1-(3-chloropyridin-2-yl)-N-[4-cyano-2-methyl-6-(methylcarbamoyl)phenyl]-3-{[5-(trifluoromethyl)-2H-tetrazol-2-yl]methyl}-1H-pyrazole-5-carboxamide

Endpoint summary

Description of key information

Skin irritation (OECD 404), rabbit: not irritating

Eye irritation (OECD 405), rabbit: not irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 Apr - 06 Apr 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted in 2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

adopted in 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2008

GLP compliance:

yes (incl. QA statement)

Remarks:

GYEMSZI, National Institute for Quality- and Organizational Development in H ealthcare and Medicine, Budapest, Hungary

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: S&K-Lap Co.Ltd., Kartal, Hungary
- Age at study initiation: 13 - 14 weeks
- Housing: individually in AAALAC approved metal wire rabbit cages
- Diet: UNI diet for rabbits (Agribands Europe Hungary PLC, Karcag, Hungary), ad libitum
- Water: tap water from the municipal supply, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16.8 - 20.9
- Humidity (%): 24 - 56
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

approximately 24 h before application

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

Duration of treatment / exposure:

Initially, a single animal was treated for 1 h. As no significant irritant effect was observed after the 1 h exposure, the test was completed using the 2 remaining animals with an exposure period of 4 hours.

Observation period:

72 h
Reading time points: 1, 24, 48 and 72 h

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: 10 x 10 cm²
- Type of wrap: the test material was held in contact with the skin by gauze pads dampened with water to ensure good contact with the skin. An adhesive hypoallergenic plaster kept the gauze pads in contact with the skin. The entire trunk of the animals was then wrapped with plastic wrap with an elastic stocking.

REMOVAL OF TEST SUBSTANCE
- Washing: residual test material was removed with lukewarm water
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1, 24, 48, 72 h

SCORING SYSTEM:
- Method of calculation: numerical scoring system listed in the Commission Directive 2004/73/EC 2004 which is based on the Draize scoring system

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

edema score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

Very slight erythema (score 1) was noted in all animals at 1 h after patch removal and very slight erythema (score 1) was noted in two rabbits 24 hours after patch removal.

Other effects:

- Other adverse local effects: At the observation time points, 48 and 72 h after patch removal, there were no local signs on the skin of the treated animals.
- Other adverse systemic effects: No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred. The body weights of all rabbits were considered to be within the normal range of variability.

Interpretation of results:

other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 - 28 May 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted in 2012

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

adopted in 1998

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

adopted in 2008

GLP compliance:

yes (incl. QA statement)

Remarks:

GYEMSZI National Institute for Quality- and Organizational Development in Healthcare and Medicines, Budapest, Hungary

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: S&K-LAP Kft., Kartal, Hungary
- Age at study initiation: 10 - 11 weeks
- Weight at study initiation: 2765 - 2803 g
- Housing: individual in AAALAC approved metal wire rabbit cages
- Diet: diet for rabbits (Agribands Europe Hungary PLC, Karcag, Hungary), ad libitum
- Water: municipal tap water, ad libitum
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.1 - 20.1
- Humidity (%): 40 - 75
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g

Duration of treatment / exposure:

1 h

Observation period (in vivo):

72 h
Reading time points: 1, 24, 48 and 72 h

Number of animals or in vitro replicates:

3 males

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: The treated eye was rinsed with physiological saline solution.
- Time after start of exposure: 1 h

SCORING SYSTEM: Draize scoring system and OECD 405

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: reversibility: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Other effects:

- Lesions and clinical observations: There were no mortalities and no clinical signs of systemic toxicity observed during the study. The general state and behaviour of animals were normal throughout the study period.
- Ophthalmoscopic findings: Fluorescein staining was negative in all animals during the observation period.
- Other observations: Discharge was noted 1 h post application in 3/3 males but was reversible within 24 h. The body weight and body weight changes were considered to be normal with no indication of any treatment related effect.

Table 1 Individual mean scores

Animal number	Effects	24 h	48 h	72 h	Mean scores	Response	Reversible (days)
1	Corneal opacity Iritis Redness conjunctivae Chemosis conjunctivae Discharge	0 0 1 1 0	0 0 1 0 0	0 0 0 0 0	0.00 0.00 0.67 0.33 0.00	- - - - -	na na 3 2 1*
2	Corneal opacity Iritis Redness conjunctivae Chemosis conjunctivae Discharge	0 0 1 0 0	0 0 0 0 0	0 0 0 0 0	0.00 0.00 0.33 0.00 0.00	- - - - -	na na 2 1* 1*
3	Corneal opacity Iritis Redness conjunctivae Chemosis conjunctivae Discharge	0 0 1 0 0	0 0 0 0 0	0 0 0 0 0	0.00 0.00 0.33 0.00 0.00	- - - - -	na na 2 1* 1*

-- = negative, na = not applicable, * = in respect of the result 1 h post application

Interpretation of results:

other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Reliable studies regarding skin irritation/ corrosion and eye irritation/ corrosion are available for the test substance.

Skin:

The acute skin irritation potential of the test substance was assessed in a study performed according to OECD Guideline 404 and in compliance with GLP (M-462513-01-1). Three male young adult New Zealand White rabbits were treated by topical semi-occlusive application of 0.5 g of the test item to the intact shaved flank for 4 h. The treated skin surface was examined at 1, 24, 48 and 72 hours after patch removal. Irritation was scored by the method of Draize as described in the OECD guideline 404. Very slight erythema (score 1) was noted in all animals at 1 hour after patch removal and very slight erythema (score 1) was noted in two rabbits 24 h after patch removal. The mean scores for the 24, 48 and 72 h reading for erythema were 0.33 for 2/3 and 0.0 for 1/3 animal. The mean score for the 24, 48 and 72 h reading for edema for each animal was 0, respectively. No mortality occurred and no clinical signs of systemic toxicity were observed during the study. The body weights of all rabbits were considered to be within the normal range of variability. Thus, under the conditions of this study, the test substance is a mild irritant to the skin of male New Zealand White rabbits. The very slight erythema observed in all animals was reversible within 48 h post-treatment. The results of the study do not trigger a classification of the test substance according to Regulation (EC) No 1272/2008 and thus, the test substance is considered to be not irritant to the skin.

Eye:

The acute eye irritation potential of the test substance was assessed in a study performed according to OECD Guideline 405 and in compliance with GLP (M-462510-01-1). 0.1 g of the test substance was placed into the conjunctival sac of the left eye of three male young adult New Zealand White rabbits. The eyes were washed 1 h after application and examined 1, 24, 48 and 72 h after application. The untreated right eye of each animal served as control. Fluorescein staining was performed 24 h before administration and 24, 48 and 72 h after application. One hour after the application, conjunctival redness (score 2), chemosis (score 2 or 1) and discharge (score 2) was noted in all rabbits. At 24 h after treatment, conjunctival redness (score 1) was noted in all animals and chemosis (score 1) was noted in 1/3 rabbit. At 48 h after treatment, conjunctival redness (score 1) was noted in 1/3 animal. At 72 h after treatment, no clinical signs, and no conjunctival or corneal effects were observed. The mean scores for the 24, 48 and 72 h reading for conjunctivae were 0.67 for 1/3 and 0.33 for 2/3 animals. The mean chemosis scores for the 24, 48 and 72 h reading were 0.33 for 1/3 and 0 for 2/3 animals. The mean scores for the 24, 48 and 72 h reading for cornea opacity and iris for each animal were 0, respectively. Fluorescein staining was negative in all animals during the observation period. There were no notable body weight changes during the study period. Thus, under the conditions of this study, the test substance caused conjunctival irritant effects and chemosis in the eye of male New Zealand White rabbits. The observed effects were fully reversible within 72 h post-treatment in all animals. The results of the study do not trigger a classification of the test substance according to Regulation (EC) No 1272/2008 and thus, the test substance is considered to be not irritant to the eyes.

Justification for classification or non-classification

The available data on skin and eye irritation do not meet the criteria for classification according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.