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Toxicological information

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:
4.12.1978 - 19.01.1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1979
Report date:
1979

Materials and methods

Principles of method if other than guideline:
- Principle of test: Investigation on sensitizing potential of the test substance. During an induction phase the test substance as well as a positive and a negative control were intracutaneously applied for 10 times within 3 weeks. Freund's adjuvans was mixed to the test solutions in week 2 and 3. 13 days after the last induction dose, a single challenge dose was applied intradermally. A second challenge dose was applied epidermally 12 days thereafter (maximum subirritant dose). Animals were investigated for positive reactions.


- Parameters analysed / observed: skin thickness and the two largest diameter of reddend skin parts before treatment and 24 hours after treatment were determined during week one of induction phase and at the 1. challenge dose; a reaction volume [µl] was calculated based on thickness and diameter; values of week 1 (induction phase; local tolerance) were compared to value after 1. challenge; following the 2. challenge dose the reddening of the skin was taken as an allergic reaction.
GLP compliance:
not specified
Type of study:
Maurer optimisation test
Justification for non-LLNA method:
the guinea pig optimisation test was an adequate in vivo skin sensitisation test at the time of performance in 1979

Test material

Constituent 1
Chemical structure
Reference substance name:
Gestonorone caproate
EC Number:
215-010-2
EC Name:
Gestonorone caproate
Cas Number:
1253-28-7
Molecular formula:
C26H38O4
IUPAC Name:
17-acetyl-3-oxoestr-4-en-17-yl hexanoate

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Züchter Hagemann
- Weight at study initiation: 302 - 482 g
- Diet: ad libitum
- Water: ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 - 22.5
- Humidity (%): 35 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: Benzyl benzoate, castor oil
Concentration / amount:
0.1 mL (10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil)
Day(s)/duration:
Day 1, 3, 5
Adequacy of induction:
not specified
Route:
intradermal
Vehicle:
other: benzyl benzoate, castor oil
Concentration / amount:
2 mL (200 mg gestonorone caproate, 1218.8 mg benzyl benzoate, 700 mg castor oil) + 2 mL Freund's adjuvant
Day(s)/duration:
Day 8, 10, 12, 15, 17, 19
Adequacy of induction:
not specified
Challengeopen allclose all
No.:
#1
Route:
intradermal
Vehicle:
other: benzyl benzoate, castor oil
Concentration / amount:
0.1 mL (10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil)
Day(s)/duration:
13 days after last induction dose (day 32)
Adequacy of challenge:
not specified
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
other: benzyl benzoate, castor oil
Concentration / amount:
0.1 mL (10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil)
Day(s)/duration:
day 44
Adequacy of challenge:
other: local tolerable
No. of animals per dose:
20 (10 males/10 females)
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 9
- Exposure period: 3 weeks
- Test groups: Depostat I (0.1 mL [10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil]), Depostat II (same formulation, but artificially aged; increase in peroxid value from 1.4 to 8.2)
- Control group: Negative control: 0.9% (w/v) NaCl (day 1, 3, 5); 2 mL 0.9% (w/v) NaCl + 2 mL Freund's adjuvant (day 8, 10, 12, 15, 17, 19)
Positive control: 0.1% (w/v) DNCB
- Site: back and lateral thoracic wall
- Frequency of applications: 2-3 days
- Concentrations: 10 % (w/v)

B. CHALLENGE EXPOSURE
Challenge #1
- No. of exposures: 1
- Day(s) of challenge: day 32
- Test groups: Depostat I (0.1 mL [10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil]), Depostat II (same formulation, but artificially aged; increase in peroxid value from 1.4 to 8.2)
- Control group: Negative control: 0.9% (w/v) NaCl
Positive control: 0.1% (w/v) DNCB
- Site: back and lateral thoracic wall
- Concentrations: 10 % (w/v)
- Evaluation (hr after challenge): 24
Challenge #2
- No. of exposures: 1
- Day(s) of challenge: day 44
- Test groups: Depostat I (0.1 mL [10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil]), Depostat II (same formulation, but artificially aged; increase in peroxid value from 1.4 to 8.2)
- Control group: Negative control: 0.9% (w/v) NaCl
Positive control: 25% (w/v) DNCB
- Site: occlusive tape (2x2 cm)
- Duration: 24 hours
- Concentrations: 10 % (w/v)
- Evaluation (hr after challenge): 24 after removal of occlusive tape


OTHER:
2 treatment groups: Depostat I, Depostat II
negative control
positive control
deviation from induction protocol: Depostat II - 2.5 ml Depostat II + 2.5 ml Freund's adjuvant were applied at day 8,10, 12, 15, 17, 19

Positive control substance(s):
yes
Remarks:
2,4-Dinitrochlorobenzene (DNCB)

Results and discussion

Positive control results:
sensitizing

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0.9% NaCl
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0.9% NaCl
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.1% DNCB
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
positive control
Dose level:
25% DNCB
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10% (Depostat I)
No. with + reactions:
5
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
other: local intolerability
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
10% (Depostat I)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
other: Depostat II (artificially aged)
Dose level:
10%
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
other: local intolerability
Reading:
rechallenge
Hours after challenge:
24
Group:
other: Depostat II (artificially aged)
Dose level:
10%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
not specified
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

The observed positive reactions after the first challenge in the test item group Depostat I (5 of 20 animals) and in the additional artificially aged test item group Depostat II (3 of 20 animals) were not regarded as a sensitising effect because in both groups local intolerances were observed following intracutaneous treatment. The differences of the reaction volumes are slight compared to the ones during induction phase. Further, a sensitising effect could not be observed following a second challenge dose.

Applicant's summary and conclusion

Interpretation of results:
other: not sensitising
Executive summary:

The test item gestonorone caproate (formulated as 0.1 ml Depostat - 10 mg gestonorone caproate, 60.94 mg benzyl benzoate, 35 mg castor oil) did not show sensitising potential in an in vivo guinea pig optimisation test. The validity of the test procedure was confirmed by positive and negative controls.