N-(aminosulfonyl)-3-chloropropanimidamide hydrochloride

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

N-(aminosulfonyl)-3-chloropropanimidamide hydrochloride

EC Number:

700-156-6

Cas Number:

106649-95-0

Molecular formula:

C3H8ClN3O2S.HCl

IUPAC Name:

N-(aminosulfonyl)-3-chloropropanimidamide hydrochloride

Test material form:

solid: crystalline

Details on test material:

Test item: Chlorosulfamidine hydrochlorideCAS No.: 106649-95-0Batch No.: K41003NPhysical state: solid, crystalline substanceColour: whiteOdour: characteristicStorage: at a temperature of 15 to 30 °C

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

Species and strain: CBA/Ca Ola Hsd miceSource: TOXI-COOP ZRT.Hygienic level at arrival: SPFHygienic level during the study: Good conventionalNumber of animals: 24 animals/main test (4 animals/treatment group)Sex: Female, nulliparous, non-pregnantAge of animals: Young adult mice; 10-11 weeks old (at start of the main test)Body weight range at starting: 17.6-22.7 gAcclimatization time: 7 daysAnimal health: Only healthy animals were usedHousing during acclimatization period: Grouped caging in small groupsHousing during the test: Grouped caging (4 animals/cage)Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.Temperature: 22 ± 3 °CRelative humidity: 30 – 70 %

Study design: in vivo (LLNA)

Vehicle:

other: aqueous 1 % Pluronic; AOO (acetone:olive oil, 4:1) for the positive control groups

Concentration:

The test item was administered at three different concentrations (75; 50; 25 w/v%) according to the results of the dose range finding test.

No. of animals per dose:

4 animal/dose

Details on study design:

Selection of test item concentrations was based on the results of a formulation evaluation and also results of preliminary irritation/toxicity test to find an appropriate solvent and the maximum applicable concentration according to the relevant guidelines [1-2]. Based on the preliminary test results the test item was examined in the main test at 75 %, 50 % and 25 % (w/v) concentrations as formulations in the selected vehicle of aqueous 1 % (w/v) Pluronic®PE 9200 (aqueous 1 % Pluronic). Appropriate positive control (α-Hexylcinnamaldehyde [HCA] in 25 % (w/v) concentration), furthermore two negative control groups dosed with the vehicles of the test and positive control groups, respectively, were employed.The positive control item (25 % (w/v) HCA in Acetone: Olive oil 4:1 (v/v) mixture, AOO) induced the appropriate stimulation over the control, thus confirming the validity of the assay.

Positive control substance(s):

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

Results and discussion

Positive control results:

The positive control group animals were treated with 25 % (w/v) HCA solution (dissolved in AOO) concurrent to the test item groups. No mortality, cutaneous reactions or signs of toxicity were observed in the positive control group.Significant lymphoproliferative response (SI ≥ 3) was noted for HCA (SI = 12.8). The results of the positive control item demonstrated appropriate performance of the test in accordance with the relevant guidelines and confirmed validity of the assay.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

5.9

Variability:

chlorosulfamidine hydrochloride 75 % in aqueous 1 % Pluronic

Results 2

Parameter:

SI

Value:

11.3

Variability:

chlorosulfamidine hydrochloride 50 % in aqueous 1 % Pluronic

Results 3

Parameter:

SI

Value:

5.8

Variability:

chlorosulfamidine hydrochloride 25 % in aqueous 1 % Pluronic

Cellular proliferation data / Observations:

The stimulation index (SI = the DPN of a treated (positive control or test item) group divided by the DPN of the respective negative control group) for each treatment group was also calculated. A stimulation index of 3 or greater is an indication of a positive result. When monotonic dose-response is seen, attempt is made to calculate EC3 (dose calculated to induce a stimulation index of 3) with the help of linear regression on dose-response relationship. All calculations were made by Microsoft Excel Software. (See Table1 for the values.

Any other information on results incl. tables

Test Group Name	Measured DPM/group	Group* DPM	DPN (DPM/Node)	Stimulation Index Values
Vehicle control for the positive control: AOO	7283	7258.0	907.3	1.0
Positive control: 25 % HCA in AOO	93261	93236.0	11654.5	12.8
chlorosulfamidine hydrochloride 75 % in aqueous 1 % Pluronic	17314	17289.0	2161.1	5.9
chlorosulfamidine hydrochloride 50 % in aqueous 1 % Pluronic	33261	33236.0	4154.5	11.3
chlorosulfamidine hydrochloride 25 % in aqueous 1 % Pluronic	17112	17087.0	2135.9	5.8
Vehicle control for the test item: aqueous 1 % Pluronic	2977	2952.0	369.0	1.0

 *Group DPM = measured DPMgroup- average DPMbackground

Applicant's summary and conclusion

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Remarks:

Migrated information

Conclusions:

In conclusion, under the conditions of the present Local Lymph Node Assay, Chlorosulfamidine hydrochloride (CAS No. 106649-95-0) tested at the maximum attainable concentration of 75 % (based on solubility) and at concentrations of 50 % and 25 % (w/v) as formulations in an appropriate vehicle (aqueous 1 % Pluronic) was shown to have skin sensitization potential. An EC3 value could not be calculated in this LLNA.Because the EC3 value can not be calculated, sub-category can not be determined in this LLNA. The test item should be classified into GHS Category 1.