Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental start date: 17 october 2012 - Experimental completion date: 4 december 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD, GLP study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid - liquid: suspension
Details on test material:
Identification: Tritil-olmesartan medoxomil
Description: Powder
Lot/batch number: 2U003
Certificate of analysis: 11 september 2012
Storage conditions bulk material: room temperature
Purity: 99.7°C
Expiry date: april 2013

In vivo test system

Test animals

Species:
mouse
Strain:
other: Balb/c N
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories France, Les Oncins, BP0109, 69592 L'Arbresle Cedex, France
- Age at study initiation: approx. 8 to 12 weeks
- Weight at study initiation: approx. 20 +/- 4 g
- Total No. of animals on study: 4 animals/group
- Housing: Housed 4/cage in type "1290D" cages made of transparent polycarbonate. Space allocated: 820 cm² x 15.5 cm
- Diet (e.g. ad libitum): Irradiated controlled pelleted rodent diet (re. AO4C-10, SAFE)
- Water (e.g. ad libitum): free acess to tap water through bottles
- Acclimation period: for at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C to 24°C
- Humidity (%): 40 to 75 %
- Photoperiod (hrs dark / hrs light): 12h/12h cycle

Study design: in vivo (LLNA)

Vehicle:
dimethyl sulphoxide
Concentration:
25 %
No. of animals per dose:
4
Details on study design:
TREATMENT PREPARATION AND ADMINISTRATION:
- Day 1 to day 3: treatment (once daily): open application of 25 µL of the appropriate dilution of the test article or the positive control , or the vehicle alone to the dorsum of both ears.
- Day 4 and day 5: no treatment
- Day 6: injection (tail vein) of 250 µl of sterile phosphate-buffered saline (D-PBS) containing20 µCi of 3H-methylthymidine to all animals. Approximately five hours later, animals will be sacrified and the draining auricular lymph nodes excised and pooled in D-PBS for each experimental group.

Preparation of cell suspension
A single cell suspension of lymph node cells (LNC) is prepared for each pooled sample by gentle mechanical disaggregation through 100 µm Nylon Cell Strainer. The cells are washed twice with an excess of D-PBS and DNA precipitated with 5% trichloroacetic acid (TCA) for approximately 18 hours, in the refrigerator. After centrifugation, the supernatant is discarded and the pellet resuspended in 1mL TCA and transferred to vials, then 10 mL of scintillation fluid for 3H- counting are added.

Determination of incorporated 3H-methyl thymidine
Incorporation of 3H-methyl thymidine is measured by beta-scintillation counting as disintegrations per minute (DPM) for each experimental group (total DPM) with a beta-scintillation counter Wallac.

- Criteria used to consider a positive response: a test article is considered as a skin sensitizer when the stimulation index (SI) >= 3
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
No statistical analysis was performed.

Results and discussion

Positive control results:
HCA at 25% :
Total DPM/group: 17339.2;
Mean DPM/node: 2167.4;
Simulation index: 13.2
Acceptance criteria validated: SI is greater than 3.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: 1.2
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Total DPM/group: 8498.8 Mean DPM/node: 1062.4

Any other information on results incl. tables

No mortality was observed during the study.

No systemic toxicity or local irritation on application sites were noted in any of the treated animals.

Body weight variations in Tritil-olmesartan-medoxomil-treated group animals were similar to those observed in the vehicle control group animals.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In conclusion, under these experimental conditions, the test article Tritil-olmesartan-medoxomil was found negative in the local lymph node assay and was not considered as a skin sensitizer.