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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 November 2007 - 21 January 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: compliant to GLP and testing guideline; adequate coherence between data, comments and conclusions.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): LCA07012
- Substance type: mono-constituent
- Physical state: white powder
- Purity: not indicated
- Impurities (identity and concentrations): not indicated
- Purity test date: not indicated
- Lot/batch No.: 0715800012
- Expiration date of the lot/batch: 6 June 2010
- Stability under test conditions: not indicated
- Storage condition of test material: at room temperature, protected from sunlight and humidity

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeder
- Age at first treatment: approx. 10 weeks
- Weight at first treatment (mean): M=403 g, F=257 g
- Housing: individually, except during pairing
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Relative humidity: 50 ± 20%
- Light/dark cycle: 12h/12h
- Ventilation: 12 cycles/hour of filtered, non-recycled air.

IN-LIFE DATES: beginning: 27 November 2007 / end: up to 21 January 2008

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
test item was ground, mixed with vehicle heated to 45°C, forming a solution.
The test item dosage forms were prepared at up to 11-day intervals

VEHICLE
- Justification for use and choice of vehicle (if other than water): not indicated
- Concentration in vehicle: 5, 15 and 40 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg/day
- Lot/batch no. (if required): S 35142-236, S 35142-017 and S 39776-337
- Purity: not indicated
Details on mating procedure:
- M/F ratio per cage: 1
- Length of cohabitation: until mating occurred
- Proof of pregnancy: vaginal plug, or sperm in vaginal smear - referred to as day 0 post-coitum
- Mated for up to 14 days
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Description and results awaited from the study director
Duration of treatment / exposure:
from 2 weeks before mating until the end of mating (males: total of 37-38 days) or day 5 pp (females: total of 42-55 days)
Frequency of treatment:
once daily
Details on study schedule:
- no F1 parents (only one generation mated).
- Age at mating of the mated animals in the study: 13-16 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 25, 75, 200 mg/kg/day (m/f)
Basis:
other: nominal per gavage
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: in a previous 14-day toxicity study in the rat (see 7.5.1) mortality occured at 1000 mg/kg/day and moderate toxicity (limited to clinical signs and weight gain) occured at 300 mg/kg/day.
- Rationale for animal assignment: computerized stratification procedure (average body weight of each group is similar)
Positive control:
none

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule: day 1 and then weekly; and in F also days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum

FOOD CONSUMPTION for each animal: Yes
- Time schedule: weekly (last 7-day consumption period)

FOOD EFFICIENCY: No

WATER CONSUMPTION: No
Oestrous cyclicity (parental animals):
Yes : from a fresh vaginal lavage, each morning during the mating period, until the females were mated
Sperm parameters (parental animals):
No seminology examinations.
Testis and epididymis : weight (all males), microscopic exmaination (some rats: see table 1).
Litter observations:
STANDARDISATION OF LITTERS: No

PARAMETERS EXAMINED:
The following parameters were examined in offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight on days 1 and 5, clinical signs
Postmortem examinations (parental animals):
SACRIFICE
- All male survivors: after the end of the mating period
- All female survivors: on day 6 post-partum

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- Sacrifice on non-selected breeders/progeny: not applicable (only 1 generation of parents and of offspring)
- All pups sacrificed on day 5 post-partum

GROSS NECROPSY: Yes, on pups sacrificed and found dead
- external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGTHS: Not performed
Reproductive indices:
Pre-implantation loss = 100 * (Number of corpora lutea - Number of implantation sites) / Number of corpora lutea
Post-implantation loss = 100 * (Number of implantation sites - Number of live fetuses) / Number of implantation sites
Mating index = 100 * Number of mated animals / Number of paired animals
Fertility index = 100 * Number of pregnant female partners / Number of mated pairs
Gestation index = 100 * Number of females with live born pups / Number of pregnant females
Live birth index = 100 * Number of live born pups / Number of delivered pups
Offspring viability indices:
Viability index = 100 * Number of surviving pups on day 5 post-partum / Number of live born pups

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY
Loud breathing was observed at all dose-levels but no microscopic abnormalities were found in the respiratory organs of the 200 mg/kg/day animals therefore it was considered that this clinical observation did not induce long-term effects. There were no unscheduled deaths at any dose-level.

BODY WEIGHT GAIN:
A minimally lower mean body weight gain during the first 2 weeks of treatment of the males treated at 200 mg/kg/day, with recovery during the second 2 weeks of treatment.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Remarks:
parental toxicity
Effect level:
200 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: NOAEL = highest tested dose, without relevant effects.
Dose descriptor:
NOEL
Remarks:
reproduction (mating and fertility)
Effect level:
200 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: NOEL = highest tested dose, without relevant effects.

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Details on results (F1)

Pups from the groups treated at 25 or 75 mg/kg/day showed slightly lower mean body weight gains but there were no effects at 200 mg/kg/day.

Effect levels (F1)

Dose descriptor:
NOEL
Remarks:
offspring toxicity
Generation:
F1
Effect level:
200 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: NOEL = highest tested dose, without relevant effects.

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Not necessary

Applicant's summary and conclusion

Conclusions:
Under on the experimental conditions of this study, the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 200 mg/kg/day, the No Observed Effect Level (NOEL) for reproductive performance (mating and fertility) was considered to be 200 mg/kg/day and the NOEL for toxic effects on progeny was 200 mg/kg/day.
Executive summary:

The test item, LCA07012, was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before mating, during mating, gestation and until day 5 post-partum, at dose-levels of 25, 75 or 200 mg/kg/day.

Loud breathing was observed at all dose-levels but no microscopic abnormalities were found in the respiratory organs of the 200 mg/kg/day animals therefore it was considered that this clinical observation did not induce long-term effects. Males treated at 200 mg/kg/day gained minimally less weight during the first 2 weeks of treatment but some of this was recuped during the second 2 weeks and was therefore considered not to represent an effect of treatment. No histopathological lesions considered to be reflective of systemic toxicity were observed.