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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004-11-01 to 2004-12-07
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted following SPL Standard Test method 595.12.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: SPL Standard Test Method 595.12
Principles of method if other than guideline:
The assay determines the level of lymphocyte proliferation in lymph nodes draining the application site of the test material. Determination of lymphocyte proliferation is quantified by measuring the incorporation of radiolabeled thymidine in the dividing lymph node cells. The proliferation response of lymph node cells was expressed as the number of radioactive disintegrations per minute per lymph node (dpm/node) and as the ratio of 3HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes (Stimulation Index).
GLP compliance:
no
Remarks:
The study was conducted in a facility operating to Good Laboratory Practice within the UK national GLP monitoring programme, but the study report had not been audited by the QA Unit. No formal claim of GLP compliance was made for the study.
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
4-[(4-CHLOROPYRIMIDIN-2-YL)AMINO]BENZONITRILE
EC Number:
607-384-4
Cas Number:
244768-32-9
Molecular formula:
C11H7ClN4
IUPAC Name:
4-[(4-CHLOROPYRIMIDIN-2-YL)AMINO]BENZONITRILE
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study reports): JNJ-4754724-AAA (T002488)
- Physical state: solid
- Appearance: light brown powder
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Janssen Pharmaceutica N.V.
- Expiration date of the lot/batch: No data
- Purity test date: No data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: No data
- Stability under test conditions: No data


TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No data.

In vivo test system

Test animals

Species:
mouse
Strain:
CBA/Ca
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: 15 to 22 grams
- Housing: No data
- Diet (e.g. ad libitum): No data
- Water (e.g. ad libitum): No data
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS:
- Temperature (deg C): No data
- Humidity (%): No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data

Study design: in vivo (LLNA)

Vehicle:
dimethylformamide
Concentration:
vehicle alone, 2.5%, 5% and 10% w/w
No. of animals per dose:
4
Details on study design:
RANGE FINDING TESTS:
- Following a preliminary sighting test, no signs of systemic toxicity were observed at a concentration of 10% w/w.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: a stimulation index of greater than 3.0

TREATMENT PREPARATION AND ADMINISTRATION:
- Three groups, each of four animals were treated with 50 µL (25µuL per ear) of the test substance as a solution in dimethyl formamide at concentrations of 2.5%, 5% and 10% w/w. A further group of four animals was treated with dimethyl formamide alone.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
other: 2,4 Dinitrobenzenesulfonic acid, sodium salt at 1%, 10%, 20% v/v in 1% pluronic F-68 in distilled water
Statistics:
No statistical analysis was used.

Results and discussion

Positive control results:
see tables below

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Value:
1.81
Test group / Remarks:
4 animals in 2.5% w/w in DMF group
Parameter:
SI
Value:
2.25
Test group / Remarks:
4 animals in 5% w/w in DMF group
Parameter:
SI
Value:
1.91
Test group / Remarks:
4 animals in 10% w/w in DMF group
Cellular proliferation data / Observations:
CELLULAR PROLIFERATION DATA
DPM:
- 2.5% = 11089.82 (1386.23/node)
- 5% = 13787.05 (1723.38/node)
- 10% = 11719.21 (1464.90/node)
- Vehicle Only = 6129.19 (766.15/node)

EC3 CALCULATION
- Not applicable

CLINICAL OBSERVATIONS:
-No mortality or clinical observations were observed.

BODY WEIGHTS:
- No unusualy changes in body weight were observed.

Any other information on results incl. tables

Positive Control Local Lymph Node Assay in the Mouse (2004)

Start Date

Finish Date

Test Material

Concentration

Vehicle

Stimulation Indexa

Classificationb

29/04/2004

05/05/2004

α‑Hexylcinnamaldehyde, tech., 85%

5%, 10%, 25% v/v

acetone/olive oil 4:1

1.40, 2.23, 6.09

Positive

29/04/2004

05/05/2004

α‑Hexylcinnamaldehyde, tech., 85%

5%, 10%, 25% v/v

acetone/olive oil 4:1

1.74, 2.20, 8.89

Positive

14/10/2004

26/10/2004

α‑Hexylcinnamaldehyde, tech., 85%

5%, 10%, 25% v/v

tetrahydrofuran

1.97, 3.71, 7.82

Positive

29/09/2004

05/10/2004

2,4‑Dinitrobenzenesulfonic acid, sodium salt

1%, 10%, 20% v/v

1% pluronic F-68

in distilled water

1.03, 4.41, 13.55

Positive

27/10/2004

02/11/2004

α‑Hexylcinnamaldehyde, tech., 85%

10%, 25%, 50% v/v

cottonseed oil

1.52, 2.63, 5.07

Positive

 


a=        Ratio of test to control lymphocyte proliferation

b=        Stimulation index greater than 3.0 indicates a positive result

* =        Standard Test Method 595 (Pooled nodes)

·=        Standard Test Method 599 (Individual nodes)

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance was considered to be a non-sensitiser under the conditions of the test.