416-730-1

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

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

Adequacy of study:

key study

Study period:

2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: historical data are not given, food consumption data male animals third week are missing, missing details on mating procedure: how many attempts, replacement of male after unsuccesful mating and how females were caged after mating,

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SPF breeding, VELAZ s.r.o., Kolec u Kladna, Czech Republic, RCH CZ 21760152
- Age at study initiation: 10 weeks
- Weight at study initiation: males 307.89 - 309.22g, females: 224.33 - 226.55g
- Housing: exept from mating period, two rats of the same sex per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: males from: 19.01.2011 To: 22.02.2011 females: 19.01.2011 - 13.03.2011

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

suspension

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): daily just before administration
- Storage temperature of food: test item was administered by gavage

VEHICLE
- Amount of vehicle (if gavage): 10 ml/kg bw

Details on mating procedure:

- M/F ratio per cage: 1 to 1
- Length of cohabitation: 7d
- Proof of pregnancy: sperm in vaginal smear, referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

According to the information from Sponsor the main component of the test substance is insoluble in the used vehicle (water for injection).
Mixture of sample and deionised water results in yellow non-transparent suspension but undissolved particles of the test substance were easily visible in the application form and homogeneity could be checked by eye. Stability of the test substance in the application form cannot be verified analytically. The application form was prepared just before the application and there was no indication that test substance would have been unstable in the suspension in deionised water for that short time period.

Duration of treatment / exposure:

males and females - 2 weeks prior to the mating period and during the mating period,
pregnant females - during pregnancy and till the 3rd day oflactation,
males - after mating period - totally for 28 days,
nonpregnant females (mated females without parturition) - for 25 days after the confirmed mating

Frequency of treatment:

daily, 7d/week

Details on study schedule:

- Age at mating of the mated animals in the study: 12 weeks

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 160, 400 and 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on data of 14d range finder

Positive control:

no

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly
males - weekly
females - weekly in premating and mating period,
during pregnancy: day 0, 7th , 14th, 20th day, during lactation: 0. or 1st and 4th day;
pups (litters) - 0 or 1st and 4th day;

Sperm parameters (parental animals):

Parameters examined in P male parental generations:
testis weight, epididymis weight, prostate gland weight, pituitary glands weight, sperm motility, sperm morphology

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, presence of gross anomalies, physical or behavioural abnormalities

Dead pups were sexed and externally examined; the stomach was examined for the presence of milk. Pups killed on the 4th day of lactation were sexed and subjected to external examination of the cranium, and to macroscopic examination of the thoracic and abdominal tissues and organs.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals, males were killed at the end of the administration period - after 28 days of administration
- Maternal animals: All surviving animals, parental females were killed on the 4th day of lactation. Mated females without delivery were killed 26th day after confirmed mating

GROSS NECROPSY
- macroscopically examined for any pathological changes with special attention to the organs of the reproductive systems

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated below were prepared for microscopic examination and weighed, respectively:
relevant gross lesions, pituitary gland, coagulation gland, prostate gland, seminal vesicles, epididymis and testes (fixed in Davidson's suspension), cervix of uterus, ovaries, uterus and vagina

Postmortem examinations (offspring):

SACRIFICE
- sacrificed on day 4 of lactation

GROSS NECROPSY
- examination of thoracic and abdominal tissues and organs

Statistics:

The ANOVA test - Analysis of Variance (QC.Expert 2.5) at significance level 0.05 was used for the statistical analysis. This statistical analysis was used for the results of body weight, biometry of organs and number of pups. Control group with vehicle was compared with three treated groups. The results statistically significant on probability level 0.05 are indicated by figures with asterisk in the summary tables.

Reproductive indices:

Post-implantation loss, Pre-implantation loss, Post-natal loss, Male mating index, Female mating index, Male fertility index, Female fertility index

Offspring viability indices:

Gestation index, Survival index

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

yellow stained feces at mid and high dose groups

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

atrophic changes in prostate gland of high dose males

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)

- males: decrease of absolute and relative weight of prostate gland (doseindependent) was recorded in males of all dose levels. Changes did not exceed range of historical control. The histological examination demonstrated the increased occurrence of atrophic changes in prostate gland of males at the dose level I000 mg/kg/day. This finding was not dose dependend and is not considered to be an adverse effect.

- females: Dose dependent decreased relative weight of uterus was recorded in females at the dose level 1000 mg/kg/day (connection with oestrous cycle)

Effect levels (P0)

1

Dose descriptor:

NOEL

Effect level:

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

Sex:

male

Basis for effect level:

other: the value is based on the incidence of atrophic changes in prostate gland

2

Dose descriptor:

NOEL

Effect level:

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

Sex:

female

3

Dose descriptor:

NOAEL

Effect level:

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

Sex:

male/female

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Sex:

male/female

Overall reproductive toxicity

Reproductive effects observed:

not specified