tert.butyl amine salt of (+)-(3R, 5S)-7[4-(4-Fluorophenyl)-6-Isopropyl-2 [methyl-(methylsulfonyl amino] pyrimidin-5-yl]-3,5- dihydroxy-6(E)- heptenoic acid

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

tert.butyl amine salt of (+)-(3R, 5S)-7[4-(4-Fluorophenyl)-6-Isopropyl-2 [methyl-(methylsulfonyl amino] pyrimidin-5-yl]-3,5- dihydroxy-6(E)- heptenoic acid

Cas Number:

917805-74-4

Molecular formula:

C26H39FN4O6S

IUPAC Name:

tert.butyl amine salt of (+)-(3R, 5S)-7[4-(4-Fluorophenyl)-6-Isopropyl-2 [methyl-(methylsulfonyl amino] pyrimidin-5-yl]-3,5- dihydroxy-6(E)- heptenoic acid

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS

- Source: TOXI COOP ZRT. (Budapest, Cserkesz street 90.)
- Age at study initiation: 8 weeks
- Fasting period before study: overnight
- Housing: 3 animals/ cage Type II polypropylene/polycarbonate
- Bedding: Laboratory bedding
- Diet (e.g. ad libitum): ad libitum, ssniff R/M-Z+H
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days in first step, 8 days in second step, 14 days in third step and 15 days in fourth step


ENVIRONMENTAL CONDITIONS

- Temperature: 22+/-3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 8-12/hour
- Photoperiod (hrs dark/ hrs light): 12 h dark/ 12 h light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: oleum helianthi

Details on oral exposure:

VEHICLE

- Conentration in vehicle: 200 and 30 mg/ mL
- Amount of vehicle (if gavarage): 10 mL/kg bw

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/ kg bodyweight

Doses:

2000 mg/kg
300 mg/kg

No. of animals per sex per dose:

2*3 females (2000 mg/kg)
2*3 females (300 mg/kg)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations: individually after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and twice each day for 14 days thereafter
- Frequency of weighing: just before treatment, on the 7th and 15th day
- Necropsy of survivors performed: yes
- Other examinations performed: individual observations included the status of skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic, central nervous system and somatomotor activity as well

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Mortality:

All rats dosed at 2000 mg/kg Rosuvastatin TBA Salt (EGIS-14660-3) died between the Day 1 and Day 11. One-one animal died on Day 1 (No.: 3656), Day 4 (No.: 3655), Day 6 (No.: 3641) and Day 11 (No.: 3653) and two animals died on Day 10 (No.: 3643, 3647). All deaths seemed to be consequences of systemic toxic effect of the test item.

No death occurred at 300 mg/kg single oral dose of the test item.

Clinical signs:

In group 1 treated with 2000 mg/kg dose clinical sign of reaction comprised of decreased activity (3 cases out of 39 observations), abnormal gait (5/39), incoordination (1/39), decreased righting reflex (4/39), decreased grip- and limb tone (3/39), decreased body tone (12/39), closed eyes (1/39), piloerection (9/39), decreased plantary reflex (2/39), decreased abdominal tone (2/39). Decreased activity (score -1, -2, -3), abnormal gait ( score +1, +2), decreased righing reflex (score -1, -2), decreased grip- and limb tone (score -1, -2), decreased body tone (score -1, -2) and piloerection (score +1, +2, +3) occurred in all animals. Decreased plantary reflex (score -1) and decreased abdominal tone (score -1) were found in two animals (No.: 3643, 3647). However, incoordination (score +3) and closed eyes (score +2) were observed in animal No.: 3641.
These symptoms occurred between Day 5 and Day 9. All animals were free symptoms on the treatment day and between Day 1 and Day 4.



In group 2 treated with 2000 mg/kg dose clinical sign of reation comprised of decreased activity (9 cases out of 28 observations), abnormal gait (4/28), incoordination (2/28), decreased body tone (9/28), blood around the eyes (4/28) and piloerection (6/28), Decreased activity (score -1, -2), decreased body tone (score -1, -2) and blood around the eyes (score +2), occurred in two animals (No.: 3653, 3655). However, abnormal gait (score +1, +2), incoordination (score +1) and piloerection (score +1) were observed in animal No.: 3653.
These symptoms occurred between Day 2 and Day 10. All animals were free of symptoms on the treatment day. Besides, animal No.:3655 had no any symptoms on Day 0 and Day 1 and animal No.: 3653 was free of symptoms between Day 0 and Day 2, respectively.

In group 3 and 4 treated with 300 mg/kg dose, the animals were free of symptoms during the study.

Body weight:

In group 1 (2000 mg/kg) two animals lost body weight between treatment day and Day 7. The loss in body weight was 50 g (24%) in animal No.: 3643 and 43 g (20%) in animal No.: 3647)

In group 2 (2000 mg6kg) one animal lost body weight between treatment day and Day 7. The loss in body weight was 44 g (21 %) in animal No.: 3653.

The body weight loss can be related to the test item toxic effect. The body weight and body weight gain data of the other animals inside the 2000 mg/kg dose group could not be evaluated in consequence of mortalities.

In group 3 and 4 (300 mg/kg), the mean body weight of the animals corresponded to their species and age throughout the study.

Gross pathology:

All rats treated with 2000 mg /kg dose of the test item spontaneously died during the study. There was no any macroscopic alteration in all died animals treated with 2000 mg/kg dose. All animals treated with 300 mg/kg dose of test item survived until the scheduled necropsy on Day 15. Moderate hydrometra observed in female No.: 3648 of the group 3 (300 mg/kg) is physiological finding and connected to the cycle of the animal. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The method used, is not intended for the precise calculation of a precise LD50 value.
The test item is ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as GHS category 4.