Benzoxazole, 5-chloro-2-[(5R)-hexahydro-5-methyl-1H-1,4-diazepin-1-yl]-, hydrochloride (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)

Rationale for reliability incl. deficiencies:

other: study performed according to standard in vitro methods; GLP-compliant (EPA, FDA)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Standard methods cited in literature.

Principles of method if other than guideline:

Protocol based on methodology described in ‘Bovine Corneal Opacity and Permeability Test: An In Vitro Assay of Ocular Irritancy (1982)’; Gautheron, P., Dukic, M., Alix, D., and Sina, J.F.; Fundamental and Applied Toxicology, 18, 442-449. In Vitro classification based on Southee, J.A., 1998. Evaluation of the Prevalidation Process, Part 2, final report, volume 2, The Bovine Corneal Opacity and Permeability (BCOP) Assay.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(7R)-4-(5-chloro-1,3-benzoxazol-2-yl)-7-methyl-1,4-diazepan-1-ium chloride

EC Number:

690-742-7

Cas Number:

1266664-66-7

Molecular formula:

C13H17Cl2N3O

IUPAC Name:

(7R)-4-(5-chloro-1,3-benzoxazol-2-yl)-7-methyl-1,4-diazepan-1-ium chloride

Details on test material:

Purity > 98 %

Test system

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): 20% solution in MEM (Minimum Essential Medium)
VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

Details on study design:

As per quoted methods.
In brief, five bovine corneas were 'dosed' with 0.75 mL of a 20% solution (in MEM, Minimum Essential Medium) of the test item. Opacity measurements and sodium fluorescein permeability were determined.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

ca. 119

Other effects / acceptance of results:

The corrected mean optical score was 118.2. The corrected mean optical density (permeability) score
was 0.054. The in vitro score in this BCOP study was calculated as 119.0 and, accordingly, the test
item was classified as a very severe irritant.

Any other information on results incl. tables

Results

Individual Control Scores for BCOP

Cornea #:	Pretest	4 Hours	O.D. Scores
C3	0	0	0.043
C4	0	0	0.019
Mean	0	0	0.031
Corrected Mean Control Opacity Score	0

Corrected Mean Control Opacity Score = 4 hour mean score minus pretest mean score

Individual Test Scores

Cornea	Pretest Scores				4 Hour Scores				O.D. Scores
6	C3	-1	C4	-2	C3	122	C4	122	0.053
7	C3	-1	C4	-1	C3	129	C4	128	0.056
8	C3	-2	C4	-2	C3	94	C4	92	0.103
9	C3	-2	C4	-1	C3	112	C4	112	0.124
10	C3	-4	C4	-4	C3	125	C4	125	0.087

Calculated Scores

Cornea #	Corrected Opacity Scores				Corrected O.D.
4 Hour Scores
6	C3	123	C4	124	0.022
7	C3	130	C4	129	0.025
8	C3	96	C4	94	0.072
9	C3	114	C4	114	0.093
10	C3	129	C4	129	0.056
Corrected Mean Optical Density =					0.054
Corrected Mean Optical Score =					118.2

Corrected Mean Opacity Score (CMOS) = mean treated opacity score minus corrected mean control opacity score Calculated In Vitro Score

CMOS =118.2 + 15 (0.054)

CMOS =118.2 + (0.810)

CMOS =119.0

Applicant's summary and conclusion

Interpretation of results:

other: highly irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

The in vitro score in this BCOP study was calculated as 119.0 and, accordingly, the test item was classified as a very severe irritant.

Executive summary:

This study protocol is based on methodology described in ‘Bovine Corneal Opacity and Permeability Test: An In Vitro Assay of Ocular Irritancy (1982)’. Five bovine corneas were 'dosed' with 0.75 mL of a 20% solution (in MEM, Minimum Essential Medium) of the test item. Opacity measurements and sodium fluorescein permiability were determined. The in vitro score in this BCOP study was calculated as 119.0 and, accordingly, the test item was classified as a very severe irritant.