation), resulted in treatment related effects at 350/250 mg/kg bw/day. The reduction in body weight gain and reduction in food consumption together with adverse clinical signs detected when the females were initially dosed at 350 mg/kg bw/day, resulted in the early sacrifice of two females (Day 5 and Day 7 of gestation). Following the reduction of the high dose level to 250 mg/kg bw/day, improvement was evident in some of the remaining females however a further three females were sacrificed early on Days 14 and 16 of gestation and two females were found dead on Day 18 of gestation. Although an initial reduction in body weight gain was noted for the 100 mg/kg bw/day dose group, full recovery was evident thereafter and only minimal clinical observations were evident. Consequently, 100 mg/kg bw/day was considered to represent the No Observed Adverse Effect Level (NOAEL) for the pregnant female.

No treatment-related changes were detected in the offspring parameters measured or on embryofetal development. The ‘No Observed Effect Level’ (NOEL) for developmental toxicity was therefore considered to be 250 mg/kg bw/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
250 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
guideline study, GLP, reliable without restriction
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

A study was performed to investigate the effects of BADGE-MXDA on embryonic and fetal development following repeated administration by gavage to the pregnant female during gestation including the period of organogenesis. The study was designed to comply with OECD Guideline 414 (adopted 22 January 2001).

The test item was administered by gavage to three groups each of twenty-four time-mated Sprague-Dawley Crl:CD®(SD) IGS BR strain rats, between Days 3 and 19 of gestation inclusive at dose levels of 25, 100, and 350 mg/kg bw/day. Following the early termination of two females treated with 350 mg/kg bw/day and adverse clinical signs in the remaining high dose females, the high dose level was reduced to 250 mg/kg bw/day from the 26 May 2016 (Days 7 to 9 of gestation). A further group of twenty-four time-mated females was exposed to the vehicle only (Propylene Glycol) to serve as a control.

Clinical signs, body weight change, food and water consumptions were monitored during the study. 

All surviving females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weights, fetal weights, sex and external and internal macroscopic appearance were recorded. Half of each litter were examined for detailed skeletal development and the remaining half were subjected to detailed visceral examination.

…….

Mortality

Two females from the high dose group were sacrificedin extremison Days 5 and Day 7 of gestation due to excessive body weight losses and adverse clinical signs. Following the reduction of the high dose level to 250 mg/kg bw/day a further three females were sacrificedin extremison Days 14 and 16 and two females were found dead on Day 18 of gestation. There were no further unscheduled deaths.

Clinical Observations

Noisy respiration and increased salivation were evident in the majority of females at the high dose level throughout the treatment period. Isolated incidences of decreased respiratory rate, gasping/labored respiration, hunched posture, diarrhoea or generalized fur loss were also evident in four surviving females from this dose group. In addition to increased salivation and respiratory pattern changes the females that were sacrificedin extremisalso showed incidences of chromodacryorrhea, hunched posture, dehydration, pilo-erection, lethargy, distended abdomen and/or a mass under the left forelimb. 

Instances of noisy respiration were also evident in females treated with 100 mg/kg bw/day and one female treated with 100 mg/kg bw/day had increased salivation on Day 12 only.

No such effects were detected in females treated with 25 mg/kg bw/day.

Body Weight

Females from the high dose group showed a reduction in body weight gain between Days 3 and 5 of gestation, with actual body weight losses being evident in some females. Following the reduction of the high dose level, improvement in body weight gain was evident in the surviving females however further body weight losses were evident in the females that were prematurely terminated on Days 14 and 16 or which were found dead on Day 18. Body weight gain in the surviving high dose females was comparable to controls between Days 11 and 20. Cumulative body weight gain was reduced in these females throughout the treatment period and body weight gain when adjusted for gravid uterus weight was also reduced when compared to controls.

A reduction in body weight gain was evident in females treated with 100 mg/kg bw/day between Days 3 and 4 of gestation, with actual body weight losses being evident in some females. Recovery was however evident thereafter. No such effects were evident in females treated with 25 mg/kg bw/day.

Food Consumption

Females from the high dose group showed a reduction in food consumption between Days 3 and 11. Food consumption between Days 11 and 20 was comparable to controls. Females treated with 100 mg/kg bw/day also showed a reduction in food consumption between Days 3 and 5. Recovery was evident thereafter. No differences as compared to the control group were detected on food consumption in females treated with 25 mg/kg bw/day.

Water Consumption

No differences as compared to the control group were detected on water consumption.


 

Post Mortem Studies

All of the females from the high dose group that were sacrificedin extremisand one of the females that was found dead had gaseous distension in the stomach and gastro-intestinal tract. The female that was sacrificedin extremison Day 7 also had a distended abdomen and the female that was sacrificedin extremison Day 16 had a mass under the left forelimb that was filled with a pale brown viscous liquid. The remaining female that was found dead had a fluid filled thoracic cavity. No treatment related macroscopic abnormalities were detected in the surviving females of the high dose group or in females treated with 100 or 25 mg/kg bw/day.

Litter Data and Litter Placental and Fetal Weights

The number of implantations, subsequent embryofetal survival, live litter size and sex ratio on Day 20 of gestation were considered to be unaffected by maternal treatment at 25, 100 or 350/250 mg/kg bw/day. Mean fetal, placental and litter weights were also considered to have been unaffected by maternal treatment at 25, 100 or 350/250 mg/kg bw/day.

Fetal Examination

External examination of fetuses on Day 20 of gestation did not indicate any obvious effect of maternal treatment on fetal development at 25, 100 or 350/250 mg/kg bw/day. Findings at detailed skeletal and visceral examinations of fetuses on Day 20 of gestation did not indicate any obvious effect of maternal treatment on fetal development at 25, 100 or 350/250 mg/kg bw/day.

Conclusion

The oral (gavage) administration of BADGE-MXDA to pregnant rats during gestation at dose levels of 25, 100 and 350 mg/kg bw/day (reduced to 250 mg/kg bw/day from Days 7 to 9 of gestation), resulted in treatment related effects at 350/250 mg/kg bw/day. The reduction in body weight gain and reduction in food consumption together with adverse clinical signs detected when the females were initially dosed at 350 mg/kg bw/day, resulted in the early sacrifice of two females (Day 5 and Day 7 of gestation). Following the reduction of the high dose level to 250 mg/kg bw/day, improvement was evident in some of the remaining females however a further three females were sacrificed early on Days 14 and 16 of gestation and two females were found dead on Day 18 of gestation. Although an initial reduction in body weight gain was noted for the 100 mg/kg bw/day dose group, full recovery was evident thereafter and only minimal clinical observations were evident. Consequently, 100 mg/kg bw/day was considered to represent the No Observed Adverse Effect Level (NOAEL) for the pregnant female.

No treatment-related changes were detected in the offspring parameters measured or on embryofetal development. The ‘No Observed Effect Level’ (NOEL) for developmental toxicity was therefore considered to be 250 mg/kg bw/day.

Justification for classification or non-classification

Based on reliable, relevant and adequate data the substance does not need to be classified for toxicity to reproduction, developmental toxicity and teratogenicity according to the criteria given in regulation (EC) 1272/2008. No treatment-related changes were detected in the offspring parameters measured or on embryofetal development. Therefore labelling is not necessary.