6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 May 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: done under GLP and OECD method

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Principles of method if other than guideline:

This study was performed to assess the skin sensitisation potential of CP-89,575 using the guinea-pig. The method foiiowed was that described in:EEC Methods for the determination of toxicity, Annex of Directive 92/69/EEC (OJ No. L383A, 29.12.92), Part B, Method B.6. Skin sensitisation.

MAGNUSSON, B. and KLIGMAN, A.M. (1970) Allergic Contact Dermatitis in the Guinea­ pig: Identification of contact allergens, Thomas, C.C., Springfield, Illinois, U.S.A.

Based on the results of a preliminary study and in compliance with the guideline, the following dose levels were selected:

Intradermal injection: 1% w/v in Alembicol D
Topical application: 50% w/v in Alembicol D
Challenge application: 50% and 25% w/v in Alembicol D

Ten test and five control guinea-pigs were used in this study.

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The LLNA method was not available in 1997 when this study was conducted.

Test material

Test material information

Constituent 1

Reference substance name:

6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

EC Number:

421-320-0

EC Name:

6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

Cas Number:

118289-55-7

Molecular formula:

Hill formula: C10 H9 Cl2 N O CAS formula: C10 H9 Cl2 N O

IUPAC Name:

6-chloro-5-(2-chloroethyl)-2,3-dihydro-1H-indol-2-one

Test material form:

other: solid

Details on test material:

Purity: 99.9%
Appearance: Off-white solid
Storage conditions: Room temperature in the dark
Date received: 2 February 1996

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source:D. Hall, Newchurch, Staffordshire, England.
- Age at study initiation: approximately four to five weeks of age.
- Weight at study initiation: weight range of 288 to 367 g on arrival
- Housing:The guinea-pigs were housed in groups of five in suspended metal cages with wire mesh floors in Building R 17 Room 14
- Diet (e.g. ad libitum): ad libitum, . Hay was given weekly.The batch of diet used for the study was not analysed for nutrients, possible contaminants or micro­ organisms.
- Water (e.g. ad libitum):A vitamin C enriched guinea-pig diet FD2 and drinking water were provided ad libitum
- Acclimation period: All the guinea-pigs were acclimatised to the experimental environment for five days prior to allocation to the main study.

- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):12/24

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other:

Induction 2

Concentration / amount:

Based on the results of the preliminary investigations, the following concentrations of CP-89,575 were selected:

Induction intradermal injection- 1% w/v in Alembicol D
Induction topical application- 50% w/v in Alembicol D
Topical challenge - 50% and 25% w /v in Alembicol D

lnjectables for the test animals were prepared as follows:

1. Freund's complete adjuvant** was diluted with an equal volume of water for irrigation (Ph.Eur.).

2. CP-89,575, 1% w/v in Alembicol D.

3. CP-89,575, 1% w/v in a 50 : 50 mixture of Freund's complete adjuvant and Alembicol D.

A 40 x 60 mm area of dorsal skin on the scapular region of the guinea-pig was clipped free of hair with electric clippers. Three pairs of intradermal injections were made into a 20 x 40 mm area within the clipped area

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other:

Challenge 2

Concentration / amount:

Based on the results of the preliminary investigations, the following concentrations of CP-89,575 were selected:

Induction intradermal injection- 1% w/v in Alembicol D
Induction topical application- 50% w/v in Alembicol D
Topical challenge - 50% and 25% w /v in Alembicol D

lnjectables for the test animals were prepared as follows:

1. Freund's complete adjuvant** was diluted with an equal volume of water for irrigation (Ph.Eur.).

2. CP-89,575, 1% w/v in Alembicol D.

3. CP-89,575, 1% w/v in a 50 : 50 mixture of Freund's complete adjuvant and Alembicol D.

A 40 x 60 mm area of dorsal skin on the scapular region of the guinea-pig was clipped free of hair with electric clippers. Three pairs of intradermal injections were made into a 20 x 40 mm area within the clipped area

No. of animals per dose:

Number of animals in test group: Ten (Animal Numbers) 950 to 959
Number of animals in negative control group: five control guinea-pigs were used in this study (Animal Numbers) 945 to 949

Details on study design:

MAIN STUDY

A. INDUCTION EXPOSURE
- No. of exposures: 2 - intadermal and topical
- Exposure period: 9 days total. Injection, 6 days later pretreatment with 10% sodium lauryl sulphare, 24 hrs later patch soaked in 0.4 ml of CP-89,575, 50% w/v in Alembicol D was placed on guinea pig for 48 hours.
- Control group: During the induction phase, the control animals were treated similarly to the test animals with the exception that the test substance was omitted from the intradermal injections and topical application


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: challenged topically two weeks after the topical induction application
- Exposure period: 24hr occlusive bandage
- Test groups:
- Control group:
- Site:left flank control; right flank test substance
- Concentrations:
- Evaluation (hr after challenge):24 and 48 hr after treatment- local reaction (erythema, edema)

OTHER:

Challenge controls:

no data

Positive control substance(s):

no

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

other:

Hours after challenge:

24

Group:

test chemical

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No signs of ill health or toxicity were recorded.

Remarks on result:

other: Reading: other:. . Hours after challenge: 24.0. Group: test group. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of ill health or toxicity were recorded. .

Results 2

Reading:

other:

Hours after challenge:

48

Group:

test chemical

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

No signs of ill health or toxicity were recorded.

Remarks on result:

other: Reading: other:. . Hours after challenge: 48.0. Group: test group. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of ill health or toxicity were recorded. .

Any other information on results incl. tables

Intradermal injections :

Necrosis was recorded at sites receiving Freund's Complete Adjuvant in test and control animals. Slight irritation was seen in test animals at sites receiving CP-89,575, I% w/v in Alembicol D and slight irritation was observed in control animals receiving Alembicol D.

Topical application :

Slight erythema was observed in test animals following topical application with CP-89,575, 50% w/v in Alembicol D. Slight erythema was seen in the control guinea-pigs

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

In this study, CP-89,575 did not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the ten test animals