Docosyl methacrylate

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Principles of method if other than guideline:

A fertility and reproduction study in rats administered a maximum dose of 1000 mg/kg bw/day Docosan-1-ol was conducted.

GLP compliance:

yes

Remarks:

Huntingdon Life Sciences Ltd (Suffolk, England)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Docosan-1-ol

EC Number:

211-546-6

EC Name:

Docosan-1-ol

Cas Number:

661-19-8

Molecular formula:

C22H46O

IUPAC Name:

docosan-1-ol

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited (Margate,Kent, England)
- Age at study initiation: (P) males: 6 to 7 weeks; females: 10 to 11 weeks
- Weight at study initiation: (P) Males:193 and 240 g; Females: 208 and 262 g;
- Housing: TR18 stainless-steel cage (Modular Systems and Developments Company Limited, Hereford, England); during mating period, 1 male and 1 female were
housed in a RB3-modified high-grade polypropylene cage with stainless-steel mesh lids and floors (North Kent Plastic Cages Limited, Erith, Kent, England).
- Diet: ad libitum, expanded rodent diet (Special Diets Services Ltd., Witham, Essex, England) containing no added antibiotic, or other chemotherapeutic
or prophylactic agent
- Water: ad libitum, tap water
- Acclimation period: 6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18°C
- Humidity (%): 55%
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 1% w/w Tween 80

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Whe required amount of behenyl alcohol was weighed into a glass container and heated (approximately 80°C) until molten using an electric mantle.
An appropriate volume of vehicle (1% Tween 80) was heated in a water bath to at least 75°C and then combined with the molten behenyl alcohol under continuous magnetic stirring, to a concentration of 20% behenyl alcohol. The resulting suspension was slowly cooled, with homogenization to a temperature of below 60°C, and then further cooled in a water bath to a temperature of 30°C. The 20% behenyl alcohol suspension was prepared weekly.
The lower concentrations were prepared on the day of use by dilution of the 20% suspension with 1% w/w aqueous Tween 80.

VEHICLE
- aqueous Tween 80
- Amount of vehicle (if gavage): 1%

Animals received the test material or vehicle control formulations by gavage, at volume-dosage of 5 mL/kg bw, using an 8 or 10 choke rubber catheter.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The composition and stability of behenyl alcohol were documented throughout the study.

Duration of treatment / exposure:

Males were treated with Behenyl alcohol daily for 71 days prior to mating, during mating, and until termination. Females were treated with the test
substance for 15 days prior to mating, during mating, and up to Day 17 of gestation.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

10 mg/kg bw/day

Doses / concentrations 3

Dose / conc.:

100 mg/kg bw/day

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day

No. of animals per sex per dose:

44 (22 males and 22 females)

Control animals:

yes, concurrent vehicle

Positive control:

none

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: twice weekly (males), on Gestation Days 0, 3, 7, 10, 14, 18, and 20 (females)

FOOD CONSUMPTION AND COMPOUND INTAKE:
Prior to mating,food and water consumption for males and females was recorded weekly and daily, respectively. During the gestation
period, food and water consumption was measured only for females during the following time periods: Gestation Days 0 to 2, 3 to 6, 7 to 9,
10 to 13, 14 to 17, and 18 to 19, inclusive.

Oestrous cyclicity (parental animals):

10 days before the mating period began, vaginal smear samples were obtained daily from all females to assess the regularity, as well as the duration of estrous
cycles.

Sperm parameters (parental animals):

The left vas deferens was ligated to obtain a 5 μL sample from the cauda epididymis to assess for motility according to the following grades:
no sperm motile; few sperm motile; most sperm motile, slow moving; or most sperm motile, fast moving.

Postmortem examinations (parental animals):

Females:
- killed on Day 20 of gestation by carbon dioxide inhalation, and uterine contents examined
- each female macroscopically examined for evidence of disease or adverse reaction
- number of corpora lutea in each ovary counted
- reproductive tract, including ovaries dissected out
- for each female, the numbers of pre- and post-implantation sites, early and late resorption sites, and viable fetuses, as well as the distribution of fetuses in each uterine horn, examined
- uterus of any female presumed to be nonpregnant was stained using 10% aq (v/v) ammonium sulfide solution and examined for implantation sites

Males:
- reproductive organs weighed

Postmortem examinations (offspring):

- each fetus weighed, subjected to detailed external examination
- placental weights were recorded and examined macroscopically for any abnormalities
- neck, thoracic, and abdominal cavities were removed from half of the fetuses, the contents of the thoracic and abdominal cavities were examined, and sex was recorded
- fetuses subjected to a skeletal examination

Statistics:

To test the statistical significance of suggestive intergroup differences, one-way analysis of variance and t test were performed on body weights, body weight
changes, and food and water consumption. Organ weights were evaluated by Dunnett’s or Behren’s-Fisher’s tests. Nested analysis of variance and weighed
t test were conducted on fetal and placental weights. Differences with an associated probability of P<0.05 were deemed to be statistically significant.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Mortality:

no mortality observed

Description (incidence):

One male treated with 1000 mg/kg bw/day and demonstrating abdominal distension, pallor, ptosis, irregular respiration, and a decrease in body weight was killed during Week 6.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Details on results (P0)

- no differences observed in number of corpora lutea, pre- and postimplantation sites, early and late resorptions, and viable fetuses.
- absolute and relative weights of reproductive organs similar between treatment groups and control group
- evaluation of sperm number and motility revealed no findings attributable to treatment
- placental weights were not affected by treatment

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Remarks:

reproductive toxicity

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: highest dose tested

Target system / organ toxicity (P0)

Critical effects observed:

not specified

Results: F1 generation

Details on results (F1)

Upon macroscopic, internal, and skeletal examinations of the fetuses, no variations were observed that were not comparable to historical control values. There were no observed effects related to treatment. Fetal weights were not affected by treatment. Fetal sex ratios were comparable between all treatment groups and the control group.

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Remarks:

developmental toxicity

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: highest dose tested

Target system / organ toxicity (F1)

Critical effects observed:

not specified

Overall reproductive toxicity

Reproductive effects observed:

no

Any other information on results incl. tables

Table 1: Mean Reproductive parameters of rats treated with Behenyl alcohol

	Dose (mg behenyl alcohol/kg body weight)
	0	10	100	1000
Number of pregnant animals	22	22	22	21
Corpora lutea count	17.8 (2.7)	18.4 (4.0)	18.7 (2.3)	18.9 (2.4)
Implantations	17.2 (2.6)	17.0 (3.2)	18.1 (1.8)	18.0 (2.3)
viable young male	8.4 (2.9)	8.4 (2.3)	8.5 (2.5)	8.6 (2.6)
viable young female	8.0 (3.0)	7.5 (2.6)	8.5 (2.2)	8.3 (2.2)
viable young total	16.4 (3.2)	15.9 (3.5)	17.0 (2.3)	16.9 (2.2)
early resorptions	0.82 (0.90)	1.09 (1.04)	1.14 (1.07)	1.05 (1.02)
late resorptions	0.00 (0.00)	0.0 (0.00)	0.00 (0.00)	0.00 (0.00)
total resorptions	0.82 (0.90)	1.09 (1.04)	1.14 (1.07)	1.05 (1.02)
Pre-Implantation loss (%)	3.3	8.3	3.2	5.8
Post-Implantations loss (%)	4.7	6.4	6.3	5.8

Numbers in parentheses represent standard deviations.