N-[(4-{5-AMINO-4-CYANO-1-[(2S)-1,1,1-TRIFLUOROPROPAN-2-YL]-1H-PYRAZOL-3-YL}PHENYL)METHYL]-5-FLUORO-2-METHOXYBENZAMIDE

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 June 2020 to 3 August 2020

Reliability:

1 (reliable without restriction)

Data source

Reference

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-[(4-{5-AMINO-4-CYANO-1-[(2S)-1,1,1-TRIFLUOROPROPAN-2-YL]-1H-PYRAZOL-3-YL}PHENYL)METHYL]-5-FLUORO-2-METHOXYBENZAMIDE

EC Number:

953-182-7

Cas Number:

2101703-40-4

Molecular formula:

C22H19F4N5O2

IUPAC Name:

N-[(4-{5-AMINO-4-CYANO-1-[(2S)-1,1,1-TRIFLUOROPROPAN-2-YL]-1H-PYRAZOL-3-YL}PHENYL)METHYL]-5-FLUORO-2-METHOXYBENZAMIDE

Specific details on test material used for the study:

Lot Number: 60394-18-001
Physical Description: Light Brown powder
Receipt date: 8 June 2020
Storage: Room temperature

Test animals / tissue source

Species:

cattle

Strain:

not specified

Test system

Vehicle:

other: Deionised water

Controls:

yes, concurrent vehicle

Amount / concentration applied:

20% (w/v) dilution in sterile, deionized water

Duration of treatment / exposure:

Three corneas were incubated in the presence of the test article at 32 ± 1 °C for 4 hours. Three corneas were incubated in the presence of each control at 32 ± 1 °C for 4 hours. After removal of the test or control article from the corneas, a final opacity was determined.

Observation period (in vivo):

The corneas did not receive a postexposure incubation

Number of animals or in vitro replicates:

3

Details on study design:

Three corneas were incubated in the presence of the test article at 32 ± 1 °C for 4 hours. Three corneas were incubated in the presence of each control at 32 ± 1 °C for 4 hours.
The test article was dosed and rinsed from the corneas using the open chamber method. In the open chamber method, the anterior chamber windows were removed and the test article was applied directly onto the exposed epithelium of the corneas. After exposure, the test article was directly rinsed from the exposed corneas.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

ca. 1.4

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the in vitro score of 1.4, and the prediction model found in OECD test guideline 437, the test article is predicted to not require classification or labelling for ocular irritation (GHS No Category) and the test article is predicted to be a non-irritant.

Executive summary:

The Bovine Corneal Opacity and Permeability Assay (BCOP) was used to assess the potential ocular irritancy of the test article to isolated bovine corneas. The methods and procedures used in this assay were consistent with OECD Test Guideline 437: Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2017).

The test article was administered to the test system as a 20% (w/v) dilutionin sterile, deionized water. The positive control (20% (w/v) dilution of imidazole prepared in Complete Minimal Essential Medium (without phenol red)) and the negative control (sterile, deionized water) were tested concurrently. After vortexing, the test article dilution was not fully soluble, and formed a cloudy light brown non-viscous suspension with brown particles.

Three corneas were incubated in the presence of the test article at 32 ± 1 °C for 4 hours. Three corneas were incubated in the presence of each control at 32 ± 1 °C for 4 hours. After removal of the control articles from the corneas, a final opacity was determined (i.e., the corneas did not receive a postexposure incubation). Following the rinsing process, but prior to reading the opacity values for the
corneas treated with the test article, a bulb burnt out in the opacitometer. The corneas treated with test article were placed back into the incubator (for approximately 30 minutes) until another opacitometer was calibrated which was then used to measure the final opacity readings.  The test article was dosed and rinsed from the corneas using the open chamber method. In the open
chamber method, the anterior chamber windows were removed and the test article was applied directly onto the exposed epithelium of the corneas. After exposure, the test article was directly rinsed from the exposed corneas.

For regulatory purposes, the In Vitro Score (IVIS) cut-off values for identifying test chemicals as inducing serious eye damage (UN GHS Category 1) and test chemicals not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are found in the table below (OECD 437, adopted 9 October 2017). This guidance on categorization applies only to a test article evaluated using the appropriate standard protocols as described in OECD 437.

IVIS	UN GHS
</=3	No Category
>3;</=55	No prediction can be made
>55	Category 1

The following classification system was used:-

In Vitro Score

Up to 4=Non-irritant

>4-12 = Slight irritant

>12.1 to 25 =Mild irritant

25.1 to 55 = moderate irritant

55.1 and above = Severe irritant

Based on the in vitro score of 1.4, and the prediction model found in OECD test guideline 437, the test article is predicted to not require classification or labelling for ocular irritation (GHS No Category) and is predicted to be a non-irritant.