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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
Patch test on human volunteers
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
3 distincts periods on the protocol :
- Induction period : 9 consecutive applications, to the same area (0.02ml per volunteer, diluted to 5%, by occlusive epicutaneous route for about 48 hours or for the first two week-ends 72 hours, to the arm skin. Preparation being under a liquid form, was put onto a disc of filter paper inserted beforehand into cupule.
- Rest period : or incubation period during which the cells transformations possibly go on, leading to the modification or reactivity;
15 days without any application
- Challenge phase : singla application of about 0.02 ml per volunteer of the test article diluted to 5% by epicutaneous occlusive route for about 48 hours, to the back skin.

Cutaneous reaction, control of the sensitization was evaluated by the macroscopic examination of the reactions possibly noted about 24 and 48 hours after removal of the patches corresponding to the "challenge" application.
These examination were eprformed for the 1th, 8th (induction) and 10th (challenge) applications by comparison to the reactions possibly obtained with a patch alone (without test article), applied in parallel under the same conditions (negative control)

As regards evaluation of the sensitising potential, a reaction whose intensity is equal to 3 (erythema with infiltration, papulae, vesicles) was considered as "positive". If an individual irritation reaction had been noted during the first application, of after those corresponding to induction, or if an erythema was observed to the control area (right side), the test article was considered as "positive" if the challenge application had provoked a reaction whose intensity was clearly higher, and/ or if it had tented to increase as the readings were performed.
GLP compliance:
yes
Remarks:
Good Clinical practices
Type of study:
patch test
Justification for non-LLNA method:
As the study provided show no skin sensitising potential, and since this result is supported by public data and the assessment report of copper, no further testing are required, especially in vivo assays such as LLNA assay.

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
other: Human
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
65 volunteers of both sexes, healthy adults, were recruited and selected after a general medical examination taking into account the inclusion and non-inclusion criteria (which are presented hereafter), as well as the prohibition and restriction concepts defined in the study protcol.
All of the panellists were then finally included by the Investigator on the basis of a clinical examination specific to the study, performed just before its start, after signature of, among other things, the compensation modes form and of the informed consent statement : 2 of them abandoned during the induction phase (not linked to the test article applications).
Analysis of the results was thus made from a selection of 51 panellists composed of 46 (or 44) women and 7 men, from 21 to years old.

Inclusion criteria :
- Caucasian
- Age : from 18 to 70 years old
- Sex : Both
- Weight : included within the limits of the scale proposed by the Metropolitan Insurance Company
- Health condition : it correspond to the selection criteria defined in the procedures of I.E.C. Bulgarie, in order to eliminate, as much as possible, the volunteers incurring risks or presenting with redhibitory affections for the clinical studies performed by I.E.C. Bulgarie. These criteria were evaluated on the basis of questionnaires and clinical examinations listed in the protocol
- Volunteers with fixed abode
- Understanding of bulgarian language

Non-inclusion criteria :
- Volunteers not respecting rules related to the biomedical research
- Having participated in a sensitisation trial for 2 months and/ or in photo-irritation or photo-sensitisation study for 3 months
- Deprived from liberty by a judiciary or administrative decision, sick volunteers in situation of emergency
- Under age protected by law, as well as those admitted in sanitary or social facilities, ever since the research can be performed in another manner
- Having refused to give their agreement
- Having undergone organ excision, organ transplant, skull concussion with extended loss of consciousness for less than 5 years or with present after-effects
- Volunteers pregnant or breastfeeding mohthers, or not using medically acceptable contraceptive methods
- Having cardiovascular, pulmonar, digestive, neurologic, psychatric, genital, urinary, haematological, endocrine disorders, immunological deficit, an allergic background, a skin disease, a febrile illness
- Having intolerance to cosmetics, body hygiene or domestic products, to clothes, to products used at work such as colophane, rubber, nickel, aluminium
- Having had or being in the course of a long-term treatment
- Volunteers who recently sufferef from insolation, or having followed heliotherapy during the month preceding the start of the study
- Volunteers smoking more than 10 cigarettes a day and not accepting restriction of their tobacco consumption during the two days prior to the start of the study and throughout the whole study

Study design: in vivo (non-LLNA)

No. of animals per dose:
51
Details on study design:
Application of the test article were performed under occlusive patches, composed of an 8 mm-diameter aluminium cupule covering a contact surface of 50 mm2.
Preparation being under a liquide form, was put just before application, onto a disc of filter paper (7mm in diamter) inserted beforehand into the cupule.

About of 0.02 ml per volunteer of the test article diluted to 5% into distilled water (justification of the dose level : capacity of the cupule)

There is 3 phases :
- Induction period : application during 48 hours or 72 hours on the left area. A negative control is carried out on the right arm.
- Rest period : Volunteers were not submitted to any application from day 21 to day 35 for a 15 day period.
- "Challenge" phase : product is applied on the back during 48 hours. A negative control (patch alone, without test article) is also performed.
Challenge controls:
A negative control (patch alone, without test article) is also performed.
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.02 ml diluted to 5%
No. with + reactions:
0
Total no. in group:
53
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.02ml diluted to 5%
No. with + reactions:
0
Total no. in group:
53
Reading:
other: No positive control performed
Hours after challenge:
24
Group:
positive control
Dose level:
0
No. with + reactions:
0
Total no. in group:
0
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Remarks on result:
other: Number of volunteer in the negative group are not mentioned
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Remarks on result:
other: Number of volunteer in the negative group are not mentioned

Applicant's summary and conclusion

Interpretation of results:
other: Integrated approach
Conclusions:
Under the experimental conditions, single and repeated epicutaneous applications of this test article, diluted at 5%, under occlusive patch, in the healthy volunteer did not provoke any cutaneous sensitisation.
Human in vivo assays could not be used alone to set a classification for skin sensitisation but can be used on an integrated approach.
Therefore, based on an integrated approach, and under the tests conditions, the patch assay can support a non classificaqtion for skin sensitisation.