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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2007-02-20 to 2007-03-16
Reliability:
1 (reliable without restriction)
Cross-reference
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
96/54/EEC
Deviations:
no
Principles of method if other than guideline:
NA
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test

Test material

Constituent 1
Details on test material:
NA

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
not specified
Details on test animals and environmental conditions:
Body weight range at starting: 285-326 g
Date of receipt: 31 January 2007
Acclimatisation time: 20 days

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: methyl cellulose (1%)
Concentration / amount:
On basis of results of the preliminary test item formulation evaluation and the Preliminary Dose Range Finding Study, the test item was used for dermal induction treatment at a concentration of 50 % (w/v). Control animals were treated with vehicle (methyl cellulose (1 %)) only. For intra-dermal treatment the test item was used at a concentration of 0.1 % (w/v). For the challenge exposure, all animals of the treatment and control group were treated with concentrations of 50 % (w/v) test item in vehicle.
Challengeopen allclose all
Route:
intradermal
Vehicle:
other: methyl cellulose (1%)
Concentration / amount:
On basis of results of the preliminary test item formulation evaluation and the Preliminary Dose Range Finding Study, the test item was used for dermal induction treatment at a concentration of 50 % (w/v). Control animals were treated with vehicle (methyl cellulose (1 %)) only. For intra-dermal treatment the test item was used at a concentration of 0.1 % (w/v). For the challenge exposure, all animals of the treatment and control group were treated with concentrations of 50 % (w/v) test item in vehicle.
No. of animals per dose:
Test group: 10 animals
Control group: 5 animals
Details on study design:
Main Study I: Intra-Dermal Induction Exposure
Before starting the exposure an area of approximately 5 x 5 cm on the scapular region of animals was clipped free of hair and shaven close with care.
Intra-dermal treatment
Test groups:
A series of three injections was administered on each side of treatment group animals, as follows, resulting in six injections per animal:
2 injections with 0.1 ml of Freund's complete adjuvant (FCA) mixed with physiological saline (1:1),
2 injections with 0.1 ml of the test item (0.1 %) homogenised in methyl cellulose (1 %),
2 injections with 0.1 ml of test item (0.1 %) mixed with physiological saline and homogenised in Freund's complete adjuvant (1:1).
Control group:
The control animals were treated similarly as the treatment group however, the vehicle, without the test item was used for injections, as follows:
2 injections with 0.1 ml mix of Freund's complete adjuvant and physiological saline (1:1)(v/v),
2 injections with 0.1 ml of methyl cellulose (1 %), 2 injections with 0.1 ml of 50 % w/v methyl cellulose (1 %) in a 1:1 mixture (v/v) of Freund's complete adjuvant and physiological saline.

Main study II: Dermal Induction Exposure
Six days after intra-dermal injections and 24 hours prior to topical induction application, all animals were exposed to 0.5 ml of 10 % sodium dodecyl sulphate in Vaseline, in order to create local irritation.
Approximately 24 hours after application, treatment animals were exposed to test item and control animals were treated with vehicle only.
Closed patch was fixed as follows: for treatment animals 0.5 ml of test item (at concentration of 50 % in 1 % methyl cellulose) was spread on the surface previously prepared and covered with a standard (5 x 5 cm) size porous gauze patch. The sample was secured by a piece of PVC tape and occluded with adhesive bandage.
Control animals were treated dermally with 0.5 ml of the vehicle methyl cellulose (1 %) and the dressing was prepared and applied as for treatment group animals.
The patches were left in place for 48 hours. At the end of exposure the bandage was removed and the excess material was wiped off.
Following dermal induction treatments the animals were left untreated for 14 days prior to challenge applications.
Challenge controls:
Main study III: Challenge Exposure
Two weeks after dermal treatment animals were exposed to dermal challenge applications. 24 hours before the challenge treatment left and right flank areas (5 x 5 cm) of each animal were prepared for application. Challenge application was carried out using the described closed patch technique. Left shaved flank areas of the animals (both of the treatment and control groups) were treated with 0.5 ml test item (at concentration of 50 % in 1 % methyl cellulose). The right shaved flank areas were treated with 0.5 ml methyl cellulose (1 %), in all cases.
Exposure time: 24 hours.
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole

Study design: in vivo (LLNA)

Concentration:
NA
No. of animals per dose:
NA
Details on study design:
NA
Statistics:
NA

Results and discussion

Positive control results:
On the basis of the present reliability study, reference item 2-Mercaptobenzothiazole caused positive responses in 60 % of the treatment group
animals.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: NA
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: NA

Any other information on results incl. tables

NA

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information criteria: EU
Conclusions:
The test item AVE 5530 EVA-KET was classified as non-sensitiser, based on this guinea pig maximisation test and according to current EU-regulations.
Executive summary:

Challenge with the test item AVE 5530 EVA-KET evoked no positive response in test animals previously sensitised. At the same time, none of the animals in the control group showed signs of skin sensitisation. The net response value or sensitisation rate was thus 0 %, as no indications for adverse skin reactions were noted.

The test item AVE 5530 EVA-KET was classified as non-sensitiser, based on this guinea pig maximisation test and according to current EU-regulations.