Reaction mass of N,N’-ethane-1,2-diylbis(12-hydroxyoctadecan-1-amide), Octadecanamide, 12-hydroxy-N-[2-[(1-oxooctadecyl)amino]ethyl]- and Octadecanoic acid, 12-hydroxy-, 1-hexyl-12-[[2-[(12-hydroxy-1-oxooctadecyl)amino]ethyl]amino]-12-oxododecyl ester

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 August 2017 - 16 October 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

22 January 2001

Deviations:

yes

Remarks:

See for details Any Other information on Materials and Methods

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Octadecanoic acid, 12-hydroxy-, reaction products with ethylenediamine

EC Number:

309-629-8

EC Name:

Octadecanoic acid, 12-hydroxy-, reaction products with ethylenediamine

Cas Number:

100545-48-0

IUPAC Name:

309-629-8

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: breeder: Janvier, le Genest-Saint-Isle, France
- Age at the beginning of the treatment period: the animals were 10-11 weeks old
- Mean body weight at the beginning of the treatment period: the animals had a mean body weight of 296 g (range: 245 g to 358 g)
- Fasting period before study: no
- Housing: the animals were individually housed in polycarbonate cages (Tecniplast 2154, 940 cm2) with stainless steel lids and containing autoclaved sawdust
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: for a period of 4 or 5 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): 12 h/12 h.

IN-LIFE DATES: 11 September 2017 to 16 October 2017.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION
The test item was administered as a suspension in the vehicle. The test item was ground to a fine powder, using a mortar and pestle, and then mixed with the required quantity of vehicle.
The test item dose formulations were prepared daily and were delivered to the study room at room temperature and protected from light.
This preparation process was validated for a range of concentrations covering the lowest and highest concentrations used in this study.
.

VEHICLE
- Justification for use and choice of vehicle (if other than water): homogeneous suspensions were obtained with corn oil as a vehicle
- Concentration in vehicle: 20, 60 and 200 mg/ml
- Amount of vehicle (if gavage): 5 ml/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Type of method: High Performance Liquid Chromatography with tandem Mass Spectrometry detection (LC/MS MS)
Test item concentrations: the test item concentrations in the administered dose formulations analyzed on Study Day 5, in study Week 4 and instudy week 5 of the treatment period, were within an acceptable range of variation (-12.1% to +6.0%) when compared to the nominal values (± 15% required).
Homogeneity: The dose formulations containing the test item and prepared at 10 mg/mL and 200 mg/mL in corn oil were found to be homogeneous at room temperature and protected from light.
Stability: UVCB, dose formulation prepared daily

Details on mating procedure:

Impregnation procedure: purchased time-mated females
Females arrived on Day 1 or 2 post-coitum

Duration of treatment / exposure:

Days 6 to 20 post-coitum

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

24 females

Control animals:

yes, concurrent vehicle

Details on study design:

- Rationale for dose selection:
The dose levels were selected on the basis of the results of a OECD 421 study (reproduction/developmental toxicity screening study in the Sprague-Dawley rat by oral gavage administration), where the NOAEL was set at 1000 mg/kg/day. During the first week of treatment, test item treatment-related reductions in body weight gain were apparent in all groups of treated females but the animals quickly recovered to show normal growth patterns through the rest of the study.
Therefore, 1000 mg/kg/day was selected as the high-dose level. The low-dose and mid dose were selected using a ratio representing approximately a 3-fold interval (i.e. 100 and 300 mg/kg/day).

- Rationale for animal assignment: computerized stratification procedure.

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: 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.

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 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 post-coitum

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
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 post-coitum

WATER CONSUMPTION: No

POST-MORTEM MACROSCOPIC EXAMINATION: Yes
- Sacrifice on Day 21 post-coitum.
- Examined: principal thoracic and abdominal organs.

Ovaries and uterine content:

The ovaries and uterine content were examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
The weight of the gravid uterus was recorded for each pregnant female (with at least one live fetus) at hysterectomy.
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
- Number and distribution of dead and live fetuses,
- Number and distribution of uterine scars (uterine implantation without implant)

The following classification was used to record:
. uterine scar: uterine implantation without implant,
. early resorption: evidence of implant without recognizable embryo,
. late resorption: dead embryo or fetus with degenerative changes,
. dead fetus: dead fetus with no degenerative changes.

Fetal examinations:

- External examinations: Yes: all fetuses per litter
- Soft tissue examinations: Yes: half fetuses per litter
- Skeletal examinations: Yes: half fetuses per litter
- Head examinations: Yes: half fetuses per litter
- Other: number of dead and alive fetuses , body weight and sex of fetuses

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’s exact probability test (proportions).

Indices:

The following parameters were calculated:
For each pregnant female:
- Body weight change for different intervals
- Net body weight (presented as carcass weight) = Body weight on Day 21 post-coitum - gravid uterine weight
- Net body weight change = Body weight on Day 21 post-coitum - body weight on Day 6 post-coitum - gravid uterine weight%

For each litter:
- Total number of resorptions = Sum of uterine scars + early resorptions + late resorptions
- Total number of dead fetuses = Sum of dead fetuses
- % of dead fetuses per litter = (Total number of dead fetuses / Number of implantation sites) x 100
- Total number of live fetuses = Sum of live male + live female fetuses
- % of live fetuses per litter = (Total number of live fetuses / Number of implantation sites) x 100
- % of pre-implantation loss = (Number of corpora lutea - Number of implantations / Number of corpora lutea) x 100
- % of post-implantation loss = (Number of implantation sites - Number of live fetuses / Number of implantation sites) x 100
- Average fetal body weight= Sum of individual fetal weights / Number of live fetuses

For each group:
- % of pre-implantation loss relative to the number of corpora lutea (mean of pre-implantation loss per litter)
- % of live fetuses and % of dead fetuses (relative to total number of fetuses)
- Mean % of male fetuses per litte
- Mean and standard deviations and % relative to the number of implantation sites: resorptions plus, uterine scars, uterine scars, early resorptions, late
resorptions,
- % of post-implantation loss relative to the number of implantation sites (mean of post-implantation loss per litter), % of dams affected.

Historical control data:

See attached document

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, non-treatment-related

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:

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

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Description (incidence and severity):

There were no effects on mean carcass weight, mean gravid uterus weight, or mean net body weight change.

Details on results:

CLINICAL SIGNS AND MORTALITY
There was no mortality observed during the study.
There were no test item-related clinical signs. Clinical signs seen at 100 and 300 mg/kg/day (cutaneous lesion, mass on mammary gland and scabs) were without dose-relationship and observed in isolated animals (3 and 1 animals at 100 and 300 mg/kg bw respectively).

BODY WEIGHT AND WEIGHT CHANGE (cf table 7.8.2/1 in chapter any other information on results)
There were no effects on mean body weight and mean body weight change.

FOOD CONSUMPTION
There were no treatment related changes on food consumption.

GROSS PATHOLOGY
There were no test item-related necropsy findings.
The mammary gland mass seen at 100 mg/kg/day (female L20663) and the mass on one placenta at 300 mg/kg/day (female L20676) were considered to be incidental in absence of dose-relationship and as it was noted in isolated animals.

OTHER EFFECTS (cf table 7.8.2/2 in chapter any other information on results)
There were no effects on mean carcass weight, mean gravid uterus weight, or mean net body weight change.

Maternal developmental toxicity

Pre- and post-implantation loss:

effects observed, non-treatment-related

Total litter losses by resorption:

effects observed, non-treatment-related

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

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): not examined

Changes in number of pregnant:

no effects observed

Details on maternal toxic effects:

PRE AND POST IMPLANTATION LOSS (cf table 7.8.2/3 in chapter any other information on results)
There were no test item-related effects on pre and post implantation losses at any dose-level.
At 1000 mg/kg/day, the 100% of post-implantation loss noted in female L20708 induced an increase of the mean percentage of post-implantation loss in this dose group (11.7% compared to 6.4% in the control group). This was considered to be incidental and due to the fact that female L20708 had only one corpora lutea. When excluding L20708 data, the mean percentage of post-implantation loss in the 1000 mg/kg bw/day group was 7.6%

TOTAL LITTER LOSSES BY RESORPTION (cf table 7.8.2/3 in chapter any other information on results)
At 1000 mg/kg/day, a total litter loss occured in female L20708 but this was considered incidental as this female had only one implantation site with one uterine scar. No other total litter losses were observed whatever the dose levels.

EARLY OR LATES RESORPTIONS (cf table 7.8.2/3 in chapter any other information on results)
There were no test item-related effects on early or late resorptions.

DEAD FETUSES (cf table 7.8.2/3 in chapter any other information on results)
There were no dead fetuses.

CHANGES IN NUMBER OF PREGNANT (cf table 7.8.2/3 in chapter any other information on results)
At hysterectomy on Day 21 post-coitum., there were 24/24, 24/24, 23/24 and 23/24 pregnant dams in the groups treated at 0, 100, 300 or 1000 mg/kg/day, respectively.

Effect levels (maternal animals)

Key result

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified

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:

no effects observed

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

See tables 5 and 6.
The visceral variations were considered to be incidental as there was no dose-relationship, they were within Historical Control Data, differences from controls or Historical control maximum were slight, and/or they were noted in single fetuses.
The malformations observed at 100 and 1000 mg/kg/day were considered to be unlikely related to the test item treatment as they were noted in isolated fetuses and/or without dose-relationship.

Details on embryotoxic / teratogenic effects:

FETAL BODY WEIGHT (cf table 7.8.2/4 in chapter any other information on results)
There were no effects on mean fetal body weight.

REDUCTION IN NUMBER OF LIVE FETUSES (cf table 7.8.2/3 in chapter any other information on results)
There were no reduction in number of live fetuses compared to controls.

CHANGE IN SEX RATIO (cf table 7.8.2/4 in chapter any other information on results)
There were no effects on mean percentage of male fetuses.

CHANGE IN LITTER SIZE and WEIGHTS (cf tables 7.8.2/3 and 7.8.2/4 in chapter any other information on results)
There were no changes in litter size and weights compared to controls

EXTERNAL MALFORMATIONS:
There were no external malformations in test item treated groups.

SKELETAL MALFORMATIONS (cf tables 7.8.2/5 and 7.8.2/6 in chapter any other information on results and pdf file in attached background material)
There were no fetal skeletal malformations in test item-treated groups.
Fetal skeletal variations included extra sternebral ossification sites, misshapen sternebrae and unossified distal phalanx of the forepaws were observed at litter incidence higher than controls. None of them were attributed to treatment as their incidences were not dose-related, isolated and the fetal incidences were lower than or close to those of controls.

VISCERAL MALFORMATIONS (cf tables 7.8.2/7 in chapter any other information on results and historical data in attached background material)
In the 100 mg/kg bw/day group, kidneys were absent in 0.7% of the fetuses. In the 1000 mg/kg bw/day group, aortic arch was absent in 0.8% of the fetuses. These malformations were considered to be unlikely related to the test item treatment as they were noted in isolated fetuses without dose-relationship.

Visceral variations included dilated renal pelvis, dilated ureters, lack or short (innominate) arteries and reddish foci in thymus. They were all considered to be incidental:
Dilated renal pelvis were not dose-related and in within fetal incidence of controls. Dilated ureters were not dose-related and within fetal and litter incidences of historical control data. At 1000 mg/kg bw/day, lack of (innominate) arteries were slightly above controls but within fetal and litter incidences of historical control data. Short (innominate) arteries were similar to fetal and litter incidences of controls. Reddish foci in thymus were not dose-related and observed in a single fetus of the 100 and 300 mg/kg bw/day groups.

Effect levels (fetuses)

Key result

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

Table 7.8.2/1: Mean Body weight and mean body weight change (g) in pregnant females with live concepti:

Dose level (mg/kg/day)	0	100	300	1000
Body weight
Day 6 post-coitum	299	295	295	297
		-1	-1	-1
Day 21 post-coitum	450	442	443	453
		-2	-2	+1
Body weight change
Days 6 - 21 post-coitum	+151	+147	+147	+156

in italic: differences from controls (%)

Table 7.8.2/2: Mean gravid uterus, mean carcass weights and mean net body weight change (g) in pregnant females with live concepti:

 

Dose level (mg/kg/day)	0	100	300	1000
Gravid uterus weight	99.6	98.9	101.8	105.0
Carcass weight^a	350	343	341	348
Net body weight change from Day 6post-coitum	51.5	48.2	45.2	50.8

a: values rounded to three significant digits.

Table 7.8.2/3: Hysterectomy data

 

Dose (mg/kg/day)		0	100	300	1000
Pregnant Females Alive at Term	N	24	24	23	23
With Total Resorptions	N	0	0	0	1*
With all Dead Fetuses	N	0	0	0	0
With Live Fetuses	N	24	24	23	22
Corpora Lutea	TOTAL MEAN SD	344 14.3 2.5	353 14.7 2.4	313 13.6 2.1	320 13.9 3.1
N° per animal
Implantation Sites	TOTAL MEAN SD	314 13.1 2.2	317 13.2 2.4	298 13.0 2.2	303 13.2 3.0
N° per animal
Preimplantation Loss	MEAN % SD	8.4 9.6	10.0 11.2	4.8 5.2	5.0 5.1
Fetuses	TOTAL MEAN SD	295 12.3 2.6	296 12.3 2.3	283 12.3 2.2	279 12.1 3.2
N° per animal
Live Fetuses	TOTAL MEAN SD MEAN% SD	295 12.3 2.6 93.6 9.4	296 12.3 2.3 93.5 5.7	283 12.3 2.2 95.7 11.2	279 12.1 3.2 88.3 21.6
Dead Fetuses	TOTAL MEAN % SD	0 0.0 0.0	0 0.0 0.0	0 0.0 0.0	0 0.0 0.0
Resorptions+Scars	TOTAL     MEAN SD	19 6.1   0.8 1.2	21 6.6   0.9 0.7	15 5.0   0.7 1.9	24 7.9   1.0 1.3
% of implantation sites
N° per animal
Implant Scars	TOTAL     MEAN SD	0 0.0   0.0 0.0	0 0.0   0.0 0.0	0 0.0   0.0 0.0	1 0.3   0.0 0.2
% of implantation sites
N° per animal
Resorptions: early	TOTAL     MEAN SD	18 5.7   0.8 1.2	18 5.7   0.8 0.7	15 5.0   0.7 1.9	20 6.6   0.9 1.4
% of implantation sites
N° per animal
Resorptions: late	TOTAL     MEAN SD	1 0.3   0.0 0.2	3 0.9   0.1 0.3	0 0.0   0.0 0.0	3 1.0   0.1 0.5
% of implantation sites
N° per animal
Postimplantation Loss   Dams Affected	MEAN%  SD N %	6.4  9.4 11 45.8	6.5  5.7 16 66.7	4.3  11.2 7 30.4	11.7 **  21.6 13 56.5

*: female L20708    **: mean of 7.6% when excluding data of female L20708

Table 7.8.2/4: Mean fetal body weight (g) and sex ratio

Dose (mg/kg/day)		0	100	300	1000
Fetal Body Weight (g)/ Litter (N)	N MEAN SD	24 5.76 0.41	24 5.72 0.53	23 5.97 0.46	22 5.93 0.31
Males Fetuses/ Litter (N)	N MEAN SD	24 5.93 0.45	24 5.87 0.56	23 6.15 0.45	22 6.10 0.31
Males Fetuses	MEAN% SD	50.8 14.8	43.7 14.4	49.4 12.3	50.5 16.2

  

Table 7.8.2/5: Total fetal skeletal malformations

Dose (mg/kg/day)		0	100	300	1000
Litters Evaluated	N	24	24	23	22
Fetuses Evaluated	N	154	154	146	146
Fetal Incidence	N %	7 4.5	0 0.0	0 0.0	0 0.0
Litter Incidence	N %	5 20.8	0 0.0	0 0.0	0 0.0
Affected Fetuses /Litter	MEAN% SD	5.3 11.1	0.0 0.0	0.0 0.0	0.0 0.0

 

Table 7.8.2/6: Total fetal skeletal variations

Dose (mg/kg/day)		0	100	300	1000
Litters Evaluated	N	24	24	23	22
Fetuses Evaluated	N	154	154	146	146
Fetal Incidence	N %	113 73.4	113 73.4	94 64.4	99 67.8
Litter Incidence	N %	20 83.3	23 95.8	19 82.6	22 100
Affected Fetuses /Litter	MEAN% SD	72.9 35.8	72.8 29.3	62.1 38.6	67.5 25.2

 

 

Table 7.8.2/7: Fetal soft tissue malformations

Dose (mg/kg/day)		0	100	300	1000
Litters Evaluated	N	24	24	23	22
Fetuses Evaluated	N	141	142	137	133
MOUTH, JAW, PALATE
Litter Incidence	N	1	0	0	0
Fetal Incidence	N	1	0	0	0
CLEFT PALATE
Fetal Incidence	N %	1 0.7	0 0.0	0 0.0	0 0.0
Litter Incidence	N %	1 4.2	0 0.0	0 0.0	0 0.0
Affected Fetuses /Litter	MEAN % SD	0.8 4.1	0.0 0.0	0.0 0.0	0.0 0.0
KIDNEYS
Litter Incidence	N	0	1	0	0
Fetal Incidence	N	0	1	0	0
ABSENT KIDNEY
Fetal Incidence	N %	0 0.0	1 0.7	0 0.0	0 0.0
Litter Incidence	N %	0 0.0	1 4.2	0 0.0	0 0.0
Affected Fetuses /Litter	MEAN % SD	0.0 0.0	0.7 3.4	0.0 0.0	0.0 0.0
VESSELS
Litter Incidence	N	0	0	0	1
Fetal Incidence	N	0	0	0	1
ABSENT AORTIC ARCH
Fetal Incidence	N %	0 0.0	0 0.0	0 0.0	1 0.8
Litter Incidence	N %	0 0.0	0 0.0	0 0.0	1 4.5
Affected Fetuses /Litter	MEAN % SD	0.0 0.0	0.0 0.0	0.0 0.0	0.6 3.0
TOTAL FETAL SOFT TISSUE MALFORMATIONS
Fetal Incidence	N %	1 0.7	1 0.7	0 0	1 0.8
Litter Incidence	N %	1 4.2	1 4.2	0 0.0	1 4.5
Affected Fetuses /Litter	MEAN % SD	0.8 4.1	0.7 3.4	0.0 0.0	0.6 3.0

Table 7.8.2/8: Fetal soft tissue variations

Dose (mg/kg/day)		0	100	300	1000
Litters Evaluated	N	24	24	23	22
Fetuses Evaluated	N	141	142	137	133
URETER
Litter Incidence	N	5	5	7	6
Fetal Incidence	N	7	6	9	9
DILATED URETER
Fetal Incidence	N %	7 5.0	6 4.2	9 6.6	9 6.8
Litter Incidence	N %	5 20.8	5 20.8	7 30.4	6 27.3
Affected Fetuses /Litter	MEAN % SD	4.9 10.8	4.7 11.3	6.1 10.2	6.5 12.8
KIDNEYS
Litter Incidence	N	2	3	2	2
Fetal Incidence	N	3	3	2	2
DILATED RENAL PELVIS
Fetal Incidence	N %	3 2.1	3 2.1	2 1.5	2 1.5
Litter Incidence	N %	2 8.3	3 12.5	2 8.7	2 9.1
Affected Fetuses /Litter	MEAN % SD	2.2 8.4	2.2 6.3	1.3 4.5	1.5 4.9
VESSELS
Litter Incidence	N	3	2	2	4
Fetal Incidence	N	4	2	2	6
ABSENT INNOMINATE ARTERY
Fetal Incidence	N %	3 2.1	2 1.4	2 1.5	5 3.8
Litter Incidence	N %	3 12.5	2 8.3	2 8.7	4 18.2
Affected Fetuses /Litter	MEAN % SD	1.9 5.1	1.3 4.4	1.2 4.1	3.6 8.5
SHORT INNOMINATE ARTERY
Fetal Incidence	N %	1 0.7	0 0.0	0 0.0	1 0.8
Litter Incidence	N %	1 4.2	0 0.0	0 0.0	1 4.5
Affected Fetuses /Litter	MEAN % SD	0.6 2.9	0.0 0.0	0.0 0.0	0.8 3.6
THYMUS
Litter Incidence	N	0	1	1	0
Fetal Incidence	N	0	1	1	0
REDDISH FOCUS
Fetal Incidence	N %	0 0.0	1 0.7	1 0.7	0 0.0
Litter Incidence	N %	0 0.0	1 4.2	1 4.3	0 0.0
Affected Fetuses /Litter	MEAN % SD	0.0 0.0	0.6 2.6	0.7 3.5	0.0 0.0
TOTAL FETAL SOFT TISSUE MALFORMATIONS
Fetal Incidence	N %	11 7.8	7 4.9	12 8.8	14 10.5
Litter Incidence	N %	7 29.2	5 20.8	8 34.8	8 36.4
Affected Fetuses /Litter	MEAN % SD	7.4 13.3	5.4 12.5	8.0 12.7	10.2 21.9

 

 

Applicant's summary and conclusion

Conclusions:

The No Observed Effect Level (NOEL) for maternal parameters and for embryo-fetal development was considered to be 1000 mg/kg/day (highest dose employed) in absence of substance-related effects in the study.

Executive summary:

In a prenatal development toxicity study performed according to OECD 414 and in compliance with Good Laboratory Practice, the objective was to evaluate the potential toxic effects of the test substance on the pregnant female rat and on embryonic and fetal development, following oral administration (gavage) from Day 6 to Day 20 post-coitum, inclusive. 

Three groups of 24 time-mated female rats received the test substance at doses of 100, 300 or 1000 mg/kg bw /day from Day 6 to 20 post-coitum. A similarly constituted Control group received the vehicle, corn oil, at the same dose volume throughout the same period. Test substance concentration was checked four times in formulations given to the animals

Animals were killed on Day 21 of gestation for reproductive assessment and fetal examination.

Clinical observations, bodyweight and food consumption were monitored. Adult females were examined macroscopically at necropsy on Day 21 of gestation and the numbers of corpora lutea, implantations, early and late resorptions, and live and dead fetuses were recorded. All fetuses were weighed, sexed and examined for external, soft tissue and skeletal abnormalities.

The substance concentrations in the analyzed dose formulations were within an acceptable range of variation when compared to the nominal values (± 15% required), with the exception of formulations administered on Day 6 post coitum (Study Day 1) for 4/24 females per group (between -19.8% and -16.6%). This marginal underdosing was considered to have no impact on the study as there were no major differences in results between these females and the other of the same groups at the end of the study.

At hysterectomy on Day 21 post-coitum., there were 24/24, 24/24, 23/24 and 22/24 pregnant dams with live fetuses in the groups treated at 0,100, 300 or 1000 mg/kg/day,respectively.

There were no test item treatment-related effects in the dams in terms of mortality, clinical signs, necropsy findings, body weight, food consumption, carcass weight, gravid uterus weight, net body weight change, or hysterectomy data. There were no substance treatment-related effects in the litters in terms of sex ratio, fetal body weight, external, visceral and skeletal variations or malformations or cartilage findings.

The No Observed Effect Level (NOEL) for maternal parameters and for embryo-fetal development was considered to be 1000 mg/kg/day (highest dose employed) in absence of treatment related effects in the study.