[No public or meaningful name is available]

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 20 Dec 2016 to 19 Jan 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 442B (Skin Sensitization: Local Lymph Node Assay: BrdU-ELISA)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA): BrdU-ELISA

Test material

Test material information

Constituent 1

Reference substance name:

potassium (trans-4-(methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino)-cyclohexyl)-methanesulfonate

Cas Number:

2124221-14-1

Molecular formula:

C14H19KN4O3S

IUPAC Name:

potassium (trans-4-(methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino)-cyclohexyl)-methanesulfonate

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Jackson Laboratories (Bar Harbor, ME) on 07 Dec 2016.
- Females (if applicable) nulliparous and non-pregnant: yes
- Microbiological status of animals, when known:
- Age at study initiation: The animals were born on 18 Oct 2016, ± 3 days.
- Weight at study initiation: The Day 1 body weight range for the preliminary dermal irritation screen
animals was 18.0 - 21.6 grams. The Day 1 body weight range for the main test animals was 18.5 -
23.7 grams.
- Housing: During the acclimation period the animals were housed two per cage in suspended wire-bo
ttom cages; during the study the animals were housed one per cage. Bedding was placed beneath the
cages and changed at least three times per week.
- Diet (e.g. ad libitum): Fresh PMI Rodent Chow (Diet No. 5001) were available ad libitum.
- Water (e.g. ad libitum): water were available ad libitum.
- Acclimation period: at least five days
- Indication of any skin lesions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): continuously monitored
- Humidity (%): continuously monitored
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

10%, 25% and 50% [w/v]

No. of animals per dose:

5 females

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility:
- Irritation: no
- Systemic toxicity: no
- Ear thickness measurements: None of the test article treatments resulted in increases in ear thi
ckness of 25% or more, and therefore the test article was not considered irritating.
- Erythema scores: No test article treatment produced erythema.

MAIN STUDY
The test article concentrations used in the main LLNA study were chosen such that the
maximum concentration tested was the highest doseable formulation of the test article in the vehicle,
while avoiding both overt systemic toxicity and excessive local dermal irritation. Concentrations of
10%, 25%, and 50% (w/v) were chosen.

ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: A test article is considered to have a positive response
if treatment results in a 1.6-fold or greater increase in the mean LNC proliferation (BrdU ELISA OD
values) relative to that obtained for the vehicle control.

TREATMENT PREPARATION AND ADMINISTRATION: Treatment was made by topical application
of the test article concentrations to the dorsum of each ear once daily for three consecutive days. The
test article was spread over the entire dorsal surface of the ear using a micropipette to deliver 25 μl/
ear.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

For each test group, the individual animal SI values along with the mean group SI and standard devia
tion were calculated, and ANOVA followed by the Students’ t-Test was run to statistically compare ea
ch test article dose group to the vehicle control group. Although specified in the test guidelines, these
calculations and results were not incorporated into the interpretation of the data. An SI value of 1.6 or
more is the sole determinant for a positive sensitization response.

Results and discussion

Positive control results:

25% HCA (Positive Control) result in SI value of 2.4, indicates a sensitizing response.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

0.5

Test group / Remarks:

10% w/v

Results 2

Key result

Parameter:

SI

Value:

0.8

Test group / Remarks:

25% w/v

Results 3

Key result

Parameter:

SI

Value:

0.5

Test group / Remarks:

50% w/v

Cellular proliferation data / Observations:

Body weight losses were noted but were not significant (less than 2 g).
All animals survived the in-life phase of the study.
Erythema Scores:
No test article treatment produced erythema.
Ear Thickness as a Measure of Dermal Irritation:
In the main test, none of the test concentrations tested resulted in excessive dermal irritation.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Topical application of test article at 10%, 25%, and 50% (w/v) in DMSO resulted in SI values less than
1.6. Therefore, this test article is not a dermal sensitizer in the Local Lymph Node Assay.

Executive summary:

The skin sensitisation study is performed with LLNA according to OECD Guideline 442B under GLP.
Topical application of test article at 10%, 25%, and 50% (w/v) in DMSO resulted in SI values less than
1.6. Therefore, this test article is not a dermal sensitizer in the Local Lymph Node Assay.