N-(N6-trifluoroacetyl-L-lysyl)-L-proline

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1989-11-14 to 1990-02-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1981

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-(N6-trifluoroacetyl-L-lysyl)-L-proline

EC Number:

402-920-1

EC Name:

N-(N6-trifluoroacetyl-L-lysyl)-L-proline

Cas Number:

103300-89-6

Molecular formula:

C13H20F3N3O4

IUPAC Name:

(2S)-1-[(2R)-2-amino-6-(2,2,2-trifluoroacetamido)hexanoyl]pyrrolidine-2-carboxylic acid

Test material form:

solid: particulate/powder

Test animals

Species:

rabbit

Strain:

other: White russian (Albino)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Males 9 months
- Weight at study initiation: Males 2.42 - 2.83 kg
- Housing: individually, Stainless steel cages with grating floor, type ASTA, size: 48.5x40x36.5 cm (LxBxH), supplied by ASTA Pharma AG, D-4800 Bielefeld 14
- Diet (e.g. ad libitum): Approx. 120 gram/day x animal Standard diet ssniff K, "Special diet for rabbits", supplied by ssniff Spezialfutter GmbH, D-4770 Soest
- Water (e.g. ad libitum): Water was provided ad libitum in drinking water quality from the Stadtwerke Bielefeld (Municipal Works). An automatic watering system with drinking nipples was used.
- Acclimation period: The animals were kept one day under test conditions before application of the product. Veterinary supervision of the animals was done before start of study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 - 22.5
- Humidity (%): 40-55
- Photoperiod (hrs dark / hrs light):12/12

Test system

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

other: moistened with demineralized water

Controls:

other: contralateral non-treated site at the same animal

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g moistened with 0.35 mL demineralized water

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.35 mL demineralized water

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48 and 72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: the shorn dorsal skin area between shoulder and sacrum.
- % coverage: (approx. 6.25 cm²)
- Type of wrap if used: (Acrylastic, No. 2408, width 8 cm; supplied by P. Beiersdorf and Co. AG, Hamburg

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the end of the exposition time remaining test material was gently washed off as far as possible.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1, 24, 48 and 72 hours after removal of the patches for 3 days p. appl.

SCORING SYSTEM: The evaluation of the cutaneous irritation effect at observation times 1, 24, 48 and 72 hours p. appl. was expressed by an irritation index
- Method of calculation: The arithmetical mean values of the individual grades obtained from the skin areas at observation times 1, 24, 48 and
72 hours were calculated - separately for erythema- /eschar and edema formation. The irritation index is represented by the sum of these two mean values.
A corresponding gradation was allocated to this index by the following scheme in a modified method according to Gilman et al..
Index Ranges Gradation
0.0-0.5 non-irritant
0.6-3.0 slightly irritant
3.1-5.0 moderately irritant
5.1-8.0 severely irritant

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

edema score

Basis:

animal: 1-3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The single application of 0.5 gram ε-trifluoroacetyl-L-lysyl-L-proline to the intact skins of three rabbits caused only very slight changes. Erythema occurred in two animals 24 hours after removal of the patch. The treated skin area showed a very slight erythema (grade 1). The finding had completely disappeared at day 3 of observation.The irritation index is 0.3. ε-trifluoroacetyl-L-lysyl-L-proline therefore is classified as non-irritant in this test system.

Other effects:

None

Any other information on results incl. tables

Reaction of the exposed skin	hours after exposure	DRAIZE grade
		Animal number
Erythema and Eschar formation:
	1	0	0	0
	24	0	1	1
	48	0	1	1
	72	0	0	0
Edema
	1	0	0	0
	24	0	0	0
	48	0	0	0
	72	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was tested for skin irritation in a study conducted according to OECD guideline 404 (1981). Erythema occurred in two animals 24 hours after removal of the patch. The treated skin area showed a very slight erythema (grade 1). The finding had completely disappeared at day 3 of observation. Systemic-toxic effects did not occur after dermal application of the test substance. The general condition of the test animals was not affected. Therefore, ε-trifluoroacetyl-L-lysyl-L-proline is classified as non-irritant in this test system according to Regulation (EC) No1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).