Dimethyl sebacate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

There is a GLP combined repeated dose toxicity stuidy in rats performed according to OECD 422. No effects were obseved on fertility or reproductive organs at the top dose-levels of 1000 mg/kg/day.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 january -13 May 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animal information
Species, strain, supplier Rat: HsdHan™:WIST from Harlan, Bicester, UK.
Specification At start of dosing: 10 to 12 weeks old.
Justification A readily available rodent species acceptable to the regulatory authorities and recommended for reproduction studies because of its reproductive characteristics.

Environment
Housing : Cages conforming to the 'Code of practice for the housing and care of animals used in scientific procedures' (Home Office, London, 1989).
Housing density: Groups of up to four (pre-pairing and post-pairing), one female with one male (pairing) or individually (mated females).

Relevant animals were housed individually for approximately 24 hours# prior to FOB assessment.
# During the mating period, any male or female that did not show evidence of mating on the day prior to their FOB assessment, were continued to be paired for mating. On the day of testing, the animal was removed from the mating cage and housed singly for approximately 1 hour before testing. After testing, animals were placed back into mating cages as required.
Rooms exclusive to study Yes
Target temperature range 20 to 24°C.
Target humidity range 45 to 65%.
Air changes 15 -20 air changes/hour.
Photo-period 12 hours nominal.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

Administration volume: 5 mL/kg. Individual dose volumes will be based on individual body weight.

Details on mating procedure:

One male was paired with one female of the same treatment group. Mating was confirmed by the presence of sperm in a vaginal smear or a retained vaginal plug. The day of confirmation of mating was taken as Day 0 of gestation.
Pairing took place after two weeks of treatment.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Formulations prepared at 10 and 200 mg/mL were analysed to determine homogeneity and stability. The formulations were to be considered be homogeneous if the coefficient of variation (CV) of the results was £ 6.0% and the homogeneity results were within ± 10% of the mean. The formulations were to be considered stable if the mean of the results at each time point were within ±10% of the mean at 0 hour. Stability of the formulations at the concentration of 10 mg/mL and 200 mg/mL for up to 10 days at room temperature (15 ºC to 25 ºC) was demonstrated. The results of the 10 mg/mL suspension formulated for the stability trial were low, but the mean of the results at each time point were within ±10% of the mean at 0 hour over the 10 day period evaluatedwith a CV of 0.33 to 0.72% (at 10 mg/mL).
Formulations prepared for use in Weeks 1, 3 and 6 of the study were analysed to determine homogeneity and achieved concentration. The formulations were to be considered be homogeneous if the coefficient of variation (CV) of the results was £ 6.0% and the homogeneity results were within ± 10% of the mean. The target range for the preparation of the formulations was 90 to 110% of nominal. Results were within the range 97 to 106%, with a CV of 0.30 to 1.43%, and therefore acceptable for dosing. Test article was not detected in the Group 1 control samples.

Duration of treatment / exposure:

The test article was administered orally by gavage.
Males were dosed once daily for two weeks prior to pairing, during the pairing period and a further two weeks before necropsy. The males were treated for at least six weeks.
Females were dosed once daily for two weeks prior to pairing, during the pairing period and until Day 4 post-partum inclusive. The females were allowed to litter and rear their offspring to Day 4 post-partum. Dosing was deferred or omitted if the animal was in or near parturition.

Frequency of treatment:

The dosage were admniistered daily.

Remarks:

Doses / Concentrations:
100, 300, 100 mg/kg/day
Basis:

No. of animals per sex per dose:

10 animals/sex/group

Control animals:

yes, concurrent vehicle

Details on study design:

Daily oral (gavage) administration of Dimethyl Sebacate at 100, 300 and 1000 mg/kg/day in a 14 Day oral (gavage) range finding study (Covance Study Number 8273520) was generally well tolerated, with no unscheduled deaths or remarkable clinical signs. There were no dose limiting post dosing observations. Mouth rubbing was noted immediately after dosing, throughout the majority of the dosing period in all male animals (including controls); this lasted for up to 2 hours post dose on one occasion at 1000 mg/kg/day. Mouth rubbing was also seen immediately after dosing in the majority of females (including controls), but on fewer occasions than males. Salivation and paddling was also observed post dosing in all groups, for up to 2 hours in males given Dimethyl Sebacate. There were no adverse effects of dosing on body weight, body weight gains or food consumption in males or females. At terminal necropsy, there were no adverse macroscopic findings or effects on organ weights. Therefore, based on these findings, the same dose levels were considered suitable for the Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test.

Parental animals: Observations and examinations:

Each animal was given a detailed physical examination at weekly intervals. An individual record was maintained of the clinical condition of each animal on days of body weight recording. Additional observations were recorded as deemed necessary.
Male animals were weighed on Day -7, then at weekly intervals and the day of (prior to) necropsy.

Body weight :
Females were weighed on Day 7, weekly prior to pairing and until confirmation of mating, Days 0, 7, 14 and 20 of gestation, Days 1 and 4 post partum and on the day of necropsy.

Food consumption :
Food consumption was recorded over the same intervals as the body weights for all males prior to and post pairing and females prior to pairing. In addition, food consumption was determined for females over Days 0 to 4, 4 to 7, 7 to 10, 10 to 14, 14 to 17 and 17 to 20 of gestation (reported over the same intervals as the body weights) and Days 1 to 4 post partum.



On Day -6, the male animals that were re-housed for FOBs were given food ad-lib in error. The food from the home cage should have been evenly distributed between each male. Therefore, the food consumption values for Week -1 showed that the animals appear to have eaten less.

Oestrous cyclicity (parental animals):

Estrous was examined during the pairing period until vaginal pug is observed.

Sperm parameters (parental animals):

no data

Litter observations:

Procedures for post-partum phase
All parental females were allowed to litter and rear offspring to Day 4 post partum. The day the pups were first observed was designated Day 0 post partum.
The date of parturition was recorded and the duration of gestation was calculated.
The following data were recorded for each litter:
¿ number of pups born (live and dead)
¿ daily live litter size and sex (reported on Days 1 and 4)
¿ daily clinical observations
¿ individual pup weights on Days 1 and 4 post-partum
In addition, daily records of mortality and changes in litter size were maintained. Where possible, pups found dead or moribund were given a gross necropsy

Postmortem examinations (parental animals):

All adult animals including decedents were subjected to necropsy.
The adult scheduled necropsies were performed after an overnight period without food. Where possible the scheduled necropsies were carried out in group order for all adult animals. Each adult was given isoflurane anaesthesia. Once a suitable deep plane of anaesthesia was established, the animal was exsanguinated by the severing of major blood vessels. A full macroscopic examination was performed under the general supervision of a pathologist and all lesions were recorded.
Pups were given an intraperitoneal injection of sodium pentobarbitone (overdose). Once a suitable deep plane of anaesthesia had been established, the animal was exsanguinated by the severing of major blood vessels.

The uterus of each littering female and any mated not pregnant female was immersed in a 10% ammonium sulphide solution to reveal any evidence of implantation.

The following tissues from each animal were preserved in 10% neutral buffered formalin unless otherwise indicated.

W = weighed; E = processed and examined microscopically

adrenals W E Peyer’s patch E
animal identification# pituitary E
aorta E popliteal lymph nodes E
bone marrow smear (femur) (1) (2) E prostate# W E
brain W E rectum E
caecum E sciatic nerves E
colon E seminal vesicles# W E
duodenum E spinal cord cervical E
femur with bone marrow and stifle joint E spinal cord lumbar E
gross lesions (variable)# E spinal cord thoracic E
heart W E spleen W E
ileum E sternum with bone marrow E
jejunum E stomach E
kidney W E testes and epididymides# (3) (4) W E
larynx E thymus W E
liver W E thyroids + parathyroids W E
lungs with mainstem bronchi and bronchioles E tongue E
mandibular lymph nodes W E trachea E
mesenteric lymph nodes E trachea bifurcation E
oesophagus E ureters E
ovaries# W E urinary bladder# E
oviducts E uterus including cervix# W E
pancreas E vagina# E
1 – see clinical pathology section (Section 3.7.3)
2 – tissue taken into bovine albumin; smear prepared; air dried, then fixed in methanol
3 - tissue taken into Bouin’s fixative and processed at least to block stage
4 – sections of testes were stained with Periodic Acid Schiff-Haematoxylin (PASH) for qualitative assessment of
spermatogenic stages
# - all adult animals
Bone designated for histopathological examination was decalcified using Kristenson’s fluid.

Adult animals were weighed before necropsy.
The following sexual organs of the five highest numbered main study animals/sex/group denoted by ‘W’ in the tissue list above were dissected free from fat and other contiguous tissue and weighed before fixation (the testes and epididymides were weighed for all animals). Left and right organs were weighed together. (seminal vesicles, testes, epididymides, ovaries, uterus)

Postmortem examinations (offspring):

none

Statistics:

Mating, fertility and fecundity indices were analysed using the Cochran-Armitage test for dose response and Fisher’s exact test for pairwise comparisons. The tests were interpreted with one-sided risk for decreased incidence with increasing dose. A significant trend (P<0.05) was only reported where none of the pairwise comparisons was significant.
The number of implantation sites, number of pups born, the percentage of male pups at Day 1, pup weights, post-implantation survival index and live birth index were analysed using non parametric methods. The non-parametric methods employed were the Kruskal-Wallis ANOVA, the Terpstra Jonckheere test for a dose related trend and the Wilcoxon rank sum test for pairwise comparisons. Where the Kruskal-Wallis ANOVA was not significant, the pairwise comparisons were not reported in order to protect the Type I error.
Some variables were not analysed due to all animals having the same value or there being too few distinct values for meaningful analysis.

Reproductive indices:

The following indices were calculated: mating index, female fecundity index, male fecundity index, male fertility index, post-implantantion survival index,

Offspring viability indices:

Gestation index, live birth index and viability index were calculated.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

not examined

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Mortality:
There were no deaths in males and females that could be attributed to the treatment with the test item. Unscheduled deaths during the study were limited to females killed due to their having total litter loss (see Litter Data).

Post-dosing observations:
For males, minor post-dosing observations such as mouth-rubbing, paddling and salivation were seen immediately post-dose or occasionally for longer from Day 9 or 12 of dosing throughout the dosing period on many occasions in the majority of animals receiving 1000 mg/kg/day, few animals receiving 300 mg/kg/day, rarely in animals receiving 100 mg/kg/day,. For females administered 1000 mg/kg/day, there was an isolated incident of mouth rubbing and salivation immediately post-dosing only on Day 12 and Day 13 of dosing in the pre-pairing period. During gestation for females that received 1000 mg/kg/day mouth rubbing was observed each day after dosing and was frequently associated with paddling and occasionally associated with salivation. Mouth rubbing and paddling were also seen for females that received 100 or 300 mg/kg/daybut the frequency and number of animals involved was lower.
These observations were considered to be indicative of taste aversion rather than systemic toxicity.

Body weight
The variations observed at the high dose-level were slight and not statistically significant. Therefore the slight decrease was not considered to be of biological significance.

Food consumption
There was no effect of Dimethyl Sebacate administration on mean food intake either before pairing in males and females or in females during gestation and lactation.



Mating data

Median pre-coital time, pregnancy rate, mating index, and male and female fertility and fecundity indexes were all unaffected by treatment.

Organ weights
Pathology Report
Organ weight and/or organ weight ratio changes, were attributed to normal biological variation and were considered not to be related to administration of Dimethyl Sebacate as they were either small in magnitude, not dose-dependent, inconsistent between the sexes, due to normal inter-animal variability and/or lacked a histopathological correlate.

Macroscopic Data
Pathology Report
There were no macroscopic findings suggestive of effects of Dimethyl Sebacate. Most tissues were macroscopically unremarkable and the findings seen were generally consistent with the usual pattern of findings in animals of this strain and age.

Microscopic Data
Pathology Report
There were no microscopic findings suggestive of effects related to administration of Dimethyl Sebacate. Most tissues were microscopically unremarkable and the findings seen were generally consistent with the usual pattern of findings in animals of this strain and age. Atrophy was recorded in the thymus of most female animals, this was considered to be related to the stress of pregnancy and/or birth, and not related to administration of Dimethyl Sebacate.
Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

no

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Litter data

After parturition, 2, 1, 1, and 2 females from Groups 1, 2, 3 and 4 respectively showed total litter loss. In 3 from 6 of the dams with total litter loss, at least one pup had the umbilical cord still attached after parturition; at least one litter mate was also still born which may indicate a more difficult parturition for these dams. The distribution of these losses is similar to that seen in previous littering studies conducted at these laboratories in the same strain of rat, and therefore was considered not to be an adverse effect of treatment. There was no effect of treatment on duration of gestation, number of implantation sites, number of pups born, sex ratio, the mean number of pups alive on Day 1 and Day 4 of age, post-implantation survival index, live birth index or viability index.

Mean pup weight : The mean pup weight was considered to be similar in all groups on Day 1 of lactation (Group 4 combined pup body weight was approximately 3 % lower than the control). On Day 4 mean pup weight was slghtly decreased at all dose-levels when compared to control, however as these valuiation were not dose-related, not statistiscally significant and within the range of laboratory historical control data, they were not considered to be treatment related.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

no

Reproductive effects observed:

no

Conclusions:

A combined repeated dose toxicity study with the reproduction dose toxicity study with the reproduction/developmental toxicity screening test in the rat was performed at dose-levels of 100, 300 and 1000 mg/kg/day. The changes observed were not considered of biological significance or related to the treatment. The NOAEL was 1000 mg/kg/day for general toxicity, fertility and pup development.

Executive summary:

The objective of the study (Rhodes, 2013) performed according to OECD 422 guideline was to provide preliminary information on the effects of the test article, Dimethyl Sebacate, following daily oral (gavage) administration to the rat and to provide a preliminary evaluation of reproduction/developmental toxicity.

Groups of rats of the HsdHan^TM:WIST strain were dosed for 2 weeks prior to pairing for males dosing continued during pairing until mating and then until the day before necropsy. For females dosing continued throughout pairing, gestation and until Day 4 of lactation at 100, 300 or 1000 mg/kg/day dose-levels.

Body weights, clinical signs, food consumption and functional observation battery testing were recorded during the treatment period. Blood samples were taken for Haematology and clinical chemistry and urine samples were collected and analysed.

Analysis trials performed on concentrations from 10 to 200 mg/mL showed that the formulated test material was stable and homogenous. Prepared dosing formulations were found to be in the target range for achieved concentration, and homogenous.

There were no treatment-related deaths, adverse clinical observations, or necropsy findings during the study. Minor post-dosing observations associated with taste-aversion were seen in many treated animals during the study

Group mean body weight gain was variable with high dose males gaining slightly less weight than controls over the course of the study but this was not dose-related and not statisically significant, therefore it was considered of no biological relevance. There was no clear effect of treatment on mean food intake either before pairing in males and females or in females during gestation.

Mating and fertility data showed no effect of treatment.

After parturition, 2, 1, 1, and 2 females from Groups 1, 2, 3 and 4 respectively showed total litter loss. The distribution of these losses is similar to that seen in previous littering studies conducted at these laboratories in the same strain of rat, and therefore was not considered to be an adverse effect of treatment. There was no effect of treatment on intrauterine data, or on group mean pup data, with the exception of Day 4 mean pup weights in both sexes which were slightly lower than controls but the changes were not statistically significant and within the range of historical control data, therefore it was not considered to be of toxicological relevance.
In conclusion, oral gavage administration of 1000 mg/kg/day Dimethyl Sebacate to male and female rats produced was characterised by slightly decreased body weight gain in males only.

The no-observed-adverse-effect-level (NOAEL) for pup developement was 1000 mg/kg/day.

Effect on fertility: 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:

The study is valid performed according to GLP and compliant with OECD 414.

Additional information

The objective of the study (Rhodes, 2013) performed according to OECD 422 guideline was to provide preliminary information on the effects of the test article, Dimethyl Sebacate, following daily oral (gavage) administration to the rat and to provide a preliminary evaluation of reproduction/developmental toxicity.

Groups of rats of the HsdHan^TM:WIST strain were dosed for 2 weeks prior to pairing for males dosing continued during pairing until mating and then until the day before necropsy. For females dosing continued throughout pairing, gestation and until Day 4 of lactation at 100, 300 or 1000 mg/kg/day dose-levels.

Body weights, clinical signs, food consumption and functional observation battery testing were recorded during the treatment period. Blood samples were taken for Haematology and clinical chemistry and urine samples were collected and analysed.

Analysis trials performed on concentrations from 10 to 200 mg/mL showed that the formulated test material was stable and homogenous. Prepared dosing formulations were found to be in the target range for achieved concentration, and homogenous.

There were no treatment-related deaths, adverse clinical observations, or necropsy findings during the study. Minor post-dosing observations associated with taste-aversion were seen in many treated animals during the study

Group mean body weight gain was variable with high dose males gaining slightly less weight than controls over the course of the study but this was not dose-related and not statisically significant, therefore it was considered of no biological relevance. There was no clear effect of treatment on mean food intake either before pairing in males and females or in females during gestation.

Mating and fertility data showed no effect of treatment.

After parturition, 2, 1, 1, and 2 females from Groups 1, 2, 3 and 4 respectively showed total litter loss. The distribution of these losses is similar to that seen in previous littering studies conducted at these laboratories in the same strain of rat, and therefore was not considered to be an adverse effect of treatment. There was no effect of treatment on intrauterine data, or on group mean pup data, with the exception of Day 4 mean pup weights in both sexes which were slightly lower than controls but the changes were not statistically significant and within the range of historical control data, therefore it was not considered to be of toxicological relevance.
In conclusion, oral gavage administration of 1000 mg/kg/day Dimethyl Sebacate to male and female rats produced was characterised by slightly decreased body weight gain in males only.

The no-observed-adverse-effect-level (NOAEL) for pup developement was 1000 mg/kg/day.

Short description of key information:
There is a oral combined repeated dose toxicity study with the reproduction/developmental toxicity screening test in rat. This study was performed according to GLP and OECD 422 guideline. No systemic toxicity or effect on fertility or pup development was observed at the limit dose of 1000 mg/kg/day.

Justification for selection of Effect on fertility via oral route:
The combined repeated dose toxicity study with the Reproduction/Developmental toxicity screening test was performed according to OECD 422 guideline and GLP. It is the key study for this endpoint.

Effects on developmental toxicity

Description of key information

The objective of the developmental toxicity study performed according to OECD 414 (Bentz, 2016) was to evaluate the maternal and developmental toxicity of Dimethyl Sebacate in rats. The test item, Dimethyl Sebacate, was administered by gavage, once daily, from Days 6 to 20 p.c. inclusive, to pregnant Sprague-Dawley rats at dosages of 100, 300 and 1000 mg/kg/day.Under the experimental conditions and results of this study: The No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 1000 mg/kg/day in absence of adverse effects in the dams. The NOAEL for embryo-fetal development was considered to be 1000 mg/kg/day in absence of adverse effects in the litters. The test item did not elicit a teratogenic potential.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Remarks:

43037 RSR

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 January 2016 - 10 March 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS - Source: Janvier, le Genest-Saint-Isle, France- Sex: female - Age at study initiation: at the beginning of the treatment period, the animals were 10-11 weeks old - Weight at study initiation: the animals had a mean body weight of 278 g (range: 231 g to 334 g) - Housing: the animals were individually housed in cages (Tecniplast 2154, 940 cm2; polycarbonate with a stainless steel lid) containing autoclaved sawdust - Diet (e.g. ad libitum): free access to SSNIFF R/M-H pelleted maintenance diet - Water (e.g. ad libitum): free access to bottles containing tap water (filtered with a 0.22 µm filter) - Acclimation period: at least 4 days before the beginning of the treatment period. ENVIRONMENTAL CONDITIONS - Temperature (°C): 22 ± 2°C - Humidity (%): 50 ± 20% - Air changes (per hr): about 8 to 15 cycles/hour of filtered, non-recycled air - Photoperiod (hrs dark / hrs light): 12h/12h. IN-LIFE DATES: From: 15 February 2016 To 10 March 2016.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:Solution in the vehicle.Preparation procedure:According to a hompogeneity/stability describing the preparation procedure (stability testing) for a range of concentrations covering the lowest and highest used in this study: the test item was weighed and mixed with the required quantity of vehicle according to CiToxLAB France in house procedures. VEHICLE - Justification for use and choice of vehicle (if other than water): suitable formulation in corn oil - Concentration in vehicle: 20, 60 and 200 mg/mL - Amount of vehicle (if gavage): 5 mL/kg/day.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

High Performance Liquid Chromatography with UV detection (HPLC/UV).Once on formulations used in the first and last weeks of treatment in the study.A sample was taken from control and test item dose formulations and analyzed using the validated method.

Duration of treatment / exposure:

15 days.

Frequency of treatment:

Daily.

Duration of test:

Day 6 to Day 20 p.c.

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:

Groups 1, 3 and 4: 24 females.Group 2: 23 females.

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose-levels were selected in agreement with the Sponsor, and are based on the results of a preliminary prenatal development study where Sprague-Dawley rats received daily the test item by gavage at 100, 300 or 1000 mg/kg/day from Days 6 to 20 p.c. in the same vehicle. There were no adverse findings in this preliminary study at any dose-level. Therefore, the same dose-levels were selected in the present study.- Rationale for animal assignment: computerized stratification procedure.

Maternal examinations:

MORTALITY AND MORBIDITY: Yes - Time schedule: Each animal was checked for mortality and morbidity once a day before the treatment period and at least twice a day during the treatment period, including weekends and public holidays.CLINICAL OBSERVATIONS: Yes - Time schedule: From arrival, each animal was observed once a day as part of the routine examinations.From the start of the treatment period, each animal was observed once a day, at approximately the same time of day, for the recording of clinical signs. BODY WEIGHT: Yes - Time schedule: The body weight of each female was recorded on Days 2, 4, 6, 9, 12, 15, 18 and 21 p.c. FOOD CONSUMPTION: Yes - Time schedule: The quantity of food consumed by each female was recorded for the following intervals: Days 2-4, 4-6, 6-9, 9-12, 12-15, 15-18 and 18-21 p.c. POST-MORTEM EXAMINATIONS: Yes - Sacrifice on gestation Day 21 - Organs examined: principal thoracic and abdominal organs and the weight of the gravid uterus. OTHER:

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes Examinations included:- 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: approximately half of the fetuses in each litter- Skeletal examinations: approximately half of the fetuses in each litter- Head examinations: approximately half of the fetuses in each litter- Other: number dead and live, body weight, sex.

Statistics:

Data were compared by one-way analysis of variance and Dunnett test (mean values being considered as normally distributed and variances being considered as homogeneous) or by Fisher exact probability test (proportions).

Indices:

% Pre-implantation loss: 100 * (Number of corpora lutea - Number of implantations) / Number of corpora lutea% Post-implantation loss: 100 * (Number of implantations sites - Number of live fetuses) / Number of implantation sites

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

At 1000 mg/kg/day, ptyalism, observed in 21/22 pregnant females for 2 days to the entire treatment period, was considered to be related to the test item treatment and of minor toxicological significance. At 100 and 300 mg/kg/day, there were no test item-related clinical signs. Clinical signs recorded at these dose-levels (reflux at dosing, ptyalism and/or reddish vaginal discharge) were noted in comparable incidences with controls.
The summary table is enclosed to this RSS in table 1 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Mortality:

no mortality observed

Description (incidence):

There were no premature deaths.
The summary table is enclosed to this RSS in table 1 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no effects on mean body weight and mean body weight change at any dose-levels.
The summary tables are enclosed to this RSS in tables 2, 3 and 4 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no effects on mean food consumption at any dose-levels.The high food consumption noted for one control female from Days 12 to 15 p.c. (about three times more than the other control females at the same period) was considered to be an aberrant value.
The summary table is enclosed to this RSS in table 5 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

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:

effects observed, non-treatment-related

Description (incidence and severity):

There were no necropsy findings considered to be test item treatment-related (no dose-relationship or isolated finding).
The summary table is enclosed to this RSS in table 6 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Number of abortions:

no effects observed

Description (incidence and severity):

The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

When compared with controls, higher mean pre-implantation losses noted at 100 and 1000 mg/kg/day were not ascribed to the test item treatment as treatment started after implantation (implantation completed in rats on Day 6 p.c.). Slightly higher mean post-implantation loss at 1000 mg/kg/day was considered to be incidental (no statistical significance, increase due to 2/22 dams which also had a high pre-implantation loss and thus less implantation sites).
The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Early or late resorptions:

no effects observed

Description (incidence and severity):

The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Dead fetuses:

no effects observed

Description (incidence and severity):

The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

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): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Changes in number of pregnant:

not examined

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

clinical signs

Remarks on result:

other: no relevant effect observed at the top dose

Remarks:

no relevant effect observed at the top dose

Abnormalities:

not specified

Fetal body weight changes:

no effects observed

Description (incidence and severity):

There were no test item-related effects on mean fetal body weight.
The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Changes in sex ratio:

no effects observed

Description (incidence and severity):

There were no test item-related effects on mean sex ratio.
The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

The summary table is enclosed to this RSS in table 7 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Changes in postnatal survival:

not examined

External malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no external variations.The only fetal external malformation was an anasarca (generalized edema) in one litter at 100 mg/kg/day. In absence of dose-relationship and as this finding was in an isolated incidence, it was not attributed to the test item treatment.
The summary tables are enclosed to this RSS in tables 8 and 9 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

Cartilages of unossified or incomplete/bipartite ossification of bones were present, with the following exceptions:- a bipartite centrum of one thoracic vertebra in one litter of each group treated at 0 or 1000 mg/kg/day, - a 13th rib in one litter at 300 mg/kg/day (leading to a short rib).These findings being isolated and/or also noted in a control litter, they were not attributed to the test item treatment.At 1000 mg/kg/day, there were higher incidences of litters with fetuses having dumbbell ossification of thoracic vertebra(e) centrum and/or unossified distal phalanx of hindpaw and/or forepaw. As the variations from controls were generally slight and not statistically significant, and in absence of effects on mean fetal body weight, these tendencies were considered to be of no toxicological significance.The other skeletal variations were not considered to be test item treatment-related (they were noted in comparable or lower incidence than in controls or historical control data, were noted in isolated occurrence and/or were not dose-related).There were no fetal skeletal malformations in test item-treated groups.
The summary tables are enclosed to this RSS in tables 12, 13 and 14 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no fetal soft tissue variations considered to be test item treatment-related. The variations noted in test item groups were of comparable incidence of controls (of the current study or of recent similar studies), isolated and/or not dose-related.These malformations were noted in isolated fetuses, at different dose-levels and on different organs. Right sided aortic arch has already been observed spontaneously in our laboratory (in a control fetus of Historical control data of 2012) and the variation: dilatation of one lateral cerebral ventricle was noted in one control fetus in this study remaining the malformation noted at 1000 mg/kg/day. Therefore, a relationship of these malformations with the test item treatment was considered unlikely.
The summary tables are enclosed to this RSS in tables 10 and 11 in the following attached file: 106-79-6 OECD 414 rat_summary tables.pdf.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

The test item, Dimethyl Sebacate, was administered by gavage, once daily, from Days 6 to 20 p.c. inclusive, to pregnant Sprague-Dawley rats at dosages of 100, 300 and 1000 mg/kg/day.Under the experimental conditions and results of this study:- the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 1000 mg/kg/day in absence of adverse effects in the dams, - the NOAEL for embryo-fetal development was considered to be 1000 mg/kg/day in absence of adverse effects in the litters,- the test item did not elicit a teratogenic potential.

Executive summary:

The objective of this GLP study was to evaluate the potential toxic effectsof the test item, Dimethyl Sebacate, on the pregnant female and on embryonic and fetal development, following daily oral administration (gavage) to pregnant female rats from implantation to the day before scheduled hysterectomy [Day 6 to Day 20 post-coitum (p.c.) inclusive].

 

Methods

Three groups of 23 (one female excluded before study beginning) or 24 time-mated female Sprague-Dawley rats received the test item, Dimethyl Sebacate, at 100, 300 or 1000 mg/kg/day by oral route (gavage) once daily from Days 6 to 20 p.c. A constant dosage-volume of 5 mL/kg/day was used. Another group of 24 rats received the vehicle alone (corn oil) under the same experimental conditions, and acted as a control group.

 

Formulation concentrations were checked in the first and last weeks of treatment in the study.

 

The animals were checked daily for mortality and clinical signs. Body weight and food consumption were recorded every 2 to 3 days. On Day 21 p.c., females were sacrificed and submitted to a macroscopic post-mortem examination. Hysterectomy was performed and the numbers of corpora lutea, uterine scars, implantations, early and late resorptions, and live and dead fetuses were recorded. The fetuses were sexed, weighed and examined for external, soft tissues and skeletal (cartilages + bones) abnormalities.

 

Results

The test item concentrations analyzed were within an acceptable range of variations when compared to the nominal values and no test item was observed in the control dose formulations.

 

At 1000 mg/kg/day, ptyalism, observed in 21/22 pregnant females for 2 days up to the entire treatment period, was considered to be related to the test item treatment and of minor toxicological significance.

 

There were no premature deaths and notest item-relatedeffects onpregnancy status,mean body weight, mean body weight gain, mean food consumption, mean gravid uterus weight, mean carcass weight, mean net body weight change and mean hysterectomy data. There were no necropsy findings considered to be test item treatment-related.

 

In litters, there were no test item-related effects on mean fetal body weight or mean sex ratio.

There were no fetal external variations or skeletal malformations, and no test item-related external malformations or soft tissue variations/malformations or effects on cartilages. There were no toxicologically significant skeletal variations.

 

Conclusion

The test item, Dimethyl Sebacate, was administered by gavage, once daily, from Days 6 to 20 p.c.inclusive, to pregnant Sprague-Dawley rats at dosages of 100, 300 and 1000 mg/kg/day.

 

Under the experimental conditions and results of this study:

.         the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 1000 mg/kg/day in absence of adverse effects in the dams,

.         the NOAEL for embryo-fetal development was considered to be 1000 mg/kg/day in absence of adverse effects in the litters,

.         the test item did not elicit a teratogenic potential.

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:

Good quality

Justification for classification or non-classification

According to the available data, Dimethyl Sebacate does not warrant classification for reproductive and developmental toxicity according to Regulation (EC) No 1272-2008 and Annex VI of Commission Directive 2001/59/EC