Benzeneacetic acid, 4-[4-[4-(hydroxydiphenylmethyl)-1-piperidinyl]-1-butyn-1-yl]-.alpha.,.alpha.-dimethyl-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP, Guideline study. The study report pointed up two deviations from study plan but they were judged as minor deviations ((i) the name of the study director and (ii) the number of the test item because of a typing error) that do not affected the reliability of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

FEXO-07

IUPAC Name:

FEXO-07

Constituent 2

Reference substance name:

4-[ 4-[ 4-(1-hydroxydiphenylmethyl)-1-piperinidyl]- -1-butinyl]-a,a-dimethylbenzeneacetic acid

IUPAC Name:

4-[ 4-[ 4-(1-hydroxydiphenylmethyl)-1-piperinidyl]- -1-butinyl]-a,a-dimethylbenzeneacetic acid

Test material form:

other: white crystalline powder.

Details on test material:

Immediately upon receipt, the test item was registered, then stored at ambient temperature and protected from light.

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Albino Hartley

Sex:

male

Details on test animals and environmental conditions:

Body Weight: between 286.4 g an d 341.1 g at the start of the experiment.
Age: between 3 an d 6 weeks at the start of the experiment.
Number: (i) preliminary study: 6 males, (ii) main study: treated group of 10 males and a negative control group of 5 males and (iii) positive control group (at least every 6 months): 5 males. [The study involved 6 males far a preliminary test, and 15 males for the main study].
Allocation to treatment to each animal was randomly determined before the start of the study on the day prior to the first day of administration (D-1 ). Homogeneity of groups was validated on the criterion of body weight measured on the day of randomization. In each group, individual weights of the animals did not deviate from the group mean weight by more than ± 20%.
Housing: five males from each treatment group were kept in a cage of standard size, on dust-free white wood shavings (or equivalent) as bedding. The cages were cleaned according to the CERB SOP. The animals were placed in an air-conditioned (20 °C ± 3 °C) animal house kept at relative humidity between 45% and 65% (except during the cleaning slot) in which non-recycled filtered air which was changed approximately 10 times per hour. The artificial day/night cycle involved 12 hours light and 12 hours darkness (lights on at 7.30 a.m.).
Feeding: FD1 (P) SQC SDS/DIETEX feed distributed daily at fixed times. The criteria for acceptable levels of contaminants in the feed supplied were within the limits of the analytical specifications established by the diet manufacturer.
Drinking water: drinking water with 0.02% vitamin C added was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to the another laboratory for analysis. The criteria far acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.
Vehicle: sterile water was chosen an the basis of the physico-chemical characteristics of the test item.

Only healthy animals with an intact skin were used for the experiment.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: Sterile water

Induction 2

Concentration / amount:

Solid test item was tested in emulsion.
Concentrations were expressed as percentage weight of test item/weight of preparatian (w/w):
25% (w/w)
10% (w/w)
2.5% (w/w)
1% (w/w)
0.5% (w/w)
0.25% (w/w)
0.1% (w/w)
0.05% (w/w) were tested.

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: Sterile water

Challenge 2

Concentration / amount:

Solid test item was tested in emulsion.
Concentrations were expressed as percentage weight of test item/weight of preparatian (w/w):
25% (w/w)
10% (w/w)
2.5% (w/w)
1% (w/w)
0.5% (w/w)
0.25% (w/w)
0.1% (w/w)
0.05% (w/w) were tested.

No. of animals per dose:

2

Details on study design:

The test involved three subsequent treatments:
- intradermal induction: first contact between the body and FEX0-07
- topical induction: second contact with FEX0-07; in case of sensitising item recognition of allergen with multiplication of cellular and/or humoral transformation processes
- topical challenge: third contact with FEX0-07 (an area of skin having never previously been in contact with FEX0-07) leading to the occurrence of a cutaneous clinical manifestation if FEX0-07 has a sensitising potential.

Challenge controls:

A negative control group of 5 males has been chosen. (The control group was used to confirm that under the experimental conditions adopted, the appearance of skin reactions in the treated group in the absence of any reaction in the controls was due to an allergic response and not to skin irritation).

Positive control substance(s):

yes

Remarks:

Positive control group (at least every 6 months): 5 males.

Results and discussion

Positive control results:

In the positive control group in which animals were treated with HCA solution, 100% of animals showed skin reaction (score 1 to 2) at 24 and 48 hours.
(The positive test item HCA appeared as an extreme sensitiser).

Any other information on results incl. tables

Intradermal injections

- 2.5% (w/w) and l% (w/w) of FEX0 -07 in sterile water, any animal presented a discrete or patchy erythema (score l)

- 0.5% (w/w), 0.25% (w/w), 0.1% (w/w) and 0.05% (w/w) of FEX0-07 in sterile water, no skin reaction was observed in any animal - 2.5% (w/w) FEX0-07 diluted in sterile water, the injections were so difficult and the concentration was not also chosen for the main study.

Cutaneous application

First topical application: at 25% (w/w) and 10% (w/w) of FEX0-07 in sterile water, no skin reaction was noted in any animal.

Second topical application: at 25% (w/w) and 10% (w/w) of FEX0-07 in sterile water, no skin reaction was noted in any animal.

In conclusion:

- the maximum injectable concentration causing a moderate irritation tested by intradermal administration was 1% (w/w) of

FEX0-07 diluted in sterile water (it was used during the primary induction phase on D1)

- the maximum concentration causing a moderate irritation tested by cutaneous application was 25% (w/w) of FEX0-07 (it was used during the second induction phase or sensitisation phase on D9).

- the maximum non-irritant concentration (MNIC) determined by cutaneous application was 25% (w/w) of FEX0-07 diluted in sterile water (it was used in the challcnge phase on D22).

The following tables document the results that were judged more representative for this test report:

Table 2.5. Skin lesions - Individual scores - FEXO-07

                                                                                                                                                  

Observations (range of scores; normal score)	Time	Animal serial number
		1	2	3	4	5	6	7	8	9	10
Skin lesions(0 -3; 0)	day 24 (24 h)	0	0	0	0	0	0	0	0	0	0
	day 25 (48 h)	0	0	0	0	0	0	0	0	0	0

Table 2.7. Main study: Summary of results of cutaneous reactions: negative control and treated group.

Treatment	Time	N. of animals score 0	N. of animals score 1	N. of animals score 2	N. of animals score 3	% of sensitised animals
Negative control	24 h	5	0	0	0	0
Negative control	48 h	5	0	0	0	0
FEXO-07	24 h	10	0	0	0	0
FEXO-07	48 H	10	0	0	0	0

Table 2.8. Main study: summary of results of cutaneous reactions: positive control.

Treatment	Time	N. of animals score 0	N. of animals score 1	N. of animals score 2	N. of animals score 3	% of sensitised animals
Positive control	24 h	0	0	5	0	100
	48 h	0	1	4	0	100

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions adopted, the test item FEX0-07 is free of any sensitising capacity in the male guinea pig.

Executive summary:

The possible de1ayed sensitising capacity of FEX0-07 was evaluated in the male guinea pig in accordance with the General Requirements of OECD Guideline N. 406 and the method B6 of Commission Directive N. 96/54/EC. The experimental technique is based on these of Magnusson, Kligman, Guillot and Coll.

The application of the test item FEX0-07 did not induce colouring of the application site.

Determination of the degree of allergenicity at times 24 and 48 hours was based upon the percentage of animals in the group showing a reaction, rather than on the severity of the latter. Under the experimental conditions adopted, results obtained were as fol1ows: No irritation reaction was noted at times 24 and 48 hours in animals of the negative control group and in animals treated during the challenge phase with the test item FEX0-07 at the Maximum Nonirritant Concentration (MNJC).

Under the experimental conditions adopted, the test item FEX0-07 is considered free of any sensitising capacity in the male guinea pig.