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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
In Vivo Study was conducted prior to the availability of the LLNA or other in-vitro test methods

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report date:
1997

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: Conducted in compliance with EEC Methods for the determination of toxicity. Annex. of Directive 92/69/EEC, Part B, Method B.6.
Version / remarks:
Skin sensitisation method used was the guinea-pig maximisation test described by MAGNUSSON, B, And KLIGMAN, A.M. (1970) Aliergic Contact Dermatitis in the Guinea-pig: identification of contact allergens„ Thormis, C,C.. Springfield, Illinois, I A
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was conducted prior to the ammendments to Annex VII and the availability of the LLNA

Test material

1
Chemical structure
Reference substance name:
4-[2-Ethoxy-5-(4-methyl-1-piperazinylsulfonyl)benzamido]-1-3-propyl-1H-pyrazole-5-carboxamide
EC Number:
639-263-7
Cas Number:
200575-15-1
Molecular formula:
C22H32N6O5S
IUPAC Name:
4-[2-Ethoxy-5-(4-methyl-1-piperazinylsulfonyl)benzamido]-1-3-propyl-1H-pyrazole-5-carboxamide
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
Off White Powder, 95% purity

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
The guinea-pigs were housed in groups of five in suspended metal cages with wire mesh floors. A vitamin C enriched guinea-pig diet (Harlan Teldad 9600 FD2 SQC) and drinking water were provided ad libitum. Hay was given weekly. Thee batch of diet used for the study was anal ysed for nutrients, possible contaminants or microorganisms, likely to be present in the diet,and which, if in excess, may have had an undesirable effect on the test system. Animal room temperature wa.s controlled within the range 17,0 to 28,5 °C and relative humidity within
the range 39 to 67%. These environmental parameters were recorded daily. Air exchange was maintained at approximately 15 air changes per hour and lighting was controlled by means of a time switch to give 12 hours of artificial light (0700 - 1900 hours) in each 24 hours period. Each animal was identified by ear tattoo number. Animal were observed daily for signs of ill health or toxicity, Body weight of each guinea pig in the main study was recorded on Day 1 and on the last day of observations to the challenge applications.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: 5% Acetone in Alembicol D
Concentration / amount:
1) Freund's complete adjuvant was diluted with an equal volume of water for irrigation
2) 5% w/v in 5% acetone in Alembicol D.
3) 5% w/v in a 30 :50 mixture of Freund's complete adjuvant and 5% acetone in Alembicol D.
Day(s)/duration:
7
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, open
Vehicle:
other: acetone
Concentration / amount:
70%
Day(s)/duration:
48 hours
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
Challenge
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
70% w/v and 35% w/v
Day(s)/duration:
24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
10 animals per treated dose and 5 animals for control
Details on study design:
Based on the results of the preliminary investigations, the following concentrations of UK-220,955 were selected:
- Induction iritradermal injection - 5% w/v in 5% acetone in Alembicol D
This was the maximum practical concentration for intradermal administration and caused irritation but did not adversely affect the animals.

- Induction topical application - 70% w/v in acetone
- Topical challenge - 70 and 35% w/v in acetone
From preliminary investigations 70% wlv in acetone was the maximum practical concentration and did not give rise to irritating effects.
Challenge controls:
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. The control were challenged topically as per the treated animals

Positive control substance(s):
yes
Remarks:
The sensitivity of the guinea.-pig strain used is checked periodically with known sensitisers hexyt cinnamic aldehyde (HCA). Benzocaine and 2-mercaptobenzothiazole

Results and discussion

Positive control results:
All animals exposed to the HCA and the MBT showed positive responses.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
35%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
70%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
30%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
70
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
70% Acetone only
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
35% Acetone
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
4-[2-Ethoxy-5-(4-methyl-1-piperazinylsulfonyl)benzamido]-1-3-propyl-1H-pyrazole-5-carboxamide did not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the ten test animals.
Executive summary:

This study was performed to assess the skin sensitisation potential of 4-[2-Ethoxy-5-(4-methyl-1-piperazinylsulfonyl)benzamido]-1-3-propyl-1H-pyrazole-5-carboxamide using the guinea-pig. The method followed was that described in: EEC Methods for the determination of toxicity, Annex of Directive 92/69/EEC Method B 6. Skin sensitisaiion. MAGNUSSON, B. and KLIGMAN_A.M. (1970). Based on the results of a preliminary study and in compliance with the guideline, the following dose levels were selected:

Intradennal injection 5 w/v in 5% A acetone in Alembieol D

Topical application: 70% w/v in acetone

Challenge application: 70 and 35% w/v in acetone

Ten test and five control guine.a.-pigs were used in this study. 4-[2-Ethoxy-5-(4-methyl-1-piperazinylsulfonyl)benzamido]-1-3-propyl-1H-pyrazole-5-carboxamidedid not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the ten test animals.

4-[2-Ethoxy-5-(4-methyl-1-piperazinylsulfonyl)benzamido]-1-3-propyl-1H-pyrazole-5-carboxamide does not require classification or labelling accordance with GHS