Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
January-February 2009
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: (as the data is used in a read-across approach, a maximal reliability score of 2 was attributed).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes (incl. certificate)
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: LAB 3822,
Supplier: ROQUETTE FRERES,
Batch Number: FAL 07/25,
Galenic form: Slightly yellowish liquid,
Molecular weight (base form): about 426 g/mol,
Expiry date (corresponding to shelf life on the certificate of analysis): March 2009 extended to March 2010, according to the last analysis,
Intended used: Ind. Chemical.

Storage conditions: Immediately upon receipt, the test item was registered, then stored at ambient temperature.
Handling instructions for LAB 3822: General safety procedures as appropriate for handling of chemicals of unknown hazard potential were applied.
Incompatibility: No suspected incompatibilities of the test item with any material likely to come in contact with it during the course of the study were specified by the Sponsor.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
Identification: Animals were identified individually by an ear clip.
Number: 116 mated females, in order to obtain at least 80 pregnant females, in the main study and 12 pregnant females in the satellite groups.
Age: 10 to 11 weeks at the beginning of the treatment period.
Weight: Between 214.2 g and 296.3 g on the day of randomisation with a maximum range of 82.1 g.
Spare animals: Spare animals were kept under the same environmental conditions and may replace any animal in case of unforeseen events. At the end of the study period, all spare animals remaining unassigned were released from the study.
Acclimatisation: Animals arrived at CERB on Day 1 of pregnancy (D1pc). Animals were supplied in 8 batches for logistical reason.
At least five days in the laboratory animal house where the experiment took place. Only animals without any visible sign of illness were used for the study.
Housing: Daily observations were undertaken at the time of delivery of the animals and during the period of acclimatisation. Animals were housed individually in cages of standard dimensions (375-215-149 mm) with sawdust bedding (or equivalent).
The animals were placed in an air-conditioned (19-23°C) animal house kept at relative humidity between 45% and 65% (except during the cleaning slot) in which non-recycled filtered air was changed approximately 10 times per hour. The artificial day/night cycle involved 12 hours light and 12 hours
darkness with light on at 7.30 a.m..
Feeding: RM3 (E)-SQC SDS/DIETEX feed (quality controlled/radiation sterilised) was available ad libitum except during the fasting experimental period.The certificate of analysis concerning this feed product is included in Appendix C, page 183. The criteria for acceptable levels of contaminants in the
feed supplied were within the limits of the analytical specifications established by the diet manufacturer.
Drinking water: Drinking water was available ad libitum in polycarbonate bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to the LAEASE Région Sud Est, 5, avenue Achille Maureau, B.P. 95, 84703 Sorgues Cedex, France, for analysis. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.

Administration / exposure

Route of administration:
oral: unspecified
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
The breeding establishment was responsible for mating. Females were mated at the beginning of the morning. They were inspected for the presence of a vaginal plug. The day on which a female was found to have a vaginal plug was considered to be Day 0 of pregnancy (D0).
Duration of treatment / exposure:
LAB 3822 or sterile water was administered once a day at each chosen dose level, by the oral route from D6 to D19 of pregnancy (i.e., from implantation to one day before termination).
Frequency of treatment:
daily
Duration of test:
From January 19 to February 12 2009.
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0.575 g/kg
Basis:

Remarks:
Doses / Concentrations:
1.15 g/kg
Basis:

Remarks:
Doses / Concentrations:
2.3 g/kg
Basis:

No. of animals per sex per dose:
The study involved 4 groups of 24 (group 2) and 25 (groups 1, 3, 4) SPF Sprague-Dawley rats (mated females) and 4 satellite groups (each of 4 mated females).
Control animals:
yes
Details on study design:
Proposed doses selected in agreement with the Sponsor are based on previous studies. The highest dose should reveal signs of toxicity and the lowest dose should represent a no-observed-adverse effect level (NOAEL). Proposed method of administration was selected since it is the intended mode of administration in Human.

Dose adjustment
Water content: about 0.1% (according to the certificat of analysis).
Doses of test item were expressed in g/kg of DEI (Di-ester-isosorbide) and were adjusted on the basis of the most recent body weight. A correction factor of 1.15 was used (1 g of LAB 3822 contains 87 % of DEI).

Examinations

Maternal examinations:
Mortality/morbidity was recorded twice a day.
General observations were performed before the first dosing (i.e., on D5) and once a day up to the end of the study at 60 min post dose (± 30 min). Animals were weighed on D1, D5 (day of randomisation), from D6 to D19 and on D20 (day of necropsy not fasted and not exsanguinated).
Food and water consumption were measured daily from D6 to D19.
Blood samples for drug analysis were taken from satellite animals. Four females were sampled per time point and per group on D6 and on D19 (before dosing and then at 2 and 4 hours post-dosing).
Females surviving at the end of the study were submitted to gross necropsy.
Ovaries and uterine content:
Uterus and ovaries from all females were observed macroscopically: the uterus was weighed before extraction of foetuses and fixed.
At the time of the caesarean, corpora lutea on ovaries, number and uterine location of implantation sites, resorptions (early and late), and foetuses (live and dead) were noted.
Fetal examinations:
Foetuses were examined macroscopically and the following were noted:
• Number of live or dead foetuses,
• Individual foetal body weight (live and dead),
• Caudo-cranial measurement of live and dead foetuses,
• Gross evaluation and weight of placenta of all foetuses,
• external morphological examination of these foetuses,

After euthanasia, half of the total number of foetuses from each litter were fixed in aqueous Bouin0s fluid for visceral examination and the other half were placed in 95% alcohol for skeletal examination.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
No mortality attributed to the treatment with the vehicle or with the test item was seen whatever the
dose.
No clinical signs was seen in animals dosed with sterile water or LAB 3822.
There was no difference in body weight gain between the control group and LAB 3822 treated groups.
Food and water consumption were slightly decreased in animals dosed with LAB3822 whatever the dose.
Macroscopic examinations:
No abnormality attributed to sterile water or to LAB 3822 at the doses of 0.5, 1 and 2 g of DEI/kg was noticed in any organs examined at necropsy.
Uterus weight:
No statistically significant change in uterus weight was seen in females dosed with LAB 3822 at any doses when compared to the animals dosed with the vehicle.
Pre-implantation loss, number of live foetuses, post-implantation loss, foetal incidence:
No significant difference was seen between females dosed with LAB 3822 at the doses of 0.5, 1 and 2 g of DEI/kg and females dosed with the vehicle.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
> 500 - < 2 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Necropsy of foetuses:
No foetus was found dead at the time of caesarean of females dosed with sterile water or with LAB 3822 whatever the dose.
Except for haemorrhagic points observed in all dose-groups, no other external examination was observed.
No difference between foetal parameters was noted whatever the group.
Foetal examination (visceral and skeletal):
Examination of Bouin-fixed foetuses following freehand serial sectioning did not reveal any changes that were considered to be related to maternal treatment with LAB3822 at dose levels up to 2.0 g DEI/kg body weight/day.
Examination of alizarin stained foetal skeletal preparations indicated a marginal reduction in ossification parameters at 2.0 g DEI kg body weight/day which was a consequence of a slightly higher incidence of foetuses of low body weight(<3.0g) at this dose level. With this exception, there were no skeletal changes that were considered to be related to maternal treatment with LAB 3822.

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
LAB 3822 administered at the doses of 0.5, 1 and 2 g of DEI/kg, except a marginal reduction in ossification parameters at 2.0 g of DEI/kg, which was a consequence of a slightly higher incidence of foetuses of low body weight less than 3g, did not cause any effects on the embryo-foetal
development in the rat.