## FINAL REPORT

Study Title



## SCREENING FOR THE EY E IRRITANCY POTENTIA L OF

EXP1313100, 10% (W/V ) IN PROPYLEN E GLYCOL USING THE

BOVINE CORNEAL OPACI TY AND PERMEABILITY TEST

(BCOP TEST)

Author

Test Facility

WIL Research Europe B.V

. Hambakenwetering 7

5231 DD ‘s

- Hertogenbosch

The Netherlands

Laboratory Project Identification

Project 507656

Substance 205622 /A

- Page 1 of 16

-EXP1313100 Project 507656

- Page 2 - 1. CONTENTS

1. CONTENTS

................................................................................................ ...................................... 2

2. STATEMENT OF GLP COM PLIANCE

............................................................................................. 3

3. QUALITY ASSURANCE ST ATEMENT

............................................................................................ 4

4. SUMMARY

................................................................................................ ........................................ 5

5. INTRODUCTION

.............................................................................................................................. 6

5.1. Preface

................................................................................................ ...................................... 6

5.2. Aim of the study

........................................................................................................................ 6

5.3. Guidelines

................................................................................................ ................................ . 6

5.4. Storage and retention of records and materials

........................................................................ 7

6. MATERIALS AND METHOD S

.......................................................................................................... 7

6.1. Test Substance

......................................................................................................................... 7

6.1.1. Test substance informatio n

............................................................................................. 7

6.1.2. Study specific test substance information

...................................................................... 7

6.1.3. Test substance preparation

............................................................................................ 7

6.2. Reference substances

.............................................................................................................. 7

6.3. Test system

............................................................................................................................... 8

6.4. Study design

............................................................................................................................. 8

6.4.1. Preparation of corneas

................................................................................................... 8

6.5. Cornea selection and Opacity reading

...................................................................................... 8

6.6. Treatment of corneas and opacity measurements

.................................................................... 8

6.7. Opacity measurement

............................................................................................................... 9

6.8. Application of sodium fluorescein

............................................................................................. 9

6.9. Permeability determinations

...................................................................................................... 9

6.10. Electronic data capture

........................................................................................................... 9

6.11. Interpretation

.......................................................................................................................... 9

6.11.1. In vitro irritancy score

................................................................................................ . 9

6.11.2. Acceptability of the assay

........................................................................................ 10

6.12. List of deviations

................................................................................................................... 10

6.12.1. List of protocol deviations

........................................................................................ 10

6.12.2. List of standard operating procedures deviation s

.................................................... 10

7. RESULTS

................................................................................................ ....................................... 10

8. DISCUSSION AND CONCL USION

................................................................................................ 11

9. REFERENCES

............................................................................................................................... 11

TABLES

Table 1 Summary of opacity, permeability and in vitro scores

.................................................. 12

Table 2 Opacity score

................................................................................................................ 13

Table 3 Permeability score individual values (uncorrected)

...................................................... 13

Table 4 Permeability score individual values (corrected)

.......................................................... 14

Table 5 In Vitro irritancy score

................................................................................................... 14

Table 6 Historical control data for the BCOP studies

................................................................ 15

APPENDICES

APPENDIX 1 SUMMARY TABLE

....................................................................................................... 12

APPENDIX 2 INDIVIDUAL OPACITY, PERMEABILITY AND IN VITRO SCORES

........................... 13

APPENDIX 3 HISTORICAL CONTROL DATA

................................................................................... 15

APPENDIX 4 CERTIFICATE OF ANALYSIS

...................................................................................... 16EXP1313100 Project 507656

- Page 5 - 4. SUMMARY

Screening for the eye irritancy potential of EXP1313100 , 10% (w/v) in propylene glycol using the

Bovine Corneal Opacity and Permeability test (BCOP test).

Thi s report describes the ocular irritation properties of EXP1313100 on an isolated bovine cornea. The

possible ocular irritancy of EXP1313100 was tested through topical application for 10 ± 1 minutes.

The study procedures described in this report were based on the most recent OECD and EC

guideline s.

Batch E00873

- 025

- 001 of EXP1313100 was a brown solid

. The test substance was applied as a

1 0% (w/v) suspension in p ropylene glycol directly on top of the corneas.

The negative control responses for opacity and permeability were less than the upper limits of the

laboratory historical range indicating that the negative control did not induce irritancy on the corneas.

The mean in vitro irritancy score of the solvent control (Propylene glycol) was 1.0 indicating tha t the

solvent control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive

control (10% ( w/v ) Benzalkonium Chloride) was 111 and was within the historical positive control data

range

. It was therefore concluded that the test conditions were adequate and that the test system

functioned properly.

EXP1313100 induced ocular irritation through only endpoint opacity , resulting in a mean in vitro

irritancy score of 4.1 after 10 minutes of treatment

.

Since EXP1313100 , 10% (w/v) in propylene glycol induced an IVIS > 3 ≤ 55, no prediction on the

classification can be made

.