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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 guidelineopen allclose all
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
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)
Qualifier:
equivalent or similar to guideline
Guideline:
other: The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006.
Qualifier:
equivalent or similar to guideline
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.
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

Constituent 1
Chemical structure
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

Resultsopen allclose all
Irritation parameter:
other: mean opacity
Basis:
mean
Score:
29
Reversibility:
not specified
Irritation parameter:
other: permeability
Basis:
mean
Score:
0.057
Reversibility:
not specified
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.