Formamide, N-[(2-bromo-5-hydroxy-4-methoxyphenyl)methyl]-N-[2-(4-hydroxyphenyl)ethyl]-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-10-24 to 2005-11-15

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

yes

Remarks:

limited information on test item, test animals and test conditions

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: SPL Standard Test Method 595.13.

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 has not been audited by the QA unit. No formal claim of GLP compliance is made for this study.

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

N-[(2-bromo-5-hydroxy-4-methoxyphenyl)methyl]-N-[2-(4-hydroxyphenyl)ethyl]formamide

Cas Number:

122584-18-3

Molecular formula:

C17H18BrNO4

IUPAC Name:

N-[(2-bromo-5-hydroxy-4-methoxyphenyl)methyl]-N-[2-(4-hydroxyphenyl)ethyl]formamide

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): JNJ-28645409-AAA (T002078)
- Physical state: solid (powder)
- Appearance: White, brown grey powder

Specific details on test material used for the study:

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
- Weight at study initiation: 18-23 grams

ENVIRONMENTAL CONDITIONS: no data

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

as a solution in dimethyl sulphoxide at concentrations of 50%, 25% and 10% w/w
A further group of four animals was treated with dimethyl sulphoxide alone.

No. of animals per dose:

four/dose group

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: no data
- Irritation: no data
- Lymph node proliferation response: no data
- Systemic toxicity: Following a preliminary sighting test, no signs of systemic toxicity were observed at a concentration of 50% w/w in dimethyl sulphoxide.
- Ear thickness measurements: no data
- Erythema scores: no data

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay
- Criteria used to consider a positive response: Test-to-control ratio greater than 3.0 indicates a positive result.

TREATMENT PREPARATION AND ADMINISTRATION:
- 50 uL (25 uL per ear) of the test substance as a solution in dimethyl sulphoxide at concentrations of 50%, 25% and 10% w/w was applied to the dorsal surface of the ear.
- A further group of four animals was treated with dimethyl sulphoxide alone.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

other: 2,4 Dinitrobenzene sulphonic acid, sodium salt (DNBS): 1%, 10%, 20% v/v in 1% pluronic L92 in distilled water

Statistics:

no data

Results and discussion

Positive control results:

see 'Any other information on results incl. tables'

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

1.31

Test group / Remarks:

10% (w/w) in dimethyl sulphoxide

Results 2

Parameter:

SI

Value:

1.1

Test group / Remarks:

25% (w/w) in dimethyl sulphoxide

Results 3

Parameter:

SI

Value:

1.15

Test group / Remarks:

50% (w/w) in dimethyl sulphoxide

Cellular proliferation data / Observations:

CELLULAR PROLIFERATION DATA
DPM/node obtained by dividing the DPM value by 8 (total number of lymph nodes)
vehicle: 1248.42
10% (w/w) group: 1440.78
25% (w/w) group: 1377.83
50% (w/w) group: 1632.01

DETAILS ON STIMULATION INDEX CALCULATION
The Stimulation Index (SI) was expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group

EC3 CALCULATION
Not applicable (SI < 3.0)

CLINICAL OBSERVATIONS:
no signs of systemic toxicity

BODY WEIGHTS
The mean body weight gain shown by the animals over the study period was considered to be normal.

Any other information on results incl. tables

Positive Control Local Lymph Node Assay in the Mouse (2005)

Start Date	Finish Date	Test Material	Concentration	Vehicle	Stimulation Index^a	Classification^b
04/03/2005	10/03/2005	α‑Hexylcinnamaldehyde, tech., 85%	5%, 10%, 25% v/v	acetone/olive oil 4:1	2.76, 3.34, 8.91	Positive
20/04/2005	26/04/2005	α‑Hexylcinnamaldehyde, tech., 85%	5%, 10%, 25% v/v	ethanol/distilled water 7:3	2.64, 8.36, 12.94	Positive
14/07/2005	20/07/2005	α‑Hexylcinnamaldehyde, tech., 85%	5%, 10%, 25% v/v	1% pluronic L92 in distilled water	0.86, 1.50, 6.17	Positive
14/07/2005	20/07/2005	2,4 Dinitrobenzene sulphonic acid, sodium salt (DNBS)	1%, 10%, 20% v/v	1% pluronic L92 in distilled water	1.16, 9.59, 20.71	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 item was considered to be a non-sensitiser to the skin under the conditions of the test.