Pregna-4,6-diene-3,20-dione, 17-(acetyloxy)-6-chloro-1-(chloromethyl)-, (1.alpha.)-

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September to October 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

1995-07-27

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

(1R,3aS,3bR,9S,9aS,9bS,11aS)-1-acetyl-5-chloro-9-(chloromethyl)-9a,11a-dimethyl-7-oxo-1H,2H,3H,3aH,3bH,7H,8H,9H,9aH,9bH,10H,11H,11aH-cyclopenta[a]phenanthren-1-yl acetate

EC Number:

605-617-4

Cas Number:

17183-98-1

Molecular formula:

C24 H30 Cl2 O4

IUPAC Name:

(1R,3aS,3bR,9S,9aS,9bS,11aS)-1-acetyl-5-chloro-9-(chloromethyl)-9a,11a-dimethyl-7-oxo-1H,2H,3H,3aH,3bH,7H,8H,9H,9aH,9bH,10H,11H,11aH-cyclopenta[a]phenanthren-1-yl acetate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Strain: HAN: WIST (SPF)
- Source: Schering AG
- Age at study initiation: not reported
- Weight at study initiation: males 220-276 g, females 195-225 g
- Housing: singly in conventional housing conditions (Makrolon type III cages)
- Diet and water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-24
- Humidity (%): 54-64
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.9 % NaCl + 0.085 Myrj 53

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The content of the test substance in the formulation was analytically verified for all doses at least at beginning and termination of the study.
Method of analysis: HPLC with UV absorption at 275 nm following appropriate dissolution and dilution with mobile phase appropriately.

Duration of treatment / exposure:

4 weeks

Frequency of treatment:

once daily

Doses / concentrations

Remarks:

Doses / Concentrations:
34, 174, 1000 mg/kg in an application volume of 10 ml/kg
Basis:
actual ingested

No. of animals per sex per dose:

6
except 34 mg/kg group: 7 males and 5 females

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes, all animals
All signs of weak health, reactions to treatment and behavioural changes were recorded.
- Time schedule: Animals were checked regularly twice daily, also on weekends.

BODY WEIGHT: Yes, all animals
- Time schedule for examinations: once weekly

FOOD CONSUMPTION:
- Time schedule for examinations: once weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

WATER CONSUMPTION: Yes, all animals
- Time schedule for examinations: once weekly

OPHTHALMOSCOPIC EXAMINATION: Yes, all animals
- Time schedule for examinations: week 4

HAEMATOLOGY: Yes, all animals, except for coagulation studies (only 2-6 males and 1-6 females)
- Time schedule for collection of blood: Day 24, blood for coagulation studies sampled on Day 24
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, no food was offered to the animals for at least 16 hours prior to blood sampling.
- Parameters checked: Hematological investigations including determination of the following parameters were performed on Day 24: erythrocyte and leucocyte count, hemoglobin, hematocrit (packed cell volume), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), reticulocyte count, platelets, and differential count [neutrophils (myelocytes, immature, band types l and II, segmented), Iymphocytes, eosinophils, basophils, monocytes].
Coagulation studies: The following parameters were determined in plasma from 2-6 male and 1-6 female animals per group on Day 24: thrombin time, thromboplastin time, activated partial thromboplastin time and fibrinogen.

CLINICAL CHEMISTRY: Yes, all animals
- Time schedule for collection of blood: Day 2, Day 24
- Animals fasted: Yes, no food was offered to the animals for at least 16 hours prior to blood sampling except for sampling on Day 2.
- Parameters checked: glutamic pyruvic transaminase (GPT) and alkaline phosphatase (AP) on Day 2 (approximately 24 hours after the first treatment) and Day 24, total cholesterol, glucose, urea nitrogen, total protein, protein electrophoresis, sodium, potassium, calcium and chloride on Day 24 only.

URINALYSIS: Yes, all animals
- Time schedule for collection of urine: Day 26
- Animals fasted: Yes, no food was offered to the animals for at least 16 hours prior to urine sampling
- Parameters checked: pH, specific gravity and volume as weil as protein, glucose, blood, ketones, urobilinogen, bilirubin and sediment of urine.

NEUROBEHAVIOURAL EXAMINATION: No

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Description (incidence):

One male rat (group 2, 15M) died because of application failure and one female (group 2, 23F) died because of arteriopuncture in blood sampling.

Food consumption and compound intake (if feeding study):

no effects observed

Ophthalmological findings:

no effects observed

Urinalysis findings:

no effects observed

Description (incidence and severity):

No compound-related effects were observed.
All findings observed in urine as weil as in urinary sediment were not considered to be compound-related because they were also seen in control animals or occurred sporadically.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed up to the highest dose

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

No animal died due to compound administration. One male rat (34 mg/kg group) died because of application failure and one female (34 mg/kg group) died because of arteriopuncture in blood sampling.

Up to and including the highest dose group no compound-related clinical signs, effects on water consumption, ophthalmoscopy, hematology, bone marrow, urinalysis, blood coagulation, organ weights or in gross and histopathology were observed.

From a dose of 174 mg/kg onwards, a slight but not significant increase in serum glucose was observed on day 24 in both sexes, but more pronounced in male rats. Additionally, after a dose of 1000 mg/kg, a slight but not significant decrease in food consumption and body weight gain was observed in the male rats. However, these effects were probably related to the slightly increased serum glucose level, which may have caused a decrease in appetite and which may indicate a reduced metabolic availability of glucose in the body. However, as all vaIues remained within the minimum-maximum range of reference data, all observed effects are regarded to be of no biological relevance.

Applicant's summary and conclusion

Conclusions:

The no-observable effect level of ZK 10882 in male and female rats after repeated (28-29 times) daily i.g. application was 34 mg ZK 10882/kg body weight.

The findings after repeated administration of daily dosages up to 1000 mg ZK 10882/kg did not indicate any clear-cut toxic effects of the compound.

Executive summary:

6 -Chlor-Chlormethyldien (ZK 10882) was administered daily via gavage to 7 -6 male and 6-5 female Wistar rats per dose group, in doses of 0, 34, and 1000 mg/kg body weight for a period of 4 weeks. The animals were regularly observed and weighed. Food and water intake was determined. Ophthalmology, hematology and clinical laboratory investigations on blood samples as well as urinalysis and investigations on bone marrow and blood coagulation were performed. Organs and tissues were subjected to gross and histopathological investigations and selected organs were weighed.

Survival was not affected by treatment and no adverse effects were detected up to and including the highest dose group.

Therefore, the NOAEL in male and female rats was established to be 1000 mg/kg body weight/day.