4-(2-{(E)-2-[(1E)-1-{(2E)-2-[1,1-dimethyl-3-(4-sulfobutyl)-1,3-dihydro-2H-benzo[e]indol-2-ylidene]ethylidene}-2-{methyl[(trifluoromethyl)sulfonyl]amino}-1H-inden-3-yl]vinyl}-1,1-dimethyl-1H-benzo[e]indolium-3-yl)butane-1-sulfonate compound with N,N-diethylethanamine (1:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Test guideline 3

Qualifier:

equivalent or similar to guideline

Guideline:

other: The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006.

Test guideline 4

Qualifier:

equivalent or similar to guideline

Guideline:

other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.

Test guideline 5

Qualifier:

equivalent or similar to guideline

Guideline:

other: Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.

Principles of method if other than guideline:

List of protocol deviations:
1. The isolated corneas were stored in a petri dish with cMEM at room temperature.
Evaluation: For technical reasons, during the examination phase of the eyes and the isolation of the corneas, these were stored at room temperature. The corneas mounted in the corneal holder were incubated for the minimum of 1 hour at 32 ± 1°C. This deviation had no effect on the results of the study.
2. The time, on which the incubation with sodium-fluorescein was ended, was not registered.
Evaluation: The incubation time should be 90 ± 5 minutes. The time of the next step, the permeability measurement was registered and was 110 minutes. Furthermore the values observed for the negative and positive control were within the historical control data ranges. Therefore, it can be concluded that the incubation with sodium-fluorescein was approximately 90 minutes and this deviation had no effect on the results of the study.
The study integrity was not adversely affected by the deviations.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-(2-{(E)-2-[(1E)-1-{(2E)-2-[1,1-dimethyl-3-(4-sulfobutyl)-1,3-dihydro-2H-benzo[e]indol-2-ylidene]ethylidene}-2-{methyl[(trifluoromethyl)sulfonyl]amino}-1H-inden-3-yl]vinyl}-1,1-dimethyl-1H-benzo[e]indolium-3-yl)butane-1-sulfonate compound with N,N-diethylethanamine (1:1)

EC Number:

940-889-0

Cas Number:

1770781-13-9

Molecular formula:

C51 H52 F3 N3 O8 S3 . C6 H15 N

IUPAC Name:

4-(2-{(E)-2-[(1E)-1-{(2E)-2-[1,1-dimethyl-3-(4-sulfobutyl)-1,3-dihydro-2H-benzo[e]indol-2-ylidene]ethylidene}-2-{methyl[(trifluoromethyl)sulfonyl]amino}-1H-inden-3-yl]vinyl}-1,1-dimethyl-1H-benzo[e]indolium-3-yl)butane-1-sulfonate compound with N,N-diethylethanamine (1:1)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

97.85 % purity, green-brown powder, 0.87% water

Test animals / tissue source

Species:

other: bovine eye after slaughter

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Bovine eyes were used as soon as possible after slaughter on the same day.

Test system

Vehicle:

not specified

Controls:

yes

Amount / concentration applied:

CH03938 was weighed in a bottle and applied directly on the corneas in such a way that the cornea was completely covered (308 to 319 mg).

Duration of treatment / exposure:

240 +- 10 min

Observation period (in vivo):

/

Number of animals or in vitro replicates:

3 eyes as negative control, 3 eyes as positive control and 3 eyes tested on CH03938

Details on study design:

The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).
Additionally the opacity and permeability values were evaluated independently to determine whether the test substance induced irritation through only one of the two endpoints.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: mean opacity

Basis:

mean

Score:

29

Reversibility:

not specified

Results 2

Irritation parameter:

other: permeability

Basis:

mean

Score:

0.057

Reversibility:

not specified

Results 3

Irritation parameter:

other: in vitro irritation score

Basis:

mean

Score:

30

Reversibility:

not specified

Any other information on results incl. tables

Summary of opacity, permeability and in vitro scores

Treatment Mean Opacity Mean Permeability Mean In vitro Irritation Score 1, 2

Negative control 0 0.000 0.0

Positive control 104 1.315 124

CH03938 29 0.057 30

1 Calculated using the negative control mean opacity and mean permeability values.

2 In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

Applicant's summary and conclusion

Interpretation of results:

moderately irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Since CH03938 induced an IVIS > 3 ≤ 55.1, it is concluded that CH03938is mild to moderate irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.