Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 July - 22 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted in GLP compliance and in accordance with several internationally established guidelines (OECD, EEC guidelines, see below).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
not specified
GLP compliance:
yes (incl. QA statement)
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
480-420-2
EC Name:
-
Cas Number:
144702-27-2
Molecular formula:
C33 H29 N5
IUPAC Name:
4'-{[4-methyl-6-(1-methyl-1H-1,3-benzodiazol-2-yl)-2-propyl-1H-1,3-benzodiazol-1-yl]methyl}-[1,1'-biphenyl]-2-carbonitrile
Test material form:
other: solid
Details on test material:
- Name of test material (as cited in study report): BIBR 277 Nitril
- Physical state: Solid
- Analytical purity: 99.7%
- Purity test date: 20 March 2007
- Lot/batch No.: 80A
- Expiration date of the lot/batch: 08 March 2008
- Stability under test conditions: yes
- Storage condition of test material: At room temperature, dark and dry

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Budapest, Hungary
- Age at study initiation: 8 weeks
- Weight at study initiation: 180-272 g
- Housing: groupa caging (5 animals per cage)
- Diet (e.g. ad libitum): ssniff (R) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 36 - 69%
- Air changes (per hr): 8-12
- Photoperiod (hrs dark / hrs light): 12 hrs light daily from 6am - 6pm

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was prepared in PEG400 (50% of the final volume) and diluted with distilled water to the final volume at constant dosing volume of 10 mL/kg b.w.

VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg b.w.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analytical control of dosing preparations was performed during the 1st and 4th dosing weeks. The measured concentrationa ranged from 93% - 100% of the nominal concentrations.
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
25 mg/kg b.w.
Basis:
nominal in water
Remarks:
Doses / Concentrations:
100 mg/kg b.w.
Basis:
nominal in water
Remarks:
Doses / Concentrations:
300 mg/kg b.w.
Basis:
nominal in water
No. of animals per sex per dose:
each 5 per sex and dose,
5 per sex and control
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: doses were determined during a preliminary study
Positive control:
none

Examinations

Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

FOOD EFFICIENCY:
food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: 1 day after the last treatment
- Anaesthetic used for blood collection: Yes (euthanyl)
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: 1 day after the last treatment
- Anaesthetic used for blood collection: Yes (euthanyl)
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table were examined.

URINALYSIS: NO

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at the end of the 4th exposure week
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity

Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table)
HISTOPATHOLOGY: Yes (see table)
Other examinations:
weight of selected organs was checked
Statistics:
Statistic were performed using SPSS PC+ software for the following data:
bodyweight
clinical chemistry
haematology
organ weight data

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
no mortality occured in any of the animals. The general physical state of all testes animals was considered regular throughout the test. High dose animals (f/m) displayed thin faeces.
Mortality:
no mortality observed
Description (incidence):
no mortality occured in any of the animals. The general physical state of all testes animals was considered regular throughout the test. High dose animals (f/m) displayed thin faeces.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
minor variations were observed, however, these were not related to treatment due to the lack of dosage-relationship and the inconsistency between sexes.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
no siginificant changes in food intake compared to that of the control animals were noted.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Aenemia was observed in both f/m at the 300 mg/kg b.w. dosage level. In high dose females, RBC count and HGB content was slightly lower than in historical data. In correlation, significantly higher RDW value was observed in the high dose females.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
No siginificant observations were made in males. In females, there was a dose-dependant increase in cholesterol. Differences in total biliruin at the 300 mg/kg b.w. /d level correlated with haematological findings indicated a haemolytic origin of aenemia.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
Slight variations were noted, however, these were consideres as regular biological variations in the animal behaviour patterns.
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
liver weights were increased in high-dosed males and females. Spleen weight was increased in high-dosed females.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
no macroscopic findings were noted in any male animals. In females, pinprick-sized haemmorraghes
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no consistent microscopic changes observed.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
There were no consistent microscopic changes observed.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects
Dose descriptor:
NOEL
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
It is concluded that administration of BIBR 277 Nitril to Wistar Rats at dpsages of 25, 100 and 300 mg/kg bodyweight for 28 days was generelly well tolerated. The marked clinical pathology and organ weight changes seen in females at the highest dose indicated slight aenemia and functional hepatic changes without any associated histopathological changes. Trends observed in 300 mg/kg b.w. dosed male rats were comparable. Similarly, satistical slight differences in females given 100 mg/kg b.w. were all within the respective historic control ranges and were not considered to represent adverse effects. The NOAEL was determined to be 100 mg/kg bw/day and the NOEL was determined to be 25 mg/kg bw/day.