Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Start of experimental phase: 09 October 2008 - End of experimental phase: 14 November 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study according to international guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to
Guideline:
other: The Rules governing Medicinal Products in the European Community, Vol. 3B (1998)
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Name: KETOPROFEN/CFPPN
Synonym: CFPPN-Methylester
Batch number: F-PPR-080059
Expiry date: 12 August 2009
Description: Brown liquid
Storage at RTC : +4°C, protected from light (shipped at room temperature)
RTC reference number: 11437
Certificate of analysis : A certificate of analysis was supplied by the Sponsor (see Attachment I).
Test item characterisation: Not undertaken at the testing facility. The determination of the identity, strength, purity, composition, stability and method of synthesis and/or derivation of the test item was the responsibility of the Sponsor.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
Animal supply and acclimatisation
Species and strain: Rat, Hsd: Sprague Dawley SD
Number and sex: Females (nulliparous and non-pregnant)
Age and weight range (at order): 6 to 7 weeks old, 150 to 174 grams
Supplier: Harlan Italy S.r.l., 33049 San Pietro al Natisone (UD), Italy
Breeder: Harlan Italy S.r.l., 33049 San Pietro al Natisone (UD), Italy
Date of arrival: 25 September 2008
Weight range at arrival: 150.3 to 180.0 grams
Acclimatisation period: At least 5 days
Veterinary health check: after arrival

Caging
No. of animals/cage: 3 during the study; up to 5 during acclimatisation
Housing: Polycarbonate cages measuring 42.5x26.6x18 cm (step 1 to 4) or 59x38.5x20 cm (during acclimatisation), with stainless steel mesh lid and floor.
Cage tray control: Daily inspected and changed as necessary (at least 3 times/week)

Water and diet
Water: drinking water supplied to each cage via a water bottle
Water supply: ad libitum
Diet: 4 RF 18 (Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI) Italy)
Diet supply: ad libitum throughout the study except for dosing procedure


Housing conditions (parameter set)
Room lighting: Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours
Air changes: Approximately 15 to 25 air changes per hour
Temperature range: 22°C ± 2°C
Relative humidity range: 55% ± 15%
Actual conditions were monitored and recorded, and records retained. No relevant deviations occurred.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Remarks:
Concentration : 30 and 200 mg/ml
Details on oral exposure:
Dosing
Frequency of treatment : Once only, on the day of dosing (Day 1).
Fasting procedure : Overnight prior to dosing (Day –1) up to 4 hours after dosing.
Dose calculation : Dose volume of 10 ml/kg of body weight for each animal.
Dosing method : By gavage, using a rubber catheter attached to a graded syringe.
Doses:
Since information available suggested possible mortality occurring at the dose level of 2000 mg/kg, a first sub-group of 3 female animals was dosed at a level of 300 mg/kg (step 1). Mortality did not occur. A second subgroup, similarly composed, was then dosed at the same dose level (step 2). No mortality occurred. Consequently, a first subgroup of 3 females was dosed at 2000 mg/kg (step 3). Mortality did not occur. A second sub-group, similarly composed, was then dosed at the same dose level (step 4). No mortality occurred and no further doses were investigated since the objective of the study had been achieved.
No. of animals per sex per dose:
3 female animals/subgroup
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: mortality and morbidity: twice daily; clinical signs: day of dosing (on dosing, approximately 0.5, 2 and 4h after dosing), daily thereafter (14 days); body weight: allocation (day-1), days 1,2,8 and 15.
- Necropsy of survivors performed: yes

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed in the female animals dosed at 300 mg/kg (steps 1 and 2).
No mortality was recorded in the female animals dosed at 2000 mg/kg (steps 3 and 4).
Clinical signs:
At 300 mg/kg, reduced activity and piloerection were observed on the day of dosing. Recovery occurred by Day 2.
In the first subgroup of 3 animals (step 3), piloerection, reduced activity and hunched posture were observed on the day of dosing. Piloerection was then observed from Day 2 to Day 6 in the 3 animals and in a single animal on Days 7 and 8. Hairloss on the sacral region was observed in a single animal from Day 6 to the end of the obervation period.
In the second subgroup of 3 animals (step 4) piloerection was observed from Day 1 to Day 3. Hairloss on the sacral region was observed in two animals from Day 3 to Day 13. Recovery was completed by Day 14.
Body weight:
In animals dosed at 300 mg/kg changes in body weight were within the expected range for this strain and age of animals.

In animals dosed at 2000 mg/kg, a slight body weight loss was recorded on Day 15 in 3 animals dosed at 2000 mg/kg (1 animal of step 3 and 2 animals of step 4). This loss is likely due to the achievement of the plateau growth rather than to a toxicological effect. Body weight changes at the end of the observation period were within the expected range for this strain and age of animals.
Gross pathology:
No abnormalities were observed at necropsy examination performed at the end of the observation period on all the animals dosed at 300 mg/kg (steps 1 and 2) or 2000 mg/kg (steps 3 and 4).

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
These results indicate that the test item, KETOPROFEN/CFPPN, has a slight toxic effect on the rat following oral administration of a single dose at a level of 2000 mg/kg. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg body weight.

Mortality did not occur and some clinical signs, essentially reversible, were observed in the animals following dosing at 300 mg/kg or 2000 mg/kg, with a faster recovery period in the former case.

These results indicate that the test item, KETOPROFEN/CFPPN, has a slight toxic effect on the rat following oral administration of a single dose at a level of 2000 mg/kg. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg body weight.