1,3-Dioxane-4-acetic acid, 6-[2-[2-(4-fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-[(phenylamino)carbonyl]-1H-pyrrol-1-yl]ethyl]-2,2-dimethyl-, 1,1-dimethylethyl ester, (4R,6R)-

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

tert-butyl 2-[(4R,6R)-6-{2-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-(propan-2-yl)-1H-pyrrol-1-yl]ethyl}-2,2-dimethyl-1,3-dioxan-4-yl]acetate

EC Number:

700-258-0

Cas Number:

125971-95-1

Molecular formula:

C40H47FN2O5

IUPAC Name:

tert-butyl 2-[(4R,6R)-6-{2-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-(propan-2-yl)-1H-pyrrol-1-yl]ethyl}-2,2-dimethyl-1,3-dioxan-4-yl]acetate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% methylcellulose in water for injections

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

analyses were performed by HPLC

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0 mg/kg/day
Basis:

Doses / concentrations 2

Remarks:

Doses / Concentrations:
250 mg/kg/day
Basis:

Doses / concentrations 3

Remarks:

Doses / Concentrations:
500 mg/kg/day
Basis:

Doses / concentrations 4

Remarks:

Doses / Concentrations:
1000 mg/kg/day
Basis:

No. of animals per sex per dose:

6 females + 6 males

Control animals:

yes, concurrent no treatment

Details on study design:

satellite control 6 females + 6 males
S 0mg/kg/day
S 1000 mg/kg/day

Examinations

Observations and examinations performed and frequency:

Experimental data collection
Body weight weekly
Food consumption weekly
Water consumption twice a week
Health condition daily
General clinical observation daily
Clinical observation battery weekly
Functional observations in the last week of administration
Haematological examination 29th (main groups) and 43 rd day of study (satellite groups)
Clinical biochemistry 29th and 43rd day of study
Urinalysis 28th and 42nd day of study
Biometry of organs 29th and 43rd day of study
Pathological examination 29th and 43rd day of study

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Water consumption and compound intake (if drinking water study):

effects observed, treatment-related

Haematological findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Details on results:

Results
After completion of all examinations the following effects of test substance were
distinguished:
No treatment-related effects were detected in the following parameters:
- health condition
- behavioral assessment
- functional observation
- mortality
- fundamental histopathological findings of observed organs.

Treatment-related findings observed were as follows:
Dose level 1000 mg/kg/day
Reversible changes:
- increased value of lymphocytes in females - statistically significant
- increased count of platelets in both sexes — not statistically significant
- increased value of protrombin time in males — not statistically significant
- increased count of trachea inflammation in both sexes
- increased count of desquamation of urinary bladder

Irreversible changes:
- increased value of protrombin time in females — statistically significant
- decreased absolute weight of liver in females - statistically significant
- decrease of body weight in both sexes — not statistically significant
- decrease of food consumption in both sexes - not statistically significant
- decrease of food conversion in both sexes - not statistically significant
- decrease of water consumption in both sexes
- decreased value of tromboplastin time in males — not statistically significant
- increased value of tromboplastin time in females — not statistically significant
- decreased values of glucose in blood in females — not statistically significant decreased
-decreased absolute weight of testes and epidydimides in males — not statistically significant
- increase of relative weight of brain and heart in females — not statistically significant

Protracted changes (observed oniy after 14 day survival- recovery period):
- decreased value of monocytes in both sexes - statistically significant
- increased value of monocytes in satellite animals in both sexes — statistically significant
- decreased value of lymphocytes in satellite males - statistically significant
- increased value of granulocytes in satellite males - statistically significant
- decreased values of calcium and phosphorus in blood in satellite males — statistically significant
- increased relative weight of kidney in males—not statistically significant
- decreased relative weight of kidney in satellite males — not statistically significant

Dose level 500 mg/kg/day
Changes:
- increased value of granulocytes and monocytes in males - statistically significant
- decreased value of lymphocytes in males - statistically significant
- decreased value of lymphocytes in females — not statistically significant
- decreased value of granulocytes in females — not statistically significant
- decreased value of tromboplastin time in males — not statistically significant
- increased value of tromboplastin time in females — not statistically significant
- decreased value of protrombin time in both sexes — not statistically significant
- increased value of ALP in blood of males — not statistically significant
- decreased absolute weight of heart in males — not statistically significant
- decreased absolute weight of spleen in females — not statistically significant
- increased absolute weight of uterus in females — not statistically significant
- increased absolute weight of adrenal glands in females — not statistically significant
- increased relative weight of testes in males—not statistically significant
- increased relative weight of uterus in females — not statistically significant
- decreased relative weight of kidney in females — not statistically significant
- increased count of trachea inflammation in both sexes
- increased count of stomach inflammation in males.

Dose level 250 mg/kg/day
Changes:
- decrease of body weight in both sexes — not statistically significant
- decrease of weight increment in both sexes - not statistically significant
- increased value of total count of erythrocytes in both sexes — not statistical, significant
- increased value of total count of leucocytes in females — not statistically significant
- decreased value of tromboplastin time in males—not statistically significant
- increased value of tromboplastin time in females — not statistically significant
- increased value of protrombin time in both sexes — not statistically significant
- decreased values of cholesterol in blood in females — not statistically significant
- decreased volume of urine and increased of urine pH in both sexes
- decreased absolute and relative weight of liver in both sexes — not statistically significant
- decreased absolute weight of kidney and thyrnus in males — not statistically significant
- increased absolute weight of ovary in females — not statistically significant
- decreased relative weight of thymus in males — not statistically significant
- increased relative weight of testes and brain in males — not statistically significant
- increased relative weight of ovary in females — not statistically significant.

Effect levels

Dose descriptor:

NOAEL

Effect level:

125

Sex:

male/female

Basis for effect level:

other: overall effects clinical signs; mortality; body weight; food consumption; water consumption and compound intake; haematology; clinical chemistry; urinalysis; gross pathology; organ weights; histopathology

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Conclusion
Overall assessment of the effects of test substance, Acetonide Atorvastatine, after 28 day gavage administration revealed that the substance caused no toxicologically significant changes in observed health condition parameters (health condition control, clinical observations, behavioral assessment, functional observation) and did not cause mortality or fundamental histopathological changes of observed organs in both sexes.
The test substance decreased body weight with keeping standard food consumption. It decreased absolute weight of liver, value of glucose in blood was lowered without morphological changes in liver.
Haematological examination showed primarily negative effect on leucocyte differential - increase of a number of granulocytes and monocytes with normal rate of total number of white blood cells. This change could indicate general response of organism to the toxic noxa. It was not accompanied by inflammatory changes in organs. These effects detected in the highest dose level 1000 mg/kg/day and in middle dose level 500 mg/kg/day were statistically significant and irreversible. At a dose level of 250 mg/kg/day no change in examined organs or parameters was statistically significant but for instance in haematology parameters some toxicologically important effects were detected. So this lowest dose level can not be established as the NOAEL (No Observed Adverse Effect Level, dose without toxic effects).
According to diminishing trend of changes in dose levels 1000-500-250 mg/kg/day it is possible to deduce, that the dose without unfavorable effects will be somewhere between about 100- 150 mg/kg/day and we estimate it to 125 mg/kg/day.
The dose level about 125 mg/kg/day could be regarded as the NOAEL (No Observed Adverse Effect Level).