Tri-C18-22 (even numbered)-alkyl 2-hydroxypropane-1,2,3-tricarboxylate

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Published study conducted to GLP and a recognised guideline

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: ICH Harmonised Tripartite Guidelines for Detection of Toxicity to Reproduction for Medicinal Products

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Behenyl alcohol

IUPAC Name:

Behenyl alcohol

Details on test material:

Behenyl alcohol is a saturated 22-carbon, long-chain aliphatic alcohol. The composition and stability of behenyl alcohol was documented throughout the study, no further information is provided.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

The test animals were sexually mature female New Zealand White rabbits, approximately 18 to 26 weeks of age, obtained from Froxfield SPF Rabbits Ltd. (England). Approximately 2 weeks prior to arrival at the test facility, oestrus was synchronised by the supplier by an intravenous injection of 25 IU lutenizing hormone. Upon arrival, rabbits were allowed to acclimatise for a minimum of 1 week prior to the initiation of dosing. The rabbits weighed 3.29-4.98 kg at study initiation. The rabbits were housed individually in suspended stainless-steel cages equipped with an undertray lined with absorbent paper. The housing room was maintained at 18°C and 55% relative humidity with a 12 hour light-dark cycle. Rabbits were provided with a standard rabbit diet (Special Diet Services Ltd., England) and tap water, ad libitum.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 1% w/w Tween 80

Details on exposure:

The test substance was administered by gavage in 1% w/w Tween 80 at 0, 125, 500 or 2000 mg/kg bw/d. The preparation was administered at volume-dosages of 10, 0.625, 2.5 or 10 ml/kg bw/d corresponding to doses of 0. 125, 500 and 2000 mg/kg bw/d, respectively. The volume administered to each animal was calculated from bodyweights measured on the same day that dosing took place.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No information available.

Details on mating procedure:

Females were mated with New Zealand White males of proven fertility. Following insemination, each female was injected i.v. with 25 IU of lutenizing hormone to ensure successful ovulation. The day of insemination was designated as Day 0 of gestation. All animals were examined on Day 6 of gestation and determined to be suitable for use in the study.

Duration of treatment / exposure:

16 days - Day 6 to Day 19 of gestation

Frequency of treatment:

Once daily between Day 6 to 19 of gestation

Duration of test:

16 days - Day 6 to Day 19 of gestation

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 125, 500 and 2000 mg/kg bw/d
Basis:
nominal conc.

No. of animals per sex per dose:

22 females/group

Control animals:

yes, concurrent vehicle

Details on study design:

The dose levels chosen for the study were selected based on a previous range-finding study providing a maximum dose of 2000 mg/kg bw/d. Following mating, females were randomly allocated to the 4 treatment groups in order of mating to evenly distribute the mated females among the groups.

Examinations

Maternal examinations:

Cage-side observations were made daily for signs of toxicity and moribundity. Food consumption was recorded during the following time points: Day 1 to 5, 6 to 12, 13 to 19, 20 to 23 and 24 to 28, inclusive. Bodyweight gains and water consumption were measured daily. Any animal that delivered prematurely was killed by an i.v. injection of pentobarbitone sodium B Vet C. on the same day that the premature delivery was detected.
Following euthanasia on Day 28 of gestation, the rabbits were examined macroscopically for evidence of disease and signs of toxicity.

Ovaries and uterine content:

The rabbits were killed on Day 28 of gestation by an i.v. injection of pentobarbitone sodium and the uterine contents examined. The number of corpora lutea were recorded, and the reproductive tract, including the ovaries was dissected out. For each animal, the number of pre- and post-implantation sites, early and late resorptions, and viable foetuses, as well as the distribution of foetuses in the uterine horn were examined. The uterus of any female presumed to be non-pregnant was stained using a 10% aq (v/v) ammonium sulphide solution and examined for implantation sites.

Fetal examinations:

Each foetus was weighed and examined externally. The foetuses were killed by an s.c. injection of pentobarbitone sodium, and uniquely identified within the litter with respect to their uterine position. Placental weights were recorded and examined macroscopically for any abnormalities. The neck, thoracic and abdominal cavities from each foetus were dissected and the contents examined microscopically. One third of the foetuses were decapitated and the heads fixed in Bouin's fluid for examination using a modification of the Wilson free-hand serial sectioning technique. All foetuses were subsequently eviscerated, fixed in methylated spirit and processed and stained with Alizarin Red for skeletal examination.

Statistics:

One way ANOVA was performed on bodyweights, bodyweight changes, and food and water consumption. Organ weights were evaluated by Dunnett's or Behren's-Fisher's tests. Nested ANOVA and weighed t-test were conducted on foetal and placental weights.

Indices:

Not determined.

Historical control data:

No information available.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
All females survived until study termination. There were no clinical signs of toxicity, with the exception of pale faeces observed in the majority of 2000 mg/kg bw/d females. There were no effects of treatment on bodyweight gain and food and water consumption. There were no treatment related findings at necropsy. A total of 3 females, treated with 0, 125 and 500 mg/kg bw/d, respectively, were observed to have total litter loss at necropsy. The uterus of each of these females revealed early resorptions.

Effect levels (maternal animals)

1

Dose descriptor:

NOAEL

Effect level:

2 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

2

Dose descriptor:

NOAEL

Effect level:

2 000 mg/kg bw/day (nominal)

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:
There were no effects of treatment on the number of corpora lutea, pre- and post-implantation sites, early and late resorptions, and viable foetuses. Foetal and placental weights were not affected by treatment. Sex ratios were comparable across groups. No macroscopic, skeletal or visceral variations were observed that were not comparable to historical control values previously reported for the laboratory.

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Parameter	Dose (mg behenyl alcohol/kg bw/d)
	0	125	500	2000
No. pregnant animals	20	19	19	20
Premature delivery and total litter loss (%)	10.0	5.3	5.3	0.0
Corpora lutea count	12.8 ± 3.1	12.9 ± 2.2	12.6 ± 3.0	12.2 ± 3.9
Implantations	11.4 ± 3.9	11.1 ± 2.6	11.0 ± 3.3	10.6 ± 4.3
Viable young
Male	4.6 ± 2.6	4.8 ± 1.5	3.8 ± 1.5	4.7 ± 2.2
Female	5.5 ± 2.4	4.9 ± 1.7	5.5 ± 2.1	4.3 ± 2.5
Total	10.1 ± 3.7	9.8 ± 2.1	9.3 ± 2.6^a	9.0 ± 3.8
Resorptions
Early	0.4 ± 0.6	0.3 ± 0.5	0.4 ± 0.6	0.7 ± 0.8
Late	1.0 ± 1.0	1.1 ± 1.0	1.2 ± 1.1	0.9 ± 0.9
Total	1.4 ± 1.2	1.3 ± 1.2	1.7 ± 1.3	1.6 ± 1.2
Implantation loss (%)
Pre	10.4	14.2	13.9	13.5
Post	12.1	12.0	15.2	14.7

Data represent mean ± standard deviation

^aIncludes one foetus not sexed at necropsy

Applicant's summary and conclusion

Conclusions:

The NOAEL is considered to be 2000 mg/kg bw/d for both maternal toxicity and developmental toxicity.

Executive summary:

A developmental toxicity study was conducted with behenyl alcohol. The test substance was administered to mated female New Zealand White rabbits daily by gavage at doses of 0, 125, 500 and 2000 mg/kg bw/d on Day 6 through to Day 19 of gestation. The dams were sacrificed at Day 28 of gestation for examination of the uterine contents.

There were no maternal deaths, and no clinical signs of toxicity, with the exception of pale faeces observed in the majority of 2000 mg/kg bw/d females. There were no effects of treatment on bodyweight gain and food and water consumption. There were no treatment related findings at necropsy. A total of 3 females, treated with 0, 125 and 500 mg/kg bw/d, respectively, were observed to have total litter loss at necropsy. The uterus of each of these females revealed early resorptions. There were no effects of treatment on the number of corpora lutea, pre- and post-implantation sites, early and late resorptions, and viable foetuses. Foetal and placental weights were not affected by treatment. Foetal sex ratios were comparable across groups. No macroscopic, skeletal or visceral variations were observed.

There was no evidence that behenyl alcohol is teratogenic or embryotoxic. The NOAEL is considered to be 2000 mg/kg bw/d for both maternal toxicity and developmental toxicity.