[1,1'-Biphenyl]-2-carbonitrile, 4'-[(1,4'-dimethyl-2'-propyl[2,6'-bi-1H-benzimidazol]-1'-yl)methyl]-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

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, EPA guidelines, see below).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

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: 17 October 2005
- Lot/batch No.: T02/03
- Expiration date of the lot/batch: April 2006
- Storage condition of test material: at room temperature , dark and dry

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

not specified

Details on test animals and environmental conditions:

TEST ANIMALS
-Source: LAB-ALL, Bt. Budapest, 1174 Hunyadi u. 7.
- Weight at study initiation: 313-349 g
- Housing: al animals were housed in Macrolon cages, size III, with 2-3 animals per cage.
- Diet (e.g. ad libitum): PURISTAR standard diet for rabbit ad libitum
- Water (e.g. ad libitum): tap water ad libitum, with 50 mg/100 mL ascorbic acid.
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3°C
- Humidity (%): 30 - 70% (relative humidity)
- Air changes (per hr): 8-12 per hour by central air-condition system
- Photoperiod (hrs dark / hrs light): 12 hours light daily from 6 am - 6pm (artificial light).

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: methyl cellulose (1%)

Induction 2

Concentration / amount:

BIBR 277 Nitril was formulated in methyl cellulose (1%). the mixture was stirred using a magnetic stirrer before and during the treatment


For intra-dermal exposure test: BIBR 277 Nitril was adminstrated at concentrations of 0.1%
For dermal induction: BIBR 277 Nitril was applied at concentrations of 75%
For the challenge exposure: BIBR 277 Nitril was applied at concentrations of 50% in vehicle (methyl cellulose (1% ))
Control animals were treated with vehicle methyl cellulose ( 1%)

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: methyl cellulose (1%)

Challenge 2

Concentration / amount:

BIBR 277 Nitril was formulated in methyl cellulose (1%). the mixture was stirred using a magnetic stirrer before and during the treatment


For intra-dermal exposure test: BIBR 277 Nitril was adminstrated at concentrations of 0.1%
For dermal induction: BIBR 277 Nitril was applied at concentrations of 75%
For the challenge exposure: BIBR 277 Nitril was applied at concentrations of 50% in vehicle (methyl cellulose (1% ))
Control animals were treated with vehicle methyl cellulose ( 1%)

No. of animals per dose:

10 test animals, 5 control animals per test.

Details on study design:

RANGE FINDING TESTS: a preliminary dose-finding study was performed
Test item was applied at concentrations of 0.01 and 0.1 % (w/v) for intra-dermal injection and 10, 25, 50 and 75% (w/v) for dermal application.

MAIN STUDY
A. INDUCTION EXPOSURE

- No. of exposures: 6 injections per animal for intra-dermal induction test, 1 per animal for dermal induction test.
- Exposure period: 24h (intradermal), 48h (epicutaneous)
-day(s) of challenge: day 8 after first induction test
- Test groups: 1 group, 10 animals per group
- Control group: 1 group, 5 animals per group
- Evaluation (hr after challenge): 24 or intra-dermal induction test and 1, 24, 48 and 72 hours after exposure



B. CHALLENGE EXPOSURE
- No. of exposures: 1 (epicutaneous)
- Day(s) of challenge: day 22 after first induction test
- Exposure period: 24 hours
- Test groups: 1 group, 10 animals per group
- Control group: 1 group, 5 animals per group
- Evaluation (hr after challenge): 24 and 48 hrs after exposure

Challenge controls:

Challenge controls were treated with the test substance at c=50%.

Positive control substance(s):

yes

Remarks:

Reliability studies are carried out twice a year, using items that are known to have moderate skin sensitisation properties. Latest check: 16 feb-12 March 2012 . Results are available.

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.1%

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.1%. No with. + reactions: 0.0. Total no. in groups: 10.0.

Results 2

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

none

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: none. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 3

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

75%

No. with + reactions:

0

Total no. in group:

19

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 75% . No with. + reactions: 0.0. Total no. in groups: 19.0.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

vehicle only

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle only. No with. + reactions: 0.0. Total no. in groups: 5.0.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Challenge with the test item BIBR 277 Nitril caused no signs of sensitisation in test animals previously sensitised. At the same time, none of the control animals showed signs of skin sensitisation. The net response value was thus 0%, as no indications for adverse skin reactions were noted.
The test item was classified as a non-senisitiser, based on this guinea pig test and according to current EU-regulations.