### FINAL REPORT



### STUDY TITLE ACUTE EYE IRRITATION STUDY IN RABBITS STUDY No.: 

### DATA REQUIREMENT U.S. EPA Health Effects Test Gu id e lin es , OPPTS 870.2400 (1998) OECD Gu id el in es for the Te s ting of Chemica ls, Test No
. 405 (2002) JMAFF 12-No u s an-8147 (2000) Official Journal ofthe European Commun iti es, Met h ods for the Determination of Toxicity and Other Health Effe ct s, Part B .5 (Eye Irritation) ,

EYE Copy No. 212 

### Counci I Re g ul a tion (EC) No
. 

### 440/ 2004 

### AUTHOR

STUDY COMPLETED O N 16 June 2011

SPONSOR Dow Europe GmbH Bachtobel s tra sse 3 CH-88 

### J 0 Horgen Sw itzer l and 

### TEST FACILITY D E PARTMENT OF SAFETY ASSESSMENT ADVINUS TH ERA PEU TICS LIMITED POST BOX No. 58 1 3, PLOT Nos
. 21 & 

### 22 PE ENY A II 

### PHAS E, BANGALORE
- 560 058 I NDIA Page 1150

### COMPLIANCE WITH GOOD LABORATORY STANDARDS Test S ub s ta nce: ( Study Tit le: Ac ut e Eye Irritation Stud y in Ra bbit s Exce pt as not e d b e low, the st ud y described in thi s report was conducted in compl ia n ce w ith the following Good Labora tor y Practice Sta nd a rd s : Organi sa tion for E conomic Co -op e ration and Dev e lopment (O EC D) ENV / MC I CHEM (98) 1 7 European Eco nomic Community ( EEC) Comm i ss ion Directive L50/ 44 , 

### Feb.20, 2004 Jap a ne se M ini st r y ofAgricultu re , Forestry , and Fi s herie s (JMAFF) No u sa n No tificati o n No
. 6283 Agr icultural Production Bu reau I Oct o ber , 1999
Exc e ption to GLP compliance i s as li s ted be low: • Arch iv in g of raw data at the te st fac ilit y . Exce ption to Test guide lin e is as li ste d b e low:
• Exce ption to guideline 405 , for not te st in g we ig ht of evid e nce analys is by th e te st facility
.

Study Director

EYE Copy No. 212 

### 1 6 

## 1."",,**~** 

### 1\ 

### Date

Page 2 /5 0

### QUALITY ASSURANCE STATEMENT The Study No
. : entit led " Acute Eye Irrit ation Study in Rabbits " has been inspected in accordance w ith the OECD Principles of Good Laboratory Practice (as revised in 1 997) ENV /

MC I CHEM 

### (98)17 , OECD Paris, 1998, JMAFF GLP standards: II-Nousan-No.6283, I October 1999, and EC Directive 200411 O /

EC , 

### Official Journal of the European Union , L50/ 44 , Feb.20 , 2004. This study was in spected and findings reported to Management and to the Study Director on the dates shown below: INSPECTION DATE
28 February 20 I I

14 March 20 1 I

25 Apr il 20 I I and 26 Apr il 20 II 06 June 201 I PHASE Initiation Phase Review of study plan In-Life Phase Test substance in stillation (s in gle animal test) Reporting Phase Draft report review

Final report review REPORTING DATE
28 February 20 II

21 March 20 1 I

26 Apr il 201 I

06 June 20 I I Inspections were performed accordi ng to the Standard Operating Proced ur es of the test fac ilit y's Quality Assurance Un it. The final report was in spected aga in st the approved study plan and pertinent raw data and accurately reflects the raw data.

Date: If, .Jv ",< , 'VOll

EYE Copy No. 2/ 2 

### Head, GLP Section Quality Assurance Unit Advinus Therapeutics Limited Bangalore

Page 3/50

### 1. STUDY PERSONNEL T h e followin g per s on ne l participated in the conduct ofthe s tud y. Na me , Responsibility, Sect ion / Department Study Director Toxico logy Department of Safety Asse ss ment
Technica l Co
- ordinator Tox icology Department of Safety Assess ment

Cl ini cal Veterinarian Toxicology Dep a ltm ent of Safety As s e ss ment
Technical Assistance Toxicology Department of Safety A ss e ss ment
Document preparat ion Toxicology Department of Safety Assessme nt

EYE Co p y No. 2/ 2 

### F unction S ignatu r e with date
. Overall in-charge for the co nduct of the s tudy and report preparation A ss is tance in conduct of s t udy and report preparat ion
Veterinar y se rvic es.

Ass ista nce in the conduct of st udy

Data e ntr y and repo r t comp il atio n

P age 4/50

### TABLE OF CONTENTS STUDY TITLE
.. ..................................................... . .... .. .... .. ... ... ... ... ... . ...... .. . ..... . .. ... ...... 1 COMPLIANCE WITH GOOD LABORATORY STANDARDS
...... .. ...... . .. ............... 2 QUALITY ASSURANCE STATEMENT
.. . .. . .. ... . .. ... ...... ... ... ... .... .. ... . .. . ..... .. ... . ... . .. . . . ... 3 1. 

### 2. 3. 4. 5. 6. 

### 7. 8. 8
. 1 8
. 1.1 8.1.2 8.2 8.2.1.1 8.2.2 8.2
. 3 8.2.4 8
. 3 8.3.1 8.3.2 8.3.3 9. STUDY PERSONNEL
... .. . ....... . . .... ............... . ................... . .......... . ......... ...... 4 SUMMARy
......................... . .. . .... . . . . .. .......... . ... . . . .............. . .. . ........ ... ............. 7 STU DY DETAIL S
..................... . .. . ... . ................ . . . .... . ......... . . . .. ........ ..... . .. .. ... 8 OBJECTIVE
....................................................... . ........ .. ......................... . .. ... 9 JUSTIFI CAT IO N FOR SELECTION OF TEST SYSTEM
. .. .. . . . . .. . . . . .. . . .. ... 9 J UST IFICATIO N FOR ROUTE OF ADMlNISTRATION
. .. . . .. . ........... . ..... 9 TEST PRO CEDURE GUIDELINES
.. .. . .. . .. ..................... . ..... ... .. ......... . ....... 9 MATERIALS AND METHODS
. .. . .. .. . ...... .. .......... ... ... ... .... . . .. . . .................... 9 Materia l s ................... . . . .. . ... . ............................ . .. .. .. ....................................... 9 Test substance Information
. . ............ . .... ...... .. .... . . ... ......................... .. ............ 9 Test System
. ........ ...... . ............. . . . ... . .. . .. . ... .. . ... . ...... .. ...... ... .. . . .. ... .. ................. II Methods
.. . ... .. ....... ... ........... . .. ... ...... . .. . .. . ............................................ ... ........ II Hu sba ndry . . ................................. . ......... .. ...... . ... .. ............ . .. .. . . ................ . .... II Test Substance In st illation
. .. . . . . . .. . . . .. . .. . .................... . ... ............ ......... . . . . ...... 12 Animal Re st rain t
. .. ... . ...... .. . .. . . .... . ............. .. .••............................•................. 1 3 Stepwise exposure ofAnimals
. ........... . ...... .... ..... .. ... . ...... .... .. . ... .. . .... . .......... 1 3 Observation s ............. . .. ... . .. . .... . . . . .. . ................ ... .... ............ . ......................... 13 Toxic Signs and Pre-Terminal Deaths
....... ... . . ..... . ............... ... . . .. . . . .. . ..... . .... 13 Body Weight s .... .. ... .. . ... .. . .. . .. . . . . . . . . .. . . .. . ............. . .. . .. ........... .. ........ . . . ... . ......... 1 3 Eye Scoring
.................. . .. . ...... ..... .. ...................... ..... . . .. .. . ...... . ............ . .. . ..... 1 3 RESULTS
............ . . ... ........... .. ...... .. ......... ...... .. .. .. .. . ... .. . ... .. . .. . . ... .... . ... . ..... . ... 14

·EYE Copy No. 2/2 Page 5/50

### 9. 1 Toxic s ign s and pre-terminal deaths
.................. .... ............... .. . . .................. 14 9.2 Body we ights
. .................. ......................... . ......... . .. .. .. .............. ... ................ 14 9.3 Eye reactio n s .... .. .......... . .. ... .. ................. .... ..... ........ ... ................ .. ................ 14 10. CONCLUSION
.................. . . . .... .. ............ ... ............. ... . . . ............... .. ............. 14 II. ARCH IVING
............................ . . ........... .... . .. ............. ... ............. ................. 14 1 2. REPORT DISTRIBUTION
........ .. ... ... ......... .. .............. .. ............... .... . ......... 1 5 13. TABLES
............................................. ........................................................ 16 TABLE I. Body Weight , Toxic S igns and Pre-terminal deaths
............. 17 TABLE 2. Individu a l Eye Irritation Scores
.......... .......................... .. ...... 18 TABLE 3. Mean Eye Irrit at ion Score s .................................................... 19 14. APPENDICES
............................................................................................ 20 APPEND IX I. Certificate of Analysis
.................................................... 2 1 APPENDIX 2. N um er ic a l Scoring System (Grades of Ocular lesio n s ) .22 APPEND IX 3. Study Plan
...................................................................... 24 APPENDIX 4. Deviation fro m the Approved Study Plan
........ . ............ .43 1 5. ANNEXURES
............................................................................... . . ........... 44 ANNEXURE I
. Ana lys i s Report
- Anima l Diet Samp le ............ .... ..... ... .45 ANNEXURE 2. Ana lys i s Report
- Water Samp le ................ ......... ..... ..... 46 ANNEXURE 3. GLP Cert ifi cat e - Germany
.................................. .. ...... .47 ANNEXURE 4. GLP Certificate
- The Net h erland s .......... .. ................... .48 ANNEXURE 5. GLP Cert ifi cate - India
.................................................. 50

EYE 

### Co p y No. 2/2 Page 6/50

### 2. SUMMARY The acute eye irrit at ion / corro s io n s tudy in rabbit s wa s conducted to eva lu ate the eye irr i t at io n p ote nti a l of U nd er the condit io n s of thi s s tudy , ca u s ed conjunctival irrit ation a nd co rn ea l opac it y in the ra bb i t e ye, w hi ch c lea red b y 72 h o ur s . O n e h undred mg (0.1 g) of the te s t s ub stance wa s in s ti ll ed i nto the co njun ctiva l s ac of the left e y e ofthe a nim al afte r ge ntl y pulling the lower li d away from the e y eba ll. T h e lid s were gently held together for abo ut one s econd, in order to minimi z e lo ss of the te s t s ub s tance
. The right eye of eac h rabbit rema in ed untr eated and s erved a s the reference control. The oc u lar le s ion s were eva lu ated at I , 24 , 48 and 72 hour s accord ing to th e sca le fo r s cor in g ocu la r lesion s . There we re no toxic s ig n s or animal death
. 

### All rabbi t s appea red h ealt hy and ga in ed bod y wei g ht throu g h the ob s e rvat ion period. Und e r the conditions of thi s s t ud y , the m ea n eye irrit at ion sco r e s we r e as fo ll ows:

Observations Mean score at p o s t Conj un ctiv a Iri s Co rn ea Tota l mean instillation s core
I hour 6.00 0 0 6.00 24 hour s 6
. 00 0 6.67 12.67 48 h our s 2
. 67 0 3.33 6.00 72 hour s 0 0 0 0
The indi vidua ll y determined m ean irrit at io n sco r es for each an im a l for co rn ea l o p ac ity , iri s lesion , conj un ctiva l rednes s a nd co njun ctiva l c h emosis for t h e 24, 48 and 72 -h our in terva l s a re :

EYE Co p y No . 2/ 2 

### A n ima l No
. (Sex) RB8767 (M) RB8768 (M) RB8769 (M) Corneal Opacity 0.7

0 0 Iri s Le s ion Conj un c t i va l Co n ju n ctiva l Redne ss C hemo s i s 0 1.3 0.7 0 0.3 0 0 0. 3 0
Pa ge 7/ 5 0

### 3. STUDY DETAILS Study Title
Test s ub s ta nc e Study N umber Sponsor

Monitoring Scientist

Test Facility

Study Schedule Study initiation date Experimental s tarting date Accl imati zatio n
Treatment Observations Expe rimental completion date Draft report Study completion date ·EYE Copy No. 212 

### Acute Eye Irritati o n Study in Rabbits. (

Dow E urope GmbH Bachtobelstrasse 3 CH-8810 Horgen Switzerland Donna Anderson The Dow Chemical Company Toxicology & 

### E nvironmental Research & 

### Consulting (TERC) Building 5 727 Norristown Road, P.O. Box 904 Spring Hou se, PA 19477-0904 Email: DonnaAnderson@dow.com Department of Safety Assessment Advinus Therapeutics Limited Post Box No. 58 I 3, Plot Nos. 21 & 

### 22 Peenya II Pha se, Bangalore 560 058, INDIA
07 March 201 I 09 March 2011 Start: 09 March 201 I End: 13 March 2011 (single animal) and 14 March 2011 (additional two animals) Start: 14 March 20 II (single animal) 15 March 2011 (additional animals) Start: 14 March 2011 En d: 18 March 2011 18 March 2011 25 May 2011 16June2011 Page 8/50

### 4. OBJECTIVE The objective of thi s acute eye irrit ation s tud y in rabbit s was to provide information on t he potential for irritation arising from a s ingle instillation of a te s t s ub sta nce into the eye. 

### 5. JUSTIFICA TION FOR SELECTION OF TEST SYSTEM The rabbit is the system of choice becau se, historicall y, it has been the preferred and most commonly u se d s pecie s for primary eye irrit ation test s . 

### 6. JUSTIFICATION FOR ROUTE OF ADMINISTRATION The oc ul ar route of administration was u se d becau se it is recommended in the referenced gu ide lin es and because human exposure may occur via this route. 

### 7. TEST PROCEDURE GUIDELINES 

# » U.S. EPA Health Effects Test Guidelines, OPPTS 870.2400 , Acute Eye Irritation , August 1998. 

# » OECD Guide lin e for Testing of Chem icals, Sect ion 4 , Number 405. "Ac ute Eye Irritation / Corrosion" adopted 24th April 2002. 

# » JMAFF 12-Nou sa n-8147 , November 2000. 

# » Officia l Journal of the European Determination of Toxicity , Part 2004/73/EC, 29 Apri l 2004
. Communities, Methods for the 8.5 (Eye Irritation) , Directive


### 8. MA TERIALS AND METHODS 

### 8.1 Materials 

### 8.1.1 Test substance Information (As furnished b y the Sponsor) Te s t s ub s tance / Item Lot No.

EYE Copy No. 212 Page 9/5 0

### Manufactured by (Na me and Ad dr ess) S uppli ed by (Na m e a nd Address) Date of manufacture D ate of exp iry Purit y as p er Ce rtific ate of Analysi s Ph ys ic a l a pp ea ran ce Sto rage condit ions The Dow Chem ic a l Co mp any Midland , MI 48674 USA
The Dow Che mi ca l Co mp any M id land, MI 48674 USA
Marc h 20 10
06 Marc h 20 12

Off
- w hi te so lid

: Ambie nt (+ 1 8 to + 36°C) T h e test substance, ide ntifi ed as was receiv ed o n 03 February 20 I I and was further id e ntifi ed as 008-2 6 b y test fac ili ty, Reference N umb er dated 07 February 20 II. The test s ub sta n ce i s a so lid and i s s tored at amb ie nt cond iti on (+ 18 to + 36 ° C)
. Documentation of the m et h ods of sy nthe s i s , fa br icat ion, o r d e rivation of the te s t s ubsta nce is retained b y the Sponso r. D eter min at ion of th e s tab ili ty , identit y, st re n gt h , puri ty a nd compos ition of th e t es t s ub s tan ce i s t he re s pon s ibility of the s tud y Spo n so r. The fo ll owin g inf or ma t io n re lated to th e characteri za tion of the test s ub stance was prov id ed b y th e s pon s or. In format ion as pro vided b y Sponsor are as follow s : pH : 6.47 (at 1 % w/ v in aq u eo u s so lu tio n , ana lyse d in-hou se)
. Spec ifi c g rav it y / densi ty : Not App li cab le. Tes t s ub s t a n ce i s a so lid
.

EYE C opy No. 212 Pa ge 10/ 50

### 8.1.2 Test Sys tem A nim als So urce

No. of rabbit s Age at treatment Date of birth Bod y weig ht range at treatm e nt Id e ntifi cation

Rabbit access io n number s Accl imati za tion

8.2 Methods 8
. 2.1.1 Husbandry Co ndition s New Zea land Whit e Rabbit s Inh ouse Bred (Outbred) Toxico logy, D e partment of Safety Assessme nt Advinus Therapeutic s Limited Bangalore 560 058 , IND IA 3 male rabbit s 7
- 8 m o nth s 26 July 20 10 to 30 Jul y 20 10 2.61 to 2.94 kg By a nim a l access ion numb e r , cage label , c rys tal vio le t colour b o dy m a rkin g . During acc lim at iz ation the anima ls we re id e nti fi ed b y ear tattooing numb e r s . RB8767 to RB8769
On e rabbit (RB8767) was acclimati ze d for five da ys w hil e the other two (RB8768 a nd RB8769) rabbit s were acc lim at ize d for s ix da ys und e r la boratory co ndi t ions afte r ph ys ic a l e xa mination.

A nim als were h o u sed und er sta ndard lab o rato ry condition s , ai r cond iti oned w ith adeq u ate fre s h a ir s uppl y (12-15 air c h anges / hour). Enviro nm e nt: temp e ratu re 2 1- 23°C, relative humidit y 59
- 67%, w ith a 1 2 h ou r li g ht a n d 1 2 h o ur dark cyc le . The ma x imum and minimum temperature a nd relati ve
EYE C op y N o. 2/2 Page 11 /5 0

### humidity in the exper im e nta l room was recorded once dail y. The relative humidity in the exper im enta l room was ca lcu l ated from d r y and we t bu lb temperature recording s . HOll s ing T h e rabbit s were h oused indi v idu a ll y in rabbit cages (approx. s i ze: L 65 x B 65 x H 45 cm) w ith nory l s h a ll ow cage bod y and faci liti es for pelleted food (stain less s teel feed hopper) and drinking water (750 mL markrolon bottle fitted w it h s ip pe r tube). The li tter co ll ectio n tray (noryl waste tray) was c h anged dail y (except o n Sunday). The feed hopper s were c h anged once a week. Diet 

### ad libitum The a nim a ls were offered rabbit feed manufactured b y Pranav agro In dustr ies Ltd , Sang li Maha ras htra, In d ia. Th e d iet analysis report i s e ncl osed a s Annex ur e I
. Specific contami nant s (heav y metal s , orga n oc hl or in es a nd organophosp h ates) in the d iet are ana lyse d periodica ll y and the report are included in t h e s tudy file. Water 

### ad libitum Deep bore-we ll wate r passed through activated charcoa l fi lt er and exposed to UV rays in Aquaguard on -lin e water filter-cum-purifier , manufactured b y E ur eka Forbes Ltd., Mumba i- 40000 I , Ind ia was prov id ed to an im a ls in Markro lon bottle s w ith sta inl ess stee l s ippe r tubes. The water bott les were repleni s h ed o n ce dail y a nd th e water bottle s were changed once a week. The water anal ys i s repOit i s enclosed as A nn exure 2. 

### 8.2.2 Test S ub stance Instillation Prior to in sti ll atio n , both eyes of each rabbit were examined wit h the a id of a pen li g ht for irri tat io n and ocu lar defect s. T h e eyes were t h en checked for opacity using op hth a lmi c fluore sce in sod ium s trip s. The eyes we re t h en irri gated w ith e no ugh p hysio logica l sa li ne (0
. 9% NaC I) to remove any excess flu orescei n (approx im ate ly 30 seco nds). Us in g a n ul t ravio let lamp, the eyes were checked a nd eva luated for pre - exist in g corneal damage. No co rn ea l defects were observed in any ofthe animals. One hundred milligram s (O .l g) of the finel y g round test s u bsta n ce was placed in t h e everted l ower lid (conj un ctiva l sac) of the left eye of each rabbit.

EYE Co p y No
. 2/2 Page 12/50

### The eye lid s were gently held together for about one seco nd , in o rd e r to minimi ze lo ss of the tes t s ub s tance. The ri g ht eye of eac h rabb it remain ed untr ea ted and se rved as th e ref e ren ce control. 8
. 2.3 Anima l Restraint Eac h rabbit was re s train e d us ing an E li za b et ha n co ll a r fixed around the n ec k. One rabbit RB8767 was restrai ned for 72 h ours and the ot her two rabb it s (RB8768 and RB8769) we re re s trained for 48 hour s p os t-in s till ation. 8.2.4 S t epw ise exposure of A nimal s A si ngle rabbit was initiall y exposed to th e test s ub s tance. S in ce the te s t s ub s tance was not co rros ive, th e te st was comp leted us in g two add i t iona l an im a ls . 8.3 Observ a tions 8.3.1 Toxic Signs and Pre-Terminal Deaths The ra bbit s were obse r ved for toxic s ign s an d pre-terminal death s four ti mes (at hourly in terva ls) o n the day of te s t s ub s tance in s tillati o n and once d a il y till 72 hours po st insti ll ation. 8.3.2 Body Weights Body we ig ht s were record ed at the sta rt of acc lim at ization , Da y I of tre atment Gu st be for e test s ub sta nce in s till ation) a nd at s tud y t er minati o n. 8
. 3.3 Eye Scori n g Ocul a r les ion s we re eva lu a ted w ith th e illumination of a w hi te li g ht so urce at int ervals of approximately I , 24, 48 and 72 h ours p ost-in st illation of the te st s ub sta n ce a ccording to t he "Sca le for Scori ng Ocular Lesions" (D ra ize, 1 977)
. t he fluore sce in d ye eva lu ati on procedure de sc ribed in Sect ion 8.2 .2 was u se d to exa mine the treated eye of one rabbit (# 

### RB8767 , w hich sh owed cornea l op a cit y at 24 hour s) at the 24- and 48-hour obse rv a tion. At th e 72- hour observatio n, thi s rabbit had no opac ity a nd a ll it s eye sc ores were 

### O
. Hence , fluore sce in was not use d and the experime nt was terminated. T h e ot he r two ra bbit s had no cornea l opac ity, so their eyes we re not observed using op hth a lmi c flu oresce in
.

EYE C opy No. 212 Page 13 / 50

### Individual scores we re rec orded for each rabbit. A ll sco res were ca lculated a s de sc ribed in the "Scale for Scoring Ocular Le s ions", and the tota l po ss ibl e point s (out of 110) were tallied. [n addition to the observation ofthe cornea, iri s and conjunct i vae, any ot he r le s ions th a t were observed were noted. To aid in the interpretation of data , the average sco re for all rabbits at eac h sco ring p e riod was calculated (Appendix 2). 

### 9. RESULTS 9.1 Toxic signs and pre-terminal deaths Refer to Table I 

### There were no to x ic s i g n s or pre-terminal death s . 

### 9
. 2 Body weights Refer to Table I 

### The bod y weights of a ll rabbit s increa se d s lightly through the ob se rvation period. 

### 9.3 Eye 

### reactions Refer to Table 2 a nd 3 The s ummar y of corneal , iri S 

### and conjunctival reactions (Individual Eye irritation sc ore s ) and total s core s are pre se nted in Table 2 and mean eye irritation s core s are pre se nted in Table 3. 

### 10. CONCLUSION Under the conditions of thi s s tud y, caused conjunctiva l irritation and corneal opacity in the rabb it eye, wh ich cleared by 72 hour s. 

### 11. 

### ARCHIVING The original s igned final report w i II be se nt to the Sponsor and one s ign e d final report w ill be retained b y Adv inu s. The original raw data , documentat ion, record s, the origina l s tudy plan , and final report ge nerated a s a re s ult of thi s s tudy w ill be se nt to the Sponsor for retention following s igning of the final re port. Copies of the sa me (authorized by the Study Director) w ill b e retained by Advinus. Advinu s wi II maintain the se record s for a period of at least fi v e
EYE Copy No. 2 /2 Page 14/5 0

### yea r s . After this time, the Sponsor of the s tud y will be offered the opportunity to take po ssess ion of the record s or will be charged an archiving fee for continued archiving by Advinus or to dispose of the ra w d ata in consultation w ith the Sponsor. A representative sa mple of approximately one gram of the test s ub s tance ha s been se nt from the test s ub stance stores to the archives in the test facility before the first di spe n s in g of test s ub sta nce to stu die s. This sa mple s hall be sto red for a period of 2 yea r s from the date of thi s final report or till next GLP in spect ion, whichever is later, however not beyond 5 years. In the event of any inspection by regulatory authorities, seek ing original ra w data for verification, the spon sor would be responsible either to se nd the original raw data to Advinus Therapeutics Limit e d or to the regulatory agencies for verification. 

### 12. REPORT DISTRIBUTION The final report (original copies) w ill be distributed as follows: Spo n sor: One signed final report in original (Copy No. 2 /

2) 

### and one soft copy of the report in PDF format. Archives: One s igned final report in original (Copy No. 1 /

2).

EYE Copy No. 2/2 Page 15 /5 0EYE Copy No. 2/2 

### 13. TABLES

Page \6 / 50

### TABLE 1. Body Weight, Toxic S ign s and Pre-term inal deaths Rabbit

No.


### RB8767 RB8768 RB8769 M: Male

EYE Copy No. 212 

### Sex

M M M Body weig ht (kg) At Start of At At acclimatization treatment Terminat ion 

### 2.57 2.61 2.69 2.77 2.88 2.92 2.82 2.94 2
. 99 Toxic PreSigns terminal deaths None None None None None None

Page 17 /5 0TABLE2.IndividualEyeIrritationScoresRabbitElapsedCONJUNCTIVAIRISCORNEANo.TimeRednessChemosisDischargeTissue scorePupilTissueOpacity AreaofTissuescore(hours)(0-3)(0-4)(0-3)(0-20)(0-2)score(0-4)opacity(0-80)ABC(A+B+C)x2D(0-10)E(1-4)(ExF)x 5Dx5FRB8767II0 26 00 0NA0242I21000Ia420482II80 0Ia2107200 000 0 0NA0RB8768II026 0 0 0NA024I0I40 0 0NA0480000 00 0NA0720 0 00 000NA0RB8769I I026 0 00NA024I0I4 000NA04800 000 0 0NA07200000 0 0NA0NA:Notapplicable;a=ophthalmicfluoresceinsodiumstripsusedtoaidinevaluatingtheextentofcornealdamage.EYECopy No.2/2Totalscore(0-110)63018064006400Page18/50

### TABLE 3. Mea n 

### Eye Irritation 

### Scores Observat i ons at po st in st ill at i on
1 hour 24 h ours 48 h ours 72 h ours

EYE Copy No

. 2/ 2 

### Con junct iva Iri s

6 .00 0 6.00 0 2.67 0

0 0 Mean sco re Co rn ea Tota l mean score
0 6.00 6.67 12.67 

### 3.33 6.00

0 0

Pa ge \9 / 50EYE Copy No. 212 

### 14. APPENDICES

Page 20/ 50

### APP EN DI X 

### 1. C e r tificate of A nal ys i s

EY E Co p y No. 212**P~3ofl!l** 20fiS
CE RTIfICATE OF ANALYSIS FOR TE STIRE FERENCElCQNTR OL SUBSTANCES TI T L EIOBJECn\'E: QlMlICICt iutiOn**or** for U6(; IU' T CIl! and Reference Subs/mot TESTIRE FERENC£lCQ , "' TRO L S UBST ANCt;: DESCRIPTI ON:

SfRUCTlJRE: MOLECULAR WE IGIIT SnroV T N ITf AT I ONOA T £: ISJun e20 10 MET II DDS USEO: PURJlY : Gu**d1TO~IOgnpby** wi lli flame ioniution dt:Ieaioo (GC·FID) and Kul Fi$(:beT tilr.l.liQCl , IDENTIFICATION: Fourio:r TTJnsform- lntnred (FT · IR) specll'O$(:Opy &lid gas tlu-omaloi;f8phy-mau specuomcvy (GC-MS) RES ULTS AND CONC LU S I ONS; SIf'u(:(uraJ contirmaLion o f l>lc l<'II t**~Irnal** was pcrfonncd by FT·1R and GC·MS. PU RI TY DETERMJN All0N Rl'Sult ; alltreorrccting (or !he Kul**,** W1lcr result<
. onmR (Specify): 6$$ ppm wate r by Kar ] Fischer tirra tio n. No impurities were ckfttted at;:: 0.1% by GC-fTD anal)'$il
. CAL CU LA nONS: Arc_ Percen t ; '" (GC · FTD) Interm! Standw ; NlA fu:temal Sbnthtd: NlA Other (uplain ): Karl F ischer titr. u ion All origin.! IlIW dna associated with thilltudy were arc hived in the Sporuor rac il ity a«:h iv t
. Only descriptive ltati$tics wert: Died unle.n othe rwise no ted in th e mul lS
. Thif flooy was cond l.lcletl i.n lCCordmcc with the Good Labor.uory Practice StandMd. 40 ern. Pan 792 unteu**otherwi ~**  n ot ed. Due (0 LIlc short \(Tm natule orlbe study,**th~** s(Rbility orlbe t<:JI Sl.IbsLDnc:c 1Uldcl llo rg gc con ditions per 40 CFN. Pcut 792
. IO$(c) (U**l~e** test lit e w u nol verified.

Comple-tion Dalt : 

### 7(.,., / ;w r(l 

### ' Dati:



### __ 11. 

### <kL**""~**

### __ _

## o.<,rS T UDY D I R£CTOR AN D TEST INC FAC ILI TY ADDRESS: I mpact Analyti cal, 1 910 W
. St Andrews Rd
. Midland, M14 R640 T HIS IS A TRUE CO py OF THE OR IGIN/\, __**D.; ~A**  

### CAe. 12 - 2.3 - I D



### Pa ge 21 /50

### APPENDIX 2. N umeric al Scor in g Sys t em (Grades of Ocular l esio n s) (DRAIZE,1977) 

### 1. 

### CONJUNCTIVAE SCORES 

### (A) Red n ess (refer to palpebral a nd bu lbar co njun ct ivae excl udin g co rn ea a nd iri s) Ve sse l s normal
..................................................................... 0 Vessels definitely inj ected above normal
........................ .. .. .. More diffuse, deeper crimso n red, in dividual vessels not easi l y discernible
.. .. .............................. .... .. .. ................... 2 Diffu se beefy red
................................ .. ...... .. ................ .. ....... 3 (B) Chemosis Norma l
. .. . ........... .. ... .. ...... . .. . .. ..... .. .. ... . ... .. ..... .. ........................ 0 A n y swe llin g above norma l (includ in g nictitating membrane)
............................................................ .. ... . ........... I Obvious swe llin g w ith partial eversion of l id s .................... . . 2 Swe llin g wit h lid s abo ut h a lf c losed
.. .................................... 3 Swe llin g w ith li ds h a l f c losed to comp lete l y clo se d
.......... .. . 4 

### (C) Discharge No di sc h a rge .................................................................... . .. ... 0 A ny amount different from normal (does not in c lud e s mall amo un ts observed in inn e r ca nt hu s of n orma l an im als)
.. . .... .... ...... .. ... . .. .. .. . ... . .. .. ........ .. ... . . . .......................... . Di sc harge w ith moi s tening of the lid s and h airsjust adjacent to lid s .......................................................... .... .. .. ..... 2 Di scharge wit h moi ste nin g of the lid s a nd hair s a cons iderab l e area aro und the eye .... .. .. ................................... 3 TOTAL SCO R E = (A 

### + B 

### + C) 

### x 2 MAXIMUM TOTAL=20


### 2. IRIS 

## (D) No rmal
........ .. . .. ....... . ..... ........ ............... . ................................... . Fo ld s above norm a l , co ngest io n , swe llin g, C ircumcornea l injection (anyone or all of t h ese or Comb in at i on of any thereof) iris s till reacting to li g ht ( s lu ggish reaction i s po s iti ve)
.......... .. .................................... .. . No reactio n to light , haemorrhage, gross destruct i on (a nyone o r all of the se)
.......... .. .. .. .............. .. .......... .. .............. .. SCO RES 

# o

2 TOTALSCORE = Dx5 MAXIMUM TOTAL = 10

EYE Co p y No. 2/ 2 

### I 'age 22 / 50

### APPENDIX 2 eontd. Nume rical Scoring System (Grades of Ocular lesions) 3. 

### CORNEA (E) 

### Degree of 

### Opacity (most dense area u sed) No opac ity
................................................. . ........................... . Scatte red or diffu se areas- detail s of iri s clearly v is ibl e .................................................................................... . Easi l y discernib le tran s lucent areas , details of iri s Slight l y obscured
.......................... . .... . ....... . ........................ . .. . Opalescent areas, no detai l s of iri s visib l e, s ize of pupil barely di scern ible
................................... . .............. . ... . .. . Opaque, iri s not discernable through the opacity
................... . (F) 

### Area of Cornea Involved One quarter (or le ss) but n ot zero ............................... . .. . ...... .. Greater than one quarter but le ss than half
.............. . .... ......... . Greater than half but less than three quarters
........... .. .......... .. Greater than thr ee quarters, up to whole area ........................ . SCORES 

# o

2 3 

### 4 SCORES



### I 2 

### 3 4 THE TOTAL SCORE 

### = (E x F) x 5 MAXIMUM TOTAL 

### = 80

EYE 

### Copy No. 212 MAXIMUM TOTAL SCORE POSSIBLE 

### = 110

Page 23/50

### APPENDIX 3 . Study Plan

STCDYPLAK STlJ""DY PLAl, No . : SPTEST ITEM CODE BY TEST FAC I LITY : 0 08-26 STUDY COD E E"\'E
ACulE EYE**IRRlTATIO ~**  STL"DY**I~RABBITS**

S P - L'\c Cop:" :'\0. 1/1

EYE Copy No. 212 TEST ITE::>I:
STUDY K o.:

SPOl'SOR D ow Europe GmbH B a chtobel strasc;e 3 CH- 8810 Horgen Sw itzet - land

TEST FACILITY DE PARTME NT OF SAFETY ASSESSMENT ADVINUS TH ERAPEUTICS LIMITED POST BOX N o. 5813. PLOT No s . 2 1 & 22 PEENY A II PHASE. BANGALORE
- 560058 D-UIA

PagE" **1119**

Page 24/50

### APP EN DIX 3 contd. S t u dy P lan
C O;\T E;\T S
1. STUDY D E TAILS
......................................... . ...... . ........... 4 1. QUAL I TY ASSURANCE
...................... . ..................................... 5 3. G LP COMPLiAl'\CE
................................................................... 5 4. T EST PROC ED URE G UIDE UNES
............................................ 6 5. 

### AMENDMTh"T PROC ED URES
. . ...... . . . .... .. . . ............ . .. . ....... 6 6. S AFETY PRECAUTI ONS
............................. . . ..................... 6 7. O BJE CTIVE
................................................................................ 7 8. JUSTITiCA TION OF SELECTI ON OF TEST SySTEM
............... 7 9. JUS Tm CA TI ON F OR RO UTE OF ADMINIS TRi\' TION
............ 7 1 0. MAT ERIALS Al'\;n METHOD S
...................... . .......................... 7 1 0. 1 Mate ri al s ....................................... . ................................................ 7 10. 1.1 10.1.2 1 0.2 1 0.2. 1 1 0.2.2 1 0.2.3 10.2.4 10.3 1 0.3. 1 1 0.3.2 1 0.3
. 3 1 1. 1 \.1 12. 1 3. T es1 '!.)"')tC' Ill ... . .......................................................................... 7 Tes t !)lJb5ta nc e. informat io n .. '
........... , .............................................. 8 Meth ods
.............. . ........ . .. . .. . ........................................................... 

### 9



### ... . ........ . ..... . ........................................................... 9 T est**!)ub~ta ce** **.** i nst ill at ion .. , ........ "
......... , ... . ............ . .... "
.............. , 10 

### A.n.i n 1a i r es1raint
.................. . ............. . ........................................... 1 0 Stepwise e x pos\U'e of 311 illl.c.1 1 s ....................................................... 10 Observatio n s ................................................... , ........................... , 11 

### ( 'li. lliea !**o~erva ti** **ll S**  .. ., ........... , ......... ., ....... .. . ., ............................. 1 1 B o dy w eigh ts
...... . ........ . .. . . . ... . .. . .. . . . ..................... . ..... . ............... 11 Eye scor in g . .. . . ..... . ...... .. . . . .................... . ..... . ........................... 1 1 RESULT S
.................................................................................... 1 3 D ata cO Il tp il at i oIl ......... ............................. , .................................. 13 REP ORTIl\G
........ . .... . .... . . . . ..... . ................................................... 1 4 ARC HIVING
................. .. . . ........ . ............................................... 15 SP- E YE Co py;\o. l f l P age **2 /** **1 9** 

EYE 

### C o p y No. 2 / 2 

### Page 25/ 50

### APPENDIX 3 contd . S tudy Plan

14 . S TUDY PLAl'l DI STRIBUTION
................................... . ............. 15 1 5. REPORT DI S TRlBUTIO N
....................................................... 1 6 1 6. REFERENCES
......................................................................... 1 6 1 7. AGREE1viENT
............................................................................ 17 APPENDIX I. PROCEDURES FOR USE OF ANESTHETICS D URING

SP-G 7 69 ._EYE Copy**o. I**  **I I** 

EYE Co py No. 212 OCULAR I RRITATION TEST IN RABBITS
. . ... ................. 18



### Pa ge 26 / 50

### APPENDIX 3 contd. S tudy Plan

L STl:DY DETAILS Study Title**Te~t** s ub s tnnce Study Kumber Test Facili ty

Spon s or

Moni t oring Scientist

Study Pcnolluel S t u dy Director Tecluuca! Co
- ordin
... 'ltor Clinical Veter in arian Study Schedule Study initiati on date Experi.menta l staIting date

SP
- E YE Co p y 

### ="0. 111

EYE Copy N o . 212 Acute Eye Il1"itatioll Study in**~,bbit .**  ( 07694 Department of Safety Ass¢s,smeut Adv i ..llus Therapeutic s Limited Post Box No. 5813 , Plot No s . 2 1 & 22 Peonya II Pha s e , Bangalore 560 058 IN"DIA Dow Etllupe GmbH Bachtobelstra ss e 3 CH
- 88 1 0 Borgen Switzerland DOIUla Ande r s on TIle Do\v Chem..ical Company Toxicology & Envi.roJUuenta l Resetlrch & COlm.lt ing (TERC) Bui ld ing 5 727 N onlstowll Road. P.O. Box. 904 Sp r in g Hou s e. PA 19477-0904 Em.ail : DOIUlaA.1lderson ( @dow.COlll
P
. J
. Praka s h, M.Sc. S. B. Mohan Kutu<"1l", M
. Sc. S. Rmnesh
. B.v.Sc. & A.H.
07 March 20 II 09 March 20 11

Page -' /1 9



### Page 27/50

### APPENDIX 3 contd. S tudy Plan

Ace 1ll1l<1tizat ion
TrcatJllcnt Ob s e " rvatiollc,: Experimen ta l completion date Draft RepOlt Study completi o n d'\te Stmt : 09 Ma r ch 20 II E nd : 1 3 March 201 1 ( s ing l e a num l ) and 1 4 Mar ch 20 11 (a ddi t io nal two anulk,ll s)' Start: 1 4 March 20 11 ( single anin k,l) IS March 2011 (addit i on,,1 allum lis), Start: 14 Mar ch 2 011 L.,te,t by IS April 2011 Latest by 30 Apri l 2011 L.,t"t by 31 May 20 11


### * : \Vi1I be in cluded 

### if the te st**lIbstallce** proves to 

### be llo11 -c o n -o s ive and **I** or not a s evere in " itant depending on th e s ingle <1nim. al test. ob s ervation '). The cO llfinnatory te st I1l..1Y 

### be conducted in a sequential m4UUl e r using 011e an ima l per s.tep or u s in g two 3.1lunal s. The detail s of c o nfirm<.1 1 0 1

Y te s t wi ll be 

### included in 

### t h e n: po 1 1. 2. Q UALITY ASS li Ro\."\ TE TIle Qlk11 i ty ASSlm Ul ce Un it of Advinn s Th erape u t ic s Litllired , Bangalore
- 560 058. Ind ia
. will UlSp<ct th e s tudy. t he raw data. the draft and final re po lt. FUldulgs of all ulSpections will be rep orted to the Management and to the Study Director. lt lSpectio n date s and rep o tti..ng date s 

### will be entered in the study repot1 . The Quality ASSlu'ancc Unit h.a s re 'v ie wed th e final study pla n a ud 

### will receive a copy there of 3. GLP C O:lIPLJ.o\. "\'CE TIle s tudy will be perfo ll lled in accordmlce with the following GLP regulatiom; : • OECD Principle. ofGLP (as revised ulI997 ). ENVI MC/ CHEM (98)17. OE C D
. Park 1 998. • JMAFF GLP Standa rd s for Agricultmal Chemi c a k ll-Nou 5. 'n
No. 6283. I October , 1 999. • EC Dire ctive 2004 / I OIEC , Offi cial Jo u11lal of the European Union, L50 ! 44, Feb
. 20. 2004. • Thi s st ud y w ill a lso be cond uct ed as per the lllUtlk 'l ll y agreed s tudy plan and the Stand..'l rd Operat in g ProcedlU ' es,

SP
- EYI Co p y:"o. 111 Pnge 5/ 1 9

EYE Copy No. 2/ 2 Page 28 / 50

### APPENDIX 3 eontd. Study Plan Exception s to the GLP compliance are li s ted below:

• Archiving ofraw data at the te s t 

### £1cility . • Exception to guideline 405 , for not te s ting weight of evidence analy s i s by the te s t facility. 

### 4. TEST PROCEDUU: GUDELL,,\ES TIle prOCedlU " eS as described in thi s study plan are ba s ed on the following te s ting guideline s : • OECD Guideline for T e s fu12 of Che. lnicals, Section 4 , Number 405. ''Acute Eye ll.1'itation / (-011'0 5ion" adopted 24th April 2002. • U.S. EPA Health Effects Tes t Guideline s, OPPTS 870.2400 , Acute Eye**11l'itatio~** Augu s t 1998. • JMAFF 12-Nou s an-8147. N ovember 2000. • Officinl IOlunal of the European COll11Uluuties , Methods for the Detennination of Toxicity. Pmt B.5 (Eye In'itation), Directive 200 4! 73 IE C , 29 Apl' i! 2004. 

### 5. A.'IEl\lHIE:\T PROCEDUU:S All 3mclldulent s to and **I** 01' deviation s fiolu thi s study pIau and the rea s ons therefore shall be documented , d.1ted, s igned by ihe Study Director, approved by 1ll..:1nagelnent**a~** may be required) and n1.1intai..ned \vith the study plan . 

### If immediate action i s necessary , verbal agreeluent \vit.h the spon s or will be cOll.fll"llled as soon as pos s ible 

### by s tudy plan amendment. , AllY aluendlnem s to and/or deviation s 

### froln this study plan 

### will be recorded 

### ill the raw data and doclunemed in the [mal repolt. 

### 6. SAFETY**PRECAT:TIO~S** Glove s , cap , £,ce n1.1 s k and goggles (if required) will be used in addition to protective body gal'lucnts and shoes to ensure adequate per s on".,1 health and safety und to avo id uilialation and SkUl contact \vith the te s t sub!!.tance . 

### In ca s e of eye contact , the eye 

### will be wa s hed thoroughly with \vater and medica l treahue11t 

### will be s ought. In ca.':.e of skin**contact~** it will be washed with soap and \ : vater with ':.1.1b s equellt Iuedieal aid .

SP- EYE**Cop~'** :"0. III

EYE Copy No. 212 Pngt' (j ( 19

Page 29/50

### APPENDIX 3 eontd. Stu dy Plan 7
. OBJECTIYE TIle objective of this acute eye initation <o.mdy in rabbits i s to provide i..llfoI11k1tion on the potential for ul ' itatioll 

### arising fi :om a s ingle ins t illation ofa te st sub stance into the eye. 

### S. JUSTITICATIO:\ OF SELECTIO:\ OF TEST SYSTEi\I The rabbit i s the 

### syslem of choice becau s e. historically : 

### it ha s been the prefclTed 

### and most commonly used species for primar y eye in 'itatian tests. 

### 9. JUSTlFICATIO:\ FOR RO UTE OF AD:'IIL"\l:STR4.TIO:\ TIle ocular route of admin.istration 

### will be u se d becau s e 

### it is recollunended ill the referenced. guidelines and because hUllL'1n expostll'C 11L.1Y occur via this route. 

### 10. :'IrA TERIALS A,\'D METHODS 10.1 "fat. rial ,
Animal Source

No. ofrabbits Body weight rangeat treatment Age at treatment
I dentification

SP- EYE**Copr~o.l l** 

EYE Copy No
. 2 /2 New Zealand White R.1bbit s Inhou s e Bred (Outbred) New Z<aland White R.1bbits Toxicology Department ofSafdY Assessment Advinus Thenlpeutics Li.mited Bangalore 560 058 , INDIA 

### Up to 

### 3 Male rabbits 

### 2.2 to 

### 3.2 kg YOtlllg: adult (the exact age will be ment ioned in the report ) 

### By rabbit acces s i on mUllber. Identification of individt!.11 anink1 1 is by cage label,**cry~lal** violet I tunuel-ic colour body marking. Page 7 / 19

Page 30 / 50

### APPENDIX 3 contd . St udy Plan

Rabbit acces. s ion number s: Ac climatiz ation 

### RB8767 to 

### RB8769 After phy s i ca l examination for good health a.nd s uitability for experiment. the rabbit s wil l be ac ciuIJ.,tized for at least five days before treatment. 10.1.2 TE'st substance informatioll 

### (as fimu s hed by the Sponso r) Test s ubstance ! Item Lot 1'0. 

### Manufactured by (Nallle 

### and Addre<s) Supplied by (Name and Addre ss) Date of ll'laIlU£1ctw'e 

### Date ofexpuy PlU'ity a s per Cet1i.ficate of Analysi s Physical appearance Storage condition s 

### ( 201000599
- 6 The Dow Chemical Company Midland. MI 48674 USA
The Dow Chemical Company Midland. MI 48674 USA
March 2 010
06 March 2012 

### 99 .8 5 %

Off-white s olid

: Ambient (+ 18 to +36 ° C) The te s t ; ub s tance. identified as was received on 03 Febnk11 Y 20 11 and wa s fiuiher identified a s 008-2 6 b y te s t facil ity
. Refel'ence N umber dated 07 Febnk'lly 20 11
. The te s t s ub s tance is a s olid and i s sto red at ambi ent condition (+ 18 to +3 6 ° C)
. DOClUllentation of the l11eth ods of s ynthesi s . £1brication. or derivation 

### ofthe te s t s ubstance i s retauled by the Spons or. SP- E'l.L Copy;\o. 111 P.,ge- 8 / 19



### EYE Copy No. 2/ 2 

### I 'age 3 1/5 0

### APPENDIX 3 contd. Study Plan

Detenninatioll of the sta bility. identi ty . st rcngth ) purity (lud co m po')itio ll of the te s t s ub sta nce i s the re s pons ibility of th e st udy Sponsor. The follo\ving: illt ol1 u atio n related to the c h aracterizat ion of th e test subs tance was pr ovided 

### by the spo n so r . Informatio n as provided 

### by Spo n sor are a s follow s: pH : 6.47 (Ana l ysed in
- hou s e). Specific grav ity I den s i ty: Not Applicable. T es t
. mb>tance is a s olid. 10
. 2 Method s 10
. 2
. 1**H\lSba (h'~'**  Conditio n ') Anim"''lis wi ll be hou s ed lUlder s tallck 1 rd l ab orato lY conditions , air condit ioned w it h adequat.e fi' es h a i l' suppl y ( 1 2-15 ail' cl k1nges / houl'). EnvirOllluent: temperattu 'e 1 7 -2 Y' C 1 relative InUllidit y 30 - 70**%~** w ith a 12 hour light and 12 hom' d.. 1rk cycle. TIle m3 x llllulll and lu.iniuIll1U temperature and rel ative hmuidity in tlle eXpe!uuelltal 1'00111 will be recorded once daily . TIle relative. hmnidity in the experimental room will 

### be calcu lated :6: 0111 dIY and wet bulb temperatw:e recording s, H o u s in g Hou se d ind ividuall y in rabbit c ages (approx. si ze: L 65 x B 65 

### x H 45 cm) w it h n 0 1 yl s h.1llow cage body and £,cilities for pell e ted food (Stainless st eel feed hopper) and drinking wate r (750 ml mal'holon bottle fitted wit h s ippet· t ube)
. The litter co lle ct ion tray (notyl waste tray) wil l be changed cL1 ily ( exce p t on S 'Ulda y s). Diet: 

### ad libitum Rabbit feed m .a llu fac.hu 'ed 

### by Pranav a g ro Indu st ries Ltd , Sang li , Malk'tra s htra
.. India , w ill be provided 

### ad libit llllJ to the rabbi". The proclu'ed feed wi ll be s ubjected to analysi s of cOluposition and the analys is rep0l1 incl u ded in the repot1. Spec ific co ntaminan ts (heavy 111 eta l s organochlorine s an d organopho s plk1tes) in the di et al'e analy sed p er io dically and the repo 1 t 

### w ill be included ill the study fi l e .

S P -G'6 94

_ EYE**Co p~** **·** Xo. III



### EYE Copy No. 2 / 2 Page **9 /** **19**

Page 32/ 50

### APPENDIX 3 eontd. Study Plan
Water: 

### ad libitum Deep b o re-well water p asse d through activated charcoal filter and expo s ed to UV rays in Aquaguard in-line water filter-cum-plU ' ifier manufactured by Elu'eka Forbes Ltd
.. Mlunbai
- 400 001. INDIA , will 

### be provided
. TIle water bottle will be repleni s hed once daily and the water bottle s changed once a week. The water alwly , is re snlt s will be fiunished in the repol1. 10.2.2 Test substance in st illation Prior to instillation , both eyes of each rabbit will be examined with the aid of a pen li ght for in'itation and ocular defects. TIle eyes will then 

### be checked for opacity u s ing ophtlk11mic fluore s cein sodium test s trips. The eye s will then be irrigated v .. ith enough phys iological sa l ine (0.9% NaCl) to remove any excess fluorescein (approxink1tely 30 seconds)
. Using an ultraviolet lalnp. the eye s will be checked alld evaluated for pre-existing comeal ci1nk1ge. Rabbit s show ing any defect s will be eliminated and replaced. For solid test snbstance: 0.1 g of the finely grolUld te s t item wil l be placed in the everted lower lid ( conjlUlctival sac) of the left eye ofeach rabbit. TIle lid s will be gently held together for about Olle s econd. in order to minimize lo ss ofthe te st s ub s tallce. If it i s knowll tbt the test s ub stance may cau s e comeal opacity or pain il1'itatiolL or if pain or di s comfo11 is noted after any anuml i s in stilled , the procedure s for lese of anaesthetic will be followed as de s cribed Ul Appendix 1. 

### A.'1L'1IAL 

### SELECTION Stepwise 

### st arting 

### with a single anima l: Selected 10.2.3 Animall'esfminf TIle Elizabethall collar will be fixed around the neck of rabbits and retauled until the eye s core, 

### if any 1'enU11 to zero 01' till 

### 72 holU's, 

### whichever is earlier. 10.2.4 Stel",ise exposure of 

### animals A 

### single rabbit will initially be exposed to the te st substance. Prior to in s tillation of the fe st s ub s tance , a local anae sthetic nwy be used in both t he test and control eye. 

### If a cOll"O s ive effect is ob served at S P
- EYE Copy**~o.** II I



### EYE Copy No. 212 PagE' 10 / 19



### Page 33/50

### APPENDIX 3 contd. S tud y Plan
fUlY sco rin g interval. the te .<.t will be tenninated llllluediatdy and no fUl1her testu l£! will be conducted
. If no conusive effect i s observed <lnd inltation d oe'S not**s~ t**  tluollgh Day 21. the te s t will be c o mp l eted u s ing two additional an.im<1 I s. 10
..3 OIJ SE' lTatioll S 10
. 3 .1 Clinica l obs("l'Yatioll s Rnbbits will be ob se rved fOl ' toxi c sign s and pre-te
. muual d eath s fotu ' times (at hotll'ly inrervab, ) 011 the d.1.y of te st s ub sta nce im.tillat ion and once daily thereaftet' till the end of observation. Ob serv ations will include gro .'.s evah l a li on of s kin and fi.u ·) eyes and mucous. membrane c... re.

spiratolY. ci.rcu1.1tory , autonom.ic and central nervollS sys tenls. 5.0 matOlllotor act ivity aud behavior pattent
. P a lti c ular attention will be directed to ob s ervation of tremors
. convul s ions ; sa livation
. di n n 'h ea and COllla
. 10.3.2 Body weight s TIle body weights will b e recorded at the 5ota11 ofaccluuatization. d'\y 1 of t reatment Gu st before te s t s ub s tance ul50til l ation) and at te.J"Illiua tion of observation / death
. 10.3.3 Eye s(' ol'ing OcuL."1f**le oll~**  will be evaluated wit h the illumiuntion of a white light sO \U'ce at interva l5 of approxim
... '\tely 1. 24 , 48 and 72 hom's po st instil l at ioLl of the test s ubs tan ce accor ding to the "Sca l e for Scoring Ocub,. Le SiOll S' · (DRAIZE
. 1 977). 1. co',.n;:-;CTIYAE SCO RE S (A) Redue 'is ( refe-r t o p a lp eb ral .,nd bulbar co u j utl c lI\'ae)
. Ve-s s el s JlofllL'"\l
.. . ... . ..... ......... ... . .. . . . ..... .................. 0 Vesse ls definitel y in j ec t ed abO\'e n orm,,'!
... . More diffu 'ie , deeper c riJu s: ou red. individual vess:e l s not ea s ily di'>cernible
. ................ . . .. .. ... . . ..... .. . 2 Diffu se beefY r ed
... ............ 3

S P -Gi6 94

_EYE**op~ '**  :'\0

. II I

EYE Co p y No. 2 / 2 

### Page 34/50

### APPENDIX 3 contd. S tudy Plan

(B) C ht>In osis No rnm}
................ . Any !;wd hn g above n OIul<1.1 ( in cluding nic titating: tue1ll.bnule)
. 

## o
Obvioll S s'welliug \
... illl pat' tial e. , ' er s ion of lids ... . .. 2 Swell in g with lid s abom h a l f closed
.. . ........ 3 Swell in g with lids half closed to completely clo sed 4 (C) Di 'i l" har ge. No d:i-.:charge
. . .. .... ............................................. 0 Any am ouut**diff~ent** from norm.al (does not includ e 5111<\ 11 amolln t s obseryed In ilU l er canthu. s of ll O fllli'll alum
.. ,ls)
. . ............................................... . Di ':>c harge- w ith moi '> ' t!' lU l1g of the- lids and h air<; j U 5t ndjn ce nr t o lid,
.................. .. .. . .. . ....... ... .. 2 Di sc harge with Ulo i'> tenin g of t h e li ds and h <1.lc.c:.: a Cons i derable a r ea around th e eye ....... .............. . . ......... ....... 3 TOTAL SCORE**~( B**  +C) ' 2**~L·\XL\IV?I** TOT AL**~** 20
2. IRI S (D) Fold s above n orma l. congestion. swelli n g , Circulllconlc-al**i.llj~tion** (a ny oue or all of th e<'j e or C'ombiu.1tiotl of all Y ther eot) iris still reac t ing t o light (sluggis h reacti o n i s positi"'e)
................................. . No r eaction to light
. ha emorr ha ge , g r os s de s ttu ct ion (a n yo n e- or all of these)
.............. . SC ORE S 

## o

2 TOTAL SC ORE**~** D , 5 1\L-\....x: nIl 1\ I TOTAL = 10 3. CO R.'\ EA ( £.) D eg n..e- of**Op:lcit~ ·**  (most dense area u se d ) No opacity ScatTered or diffue;e areat; - detaile; of iri s clea rl y nsible Ea s ily di sceru ible tran s lu cent areas. detail s of iris SC ORE S 

## o
Clearly vi')ible
. ....... .......... ..... . ...... . ... ... ... ...... ..... 2 Opale')cem area'). no details of iri s , , ·isibl e. size of pup i l bardy di

.. cern ibl e .... . ............... 3 Opaque. iris mvi t; ibJe
... . .......... ...... 4**SP-G7 69~_E.Y.E** **Cop~'** ::'\0. 1/1 Page- 1 2i 1 9



### EYE 

### Copy No. 212 

### Page 35/ 50

### APPENDIX 3 contd. Study Plan (F) AI'(>:1 of C:Ol'ut':l !U\ " oIYE'll One quartet" (or I e ':> !» but not Ze1"O
. . ... . ........ , _ ............ . ......... . Greater than one quartet" but le s s than half Greater th.1 n halfbut le <;5 than thre e quarte1" <; Greater tl1..1n thre e qu ar ter .... up to \"h o l e nr e a THE TOTAL SCORE = (E' F) x ;. 2 3 4**)L-LTI\Il~1** TOTAL = SO**)L-L'U)Il~1** TOTAL SC ORE PO SS IDLE = 110 

### In addition , all treated eye s wil l be 

### eX.1lnined u s ing ophthal.mic fluorescein so dil1ll1 at the 24-hotu' interval. If cOluea} opacity exi s ts at 24**h01U'& ~**  fluore sc ein \vill be ll s ed , as needed.. at s ub s equent scoring intervals to aid in 

### eVahk'l.ting: t.he extent of 

### c01l1ca l d-11uage . If there is n o evidence of ocu lar in 'itation in a ny te s t rabbit at 72 h om's. the study will 

### lx tenllinated. If irritation pert:.ists in any rabbit at 72 

### hom '" s coring will 

### cont inue fol' all rabbit s at 4, 7, 10, 14 , 17 

### and 21 

### days post-instillation or \uttil all signs of in'itation ru'e completely rever s ed (n sco re of zero ) 

### in all rabbit s . whichever OCC1U'S first. The dlll- arion of the sco rin g period will not exceed 21 

### days unle ss 

### authorized by the S POllSOL Individual scores will be recorded for each rabbit, All score; will be 

### calculated as de scribed in the "Scale for Scoring Oculru' Le s ions". ru1(\ the totnl pos s ible points (out of 110 ) 

### wi ll be tallied, In addition to the ob.,ervation ofthe cornea. iri s and conjunctivae , any other le s ions that 

### al'e observed will be noted , To aid in the inte1pretation of data , the average score for all rabbit s at each scoring period will be 

### calculated, If

, at any tune dtu-ing the st udy , extensive in- itation i s noted and 

### deemed in'ever s ible, the Study Director will notify the Sponsor thM the st udy shou ld 

### be tenuinated. If dallk'lge is exten s ive and the Sponsor 

### is lUlAvailable to authorize termination of the st udy. the Study Director 

### may unilaterally decide to have the animal> euthanized inullediately for hUll1.1Ue reaso us 

### and the Sponsor wi ll be notified as SOO l1 as 

### po",ible, 11. 

### RESL"LTS 11.1 Data 

### compilation The fo llowing ck,ta will be 

### recorded on ck,ta sheet s and tran s cribed for CO lllpilation and a naly s i s: de sc ription of the degree and nature of in ·itatioll. the pl'ec,ence of ocular lesions. and any effects other than 

### oculru' noted during clinic al obs ervations and body weights, All SP- EYE Copy**~o.** III

EYE 

### Copy No. 2/2 Pag(> **13 /** **19**



### Page 36 /50

### APPENDIX 3 contd. Study Plan fmding; will be recorded m the stall dru'd repolting fonuMs an d presenred in the repolt. 

### 12. 

### REPORTL,G The fmal repolt will include the following infonnation: }- The n ame: and addrc:o;. s of the spoll.'!.or and the te st facility [liong 

### with the s tudy schedule. :,... The llames and s ignattm!s of all per so lU1el i.nvolved 

### in the S.tudYl 

### including the Study Director ruld other s cienti sts. "; TIle test 5. ub')tance with identification either 

### by name and **I** or code.**~** 

### PlUlty
. composition and other appropr;'1te chru'acteristic s of the te s t s ubs.tance .

,. Te st <;.ubstance :

• Te s t s ub s tance 

### inf0I111ation as prov-ided 

### b y the Sponsor



### , Vehicle:

• Identification. concentration (where appropriate ), voltulle 

### u ') ed .**~** 

### Te st anima l s :
• Specie sls tmin use d • Age of allinlal., at strut of treanllent 

### • Ntl111ber of animals and 

### sex • Individual animal - weights at 

### s trut and cOllclus.ioll oftest • SOllrce. 

### housing: conditions , diet , e tc



### , Re s ult s:

• Description of method ll s ed to score ini lfl tion at each observation t ime ;,.. A**~UnUll.c11y** 

### and de sc ription 

### of all re sults. .

,. All s tudy 

### plan deviations. if 

### any.

,. Individual toxic s ign s and gro .s.s 

### necrop sy**fUlding~ ~**  

### if any . SP
- EYE Copr**~o.** III

G71i94-EYE 

### Copy No
. 2 / 2 Page **14 /** **1 9** 



### Page 37 /50

### APPENDIX 3 contd. Study Plan**~** A de s cription of all ci:rcmllstancec; 

### if any , that lllay have affected the quality or integt'ity ofrhe s tudy. 

## »- Quality A'i s urance Statement. s.igned 

### b y the Quality A'iSUrallCe U nit. 

# » GLP Compliance Statement. sig ned 

### by the Study Director .**~** 

### The sTOrage 

### location of all raw dat.a , the report. a sample of te s t s ub st ance and the archiving period. ;;.. GLP Cetiificates
.

TIle 

### original s igned final report 

### will be s ent to the Spons or and one sig ned fmal repo11 will be retained by Advinll s. The original raw data , 

### docwnentation, record s, the original study plan , and final repol1 generated rI'3 a 1'e 5 01t of thi s s tudy \vi ll be s ent to the Spon s or for 

### retention follo\ving s igning of the fUlal repOlt . 

### Copies of the sa Ine (a uthorized by the Study Director ) will be retained by Advmus. Aclvinn s will maintain**th~~~ r~cord** **s** for a period of at least five**y~ars _**  After tills time , the Sponsor of the s tudy wi ll be offered the opportlUllty to take po ssess ion 

### ofthe record s o r 

### will 

### be charged an archiving fee for continued ru- duving by Advi.lltl s or to di spose of the raw data in con'S:l1ltation \vith the Sponsor_ A repre se ntative s alllple of approxlluateiyone gt-anl of**th~** test itenl will 

### be**~nt** :£i:om the te st irenl s tores to the archive s in the fe st £1cility before the fu-st dispensing: of te st item to st udie s . Till s s ample sh all be stored for a period of 

### 2 years fi'om the date of rlll s fmal ropo1t or till next GLP inspectiolL wlllchever is later. however nol beyond 5 year s _ In the event of any**lllsp~ction** 

### by regulatOlY**auth.oriti~ !.;**  s eeking o ri ginal raw data for**verificati ~**  the spon so r \vould be**re spo** **n~ible** either to seud the origina l raw data to Advinu 5 Therapeutic s Li.mited or to the regulatOlY ag\'!"llcies for verification_ 

### 14. STL'DY PLA
.. " 

### DISTRIBrnOl\' TIle f11101 study plan (o r igina l copies) will be di st ributed as follows: Spon s or: Soft copy of t he s igtled Study piau in PDF format and one s igned s tudy plan ill original (Co py No. 1 1 1 ). Study Director: One s igtled study plan in original (Copy No. 

### 111 )
. SP- EYE Copy**o_** 11 1



### . EYE Copy No. 2/2 Pilge **15 /**  **19**



### Pa ge 38 /50

### APPENDIX 3 contd. St udy Plan 15. REPORT DIsTruBl
-no:" 

# n," fUl a l repOlt (original copi,, ) will be di s tributed a s follow;: Te s t Facility Archives: Olle 'i igned fUlal repo l1 in orig in al (Copy No. 

### 112). SpOJlsor: Oue s igned fm a l repolt in original (Co py Kos.2 / 2 ) and one Soft copy ofthe r<polt in PDF fonnat. 16. 

### REFEREl\CES 1. Draize. J.H
. , Woodward, G and Calvely. H.O 1944. J
. Phal111ac ol. Exp.TIler.83: 384. 2. Draize
. J.H 1 977. Dennal Toxicity In: Appraisal of the sa fety of chemicals in Foods. Dn. lg s and Co s metic s. As s ociation of Fo od and Dmg Officials ofthe United State s . Austin, Te x.1s. p . 47. 3. OECD. 2000. Guidance Document on the Recognition. A ss e ss ment and Us.e of Clinic.:'l l Signs as H lIluane Endpo int s, for Experimen ta l Anim•. 1.1s Used in Safety Evaluation Envu ' olllucntal He alth and Saf ety Monograph Series 011 Testing and As s e ss ment No 19. (ENV / JMiMONO (2000) 7.

SP- EYE Co p y:'\o. 11 1

EYE C opy No
. 212 Pag(' 161 1 9

P age 39 / 50

### A PP EN DI X 3 contd . S tud y Pl a n

1 7
. AGREEMEN T Thi s S tudy Plan for Sludy No .:07 694, " Be nzoylated Calecho l: Acu t c eye irritalion s tudy in R a

bbit s" has been mutua ll y agreed and sjgned. Sponso r S tud y pl a n A ppr ov al: 

### nlC (inal version 

### o r the s t udy p lan 

### wa::; approved 

### by cJnai l 

### frolll the study mo nitor on 07 March 2011 for Study Dire c to r's Signature.



### ror TEST FAC ILITY

STUDY DlRECTOR D a!e: 7'" ,,,,-,,., 2<> I,

QUA LI TY ASSURANCE UNIT Da {c:Qe , mnrnf1 ,**)~"**

MA T AGEMEN T Da te: ()**~tI\«d.V'** I

S P -G769 ", _EVE Copy No. I I I

EYE Cop y No. 212 I' Rg(' **1 7/**  **1 9** 

Pag e 40 / 50

### APPENDIX 3 contd. St udy Plan APPL\lHX 1. PROCEDl."RES FOR USE OF .<L'\ESTHETICS DVRl;,;G OCTLAR IRRITATIOX TEST Il'\ RABBITS**C\ot~** : The- pl ' o(eclul'£>s d e s cribed are gE'u(>ral iu U:ItUI'!;' : md only those pl"oC"E'rhu'e s applicable ba s ed au the u;'lture / C'h:u ' ;lcteristic s of the te s t s ub s tance will be followed ) Pre-Application If the pH or formulation of the te s t**~ub tallce**  is 

### s Ulular to other te st compound , (previou,ly te s ted) that h.we produced comeal opacity or pain uTitatiOlL 2 · 3 drops of Lidocaine 4 % will be ulS tilled Ulto both the treated and control eye prior to do s ing . Dow will provide the infol1uatioll 

### regarding the previoll s ly**h: ted**  material. Post - application Lidocaine 4 % will be. ini ti a ted inunediateiy after do s ing 

### if exce s ')ive pawlllg: at the eye OCCllI'S (in the event of po w der admini s tration. the ph ys ical propertie s of the powd e r may c au s e trans ient pawing , there**~** not a need to treat in this ca s e) or 

### if pain re spons e i s more s ignificant - exce ss ive vocalization or trying to e s cap e . Two to three drop s of Lidocaule 4 % will be ulStilied Ulto both the treated and c o ntrol eye. Que hour Examination No TreatnlS:ut : 

### If there 

### is no or s light conleat opacity ($ core s of 0 , 1) , if there i s slight conjunctival redne ss ( s core s of 1. 2) . slight**cOl~ullctival** s we llul g ( score s of L 2). discbarge ( s core s of L 2) and u · iti s (score s of 

### O

. 1 ). The rabbit s will not be treated with an . 1e sthetic or analge s ic. OR Initiation oftreatmellt with Lidocaine 4 % anae sthetic (evelY 2 holU'S): Treatment 

### with Lidocaine 

### 4 % 

### is initiated 

### if the 

### rabbit i O) ob s erved with 

### s ignificant cOllleal 

### opacity ( s core s of 2, 3. 

### 4)1 cOluunctival s welling (s core s of 3. 4 )
. di , charge (s core of 3) , conjunctiv a l r ednes s (s core of 3 ) and iritis ( s core of 2 )
. Two drop s of Lidocaule 4 % will be ul s tilled evelY two homs, tJu-ougbout the workdny . to both the t",ated and control eye. R.1bbit Re · e\ ·alllated at the Eud of**Da~ ·**  0 if Lidocaine oj % treatment has be-en initiate-d Tennin"tion o f Treatment : If there is no or s light come a l opacity (s core s of O
. 1)
. if there is s light conjunctival redness (s core s 1 , 2) , s light conjunctival s welling (s core s of 1 : 2) or di s charge in the ab s ence o f si gllificnnt fUldulg s ( s core , of 1 , 2 )
. SP· EYE Co p )' Xo . **I I** **I**

EYE Co py No. 212 P.,gE' 1 8.1 19

Page 41/50

### APPENDIX 

### 3 contd. Stud y Plan
OR Initia te s y')temic tre atment: If oc ul ar in1.ta tion ha s wOl'<:.ened o r remained the s ame fro m the One Hotu· Eva ltL:1 tio llo; admin.i s ter BuprellOIphine (0.0 1 to 0
. 0 3 mgikg b y IVIIMlSC) at the beginning an d elld ofthe wo rkd ay . R E'-f TaluatioJl of rabbit dUl'ing t h E' fir s t 7 2 h ours po st clo seIn

itiate s yst emic tre atme nt : Treatment \\o"i th Buprenorphiue at the beginning and end of the work day (approxi mat ely 1 0 -1 2 ho ur interval)
. is initi.,ted 

### if the rabbit i s observ ed with significant comeal op acity ( ;core s of 2
. 3. 4)
. conjunctival sw elling (score s of 3
. 4)
. coqjunctival r edne >s ( s core of 3 ) and iritis (score of 2)
. T wo to three drop s of Lidocaine 4 

### % nk1y al s o 

### be u s ed prior to staining of the eye \vith fluorescein , an d rin s ing. to alleviate any pote nt ial 

### discomf0l1 that tills process (spe c ifically 

### ti- on l the 111 echmucal a s pect of rin s ing. the e ye) nl<'y cau s e the rabbit. TI,e topica l anae st hetic w ill b e administered at le a s t 30 seconds prior to the u sc of th e flu o r e s cein . TE'l'lnillatioll of**S~·stemic** Tr e atment Treahllellt lllay be s topped w hen co n ju ncti val uritation**~** comparable to the olle- h otu ' evalua tion cr it eria for n ot. tre ating d Ie eye with Lidocaine 4 % . and n o conlea l defect s are ob s e rved ( greater tban a score of 1) after s taining w ith fluore s ceul. I f at any time. the aninlal appears to 

### be in u ndue pain/di stre ss ( as indicated by clinical o b s ervation s confU"llled by a St udy Director arullo r Study veterin a rian ) : the an;'1I<,1 will be euuUUlized and it will 

### be noted in the study record s.

SP
- G i 694

_ EYE Co py::'\ o . **I I**  **I**

EYE Copy No. 212 P nge **1 9**  **/ 1**  **9**

Page 42 / 50

### APPENDIX 4. Deviation from the Approved S tudy Pilln Study No. St ud y T itl e Acute Eye Irrit at i on Study in R abb it s Tes t Item Co d e D08-26 S I. AS IN STUDY PLAN DESCRIPTION Ol? DEVIATION No. I

. 

### Pages 

### 4/ 19 and 

### 5 / 19 1. STUDY DETAILS STUDY DETAILS St ud y Sc h ed ul e: Study Schedu le: Draft re p o rt: Late s t by 30 April 201 1 Draft report: 25 May 2 011 Study co mpleti o n date: Latest by St ud y co mpl etio n date Latest by**I** 

### May 20 1 1 30 Jun e 

### 20 II



### - -------,-- 

### Exp lanati on : 

### To in c lu de the cha n ge 111 

### d raft report s ubllli ss ion da te a nd st udy co m 

### le ti o n date
. ----------- 

### 2. Page 

### 13 / 19 10.3.3 Eye Scor in g Eye Scor in g In 

### addition , a ll tr eate d eyes w i ll be In addition, th e eye of o ne r abb it w hi ch exa m ined lI s in g 

### ophtha lmic fluore scein s h owed cornea l o pacit y at 24 ho ur s was so dium at th e 24- hou r int erva l. If co rn ea l exam 

### i n e d lI s in g o phth a lmi c 

### flu o re sce in opacity ex is t s at 24 h ou rs, fluore sce in wi ll sod ium at the 24 a nd 48 ho ur int e rval. At be u sed , as needed , at s ub se q ue nt sco rin g 72 hours obse rvatio n , th is rabbit had no in te rvals to a id in eva lu at in g the extent of opac it y a nd no eye sco res and he nce co rn ea l dam age. 

### flu o r escein was no t use d and te rmin a t e d th e ex p e rim e nt. Ot h e r two r abbi t s w he r e ther e was 110 

### cornea l invo lveme nt , - ve re 110t 

### obse rve d 

### u s in g opht h a lm ic nuore s cein. Exp lanation : 

### To co rre ct and includ e the rabb it eye exa minati o n u s in g opht halmi c 

### fl u o r

escei n.

Date: 

# Ib 

# 'It) "-' 2.0 t(

EYE Copy No_ 1 12 

### (P. 

### J. I)rakash) Stud y Directo r

Page 43 / 50

### EYE Copy No. 2 /2 

### 15. ANNEXURES

Page 44 /50

### ANNEXURE 

### 1. 

### Analysis Report
- Animal Diet Sample

A D VI NUS T IT E RA I' EU TI C S LIMIT E D 2 1 & 22 , P EEN YA I N D US TIUAL Alili A , IlI'II ASE I3 ANG ALOR E
. 5 60 058 ANAL Y S I S n E l'On T ( A M E I'O J<: O )· A N IMAL IlI E T S A M P LE
F ro m : A na ly t i c a l R & D D epartm e n t A d v i n u s, D a n w tl ore- 5 6 0 058 To: Departm e n t of Sa f e t y A ss e s s m e nt Ad v i nu s , B :ul ga l o r e- 5 60 05 8 Our R e f
. N q. : W C n T S 12 14 7 D ate : 1 4. 0 6
. 2 0 1 1 S amp le D eta i ls : Name : R a bb i t F ct'd S a mpl in g Date : 19
. 10
. 2 0 1 0
B a lc h N o . :**05 9~**  S u pp li e r : P ran nv A g ro Ind u s tri es Ltd
. , Blln g alor c .

, Man u fa c lur e r Pralluv A g ro I nd us tri e s LI ' d . • S ll l1 g l i
. i\1a hmash tra . I nd ia

S I. N o . Paramet e r A NALYSI S R ESU L T S (An:lly s i s o n " ; I S i !i ba s i s ") ( '/o j**---I~-- -M** **;;'~ ---** **-- --------------------------------** **----- iT** **-'--'i:-- -C** **~d ;;;-cN**  6
. 2 5 

## :;'--
------ -- --- ----- -------- --- ------------------20.32 ·· 3 , Cru d e f al**(E ;'** **xtr ac** **r--- --** **------ --'-** **-- ----'** **-- -"-'** **--" -'--'** **''' ''** **'-'- ---** **'j '~i** **-"" -'''-** **4~ ''''-** **C~ fi** **b;'~ --** **''- '-** **''''- --** **'' --'----''--'-''**  

### ··· ···· --···--·-----·· · ·

.. -·--..**·· --·--** **·- ~98**  

### .-

.**···-- ---** **··· To** **l~ j"I** **·-···-···-·** 

### .. ·-·-·-··-- ·---··- ·- ··--· · ·-··-·· ·· ·-··-- · ··-·-· · --· ..**··-· ··** **··- -·······--** **~3)-"** 6

. A c id i n so l u bl e fi s h 

### --- .... _ ..... -.-- -'-"

..... ' 43"2 -'

8
. Cn l c iulll (en ) 0. 5 4

Rt! aso n f o r A me ndm e nt : T o**co ~c** **l** t h e M a n uf a c ture r 's ad dre ss .



### ·EYE Copy No
. 2 / 2 Anal yt i c al R&D O c pat"tm e n t



### Pa g e 45 / 50

### ANNEXURE 2. Analysis Report
- Water Sample

A D V I NUS TH E H A P EU TIC S LIMITED 21 & 22, PE " ENYA [ NDlJSTRIAL AR EA, II P I
- l ASE BANGALORE 560 0 58 ANALYSIS H E PORT· WATER SA MPL E From: A na l yt ical R&D Department Adv in us, Banga lo re-560 058 To: Depm1menl of Safety Assessment. Advin u s, Bangalore-560 058 Our Ref. No: WCrrWS1257 D ale: 24/03/201 1 Sample Details: Source of Collection: Outlet of the Aquagua r d (At use point) Date of Collection: 1110212011 ANALYSIS RESULTS 5 1. Parl'lmctc
. " No.

I. Colo ur 2. Odour 3. Turbidity 4. PH 5. Electrical Co n ductivity, dSm- ' 6. Total solids , (ppm) 7. Sus p ended so l id s, (ppm) 8. Dissolved solids. (ppm) 9. Dissolved oxygen (ppm) 10. Biochemical Oxygen Demand 5 days a! 20°C. (ppm)
. II

. Chemical Oxygen Demand (PPn'I)

EYE Copy No. 212 COli t-e n I Colourless Odour J ess Clear 7.36 1. 324 963 18 946 4.9 2
. 1
6.2 SI. No. 1 2. 13. 14. 15. 16. 17. 18 19. 20. Parameter Con tcnt m Total hardness as 220 CaC0 3 Calcium a s**Ca- ~**  37 Magnesium as Mg
-- 31 Chlorides us CI" 283 Sulphates as**SO~** - 50 Carbonates as C0 3 -· Nil Bicarbona tes as HC0 3 · 29 0 Sodium as Nil. 24.8 P o ta ssiu m as K 2.79

Atwlytical R&D Depat1mcnt

Page 46 /5 0

### ANNEXURE 3. GLP Certificate
- Germany

I I' " I Bundesinslilut fOr Risikobewertung I Federallnstitutc for Ri sk Assessment 

# FlcB fR

GU TE LABOR P RAX I S 

## I GO OD L AB ORAT ORY PRA CT I CE (gernM I according to § 1Ob Alls

. 2 Nr .3 Chernikaliengesetz I German C hemi cals Act) bne 3 LP

. lnspektion wurd e durcl,geftihrl in , A GlP inspec fio,; w as ea rned out at P r ilfein r ic ht ung I Tes t Facility Adv inus Ther ilp" IhcS Limitert ( f o rm er n a m e un ti l Octo b e r 201 0; Advinu s Th e ra p e u tics P ri va t e Li mite d) 2 1 & 22 Ph aso II , P c- s ny a In d u s t r ial Are a B an ga l ore - 56 0 0 :'; 8 , IN DIA P r Ofkat e goricn I A r ea o f Expe rt ise PrO fungen zur Bes t immung dcr physikarisch -chemischen Eigenscl1aften und Ge h aUs b es timmungen I P h ysical -dlemicallest i ng Pliifunge n zur Bestimmung der toxikologischen Eigenschaflen I Toxicity studies PrUfung e n zur Bes t immu n g**erbgutver~ndernden** Eigenschaften (if) vilm, in vivO) I Mu tagenici ty studies OkCIOxikologische PrQfungen lur Bestimmung dar Auswil k ' !(lgen au f aqua l ische und lerrestrische Organismen I Environmental toxicity stud i es on aquatic and terre strialo rganismr: Pruflln!)en Zlim Verhallen im Ooden

. im WSSE o r lind in dcr Luft : I3lookkumulolion I**Slud o.:~**  0 11 behavio u r tn soit, water and air: bioaccum u lallon PrOfung en zur Bestimmung von RO(;kstanden I Residue studies Anal ytische PrU fungen a n b iologisc h en M at e r i a lien I Ana l yllcal and clinical chemistry testing Sonstige PrOfungen
- Blonanalytik Pharmakokinetik, Sicherhei tspharmakologie I o t her studies - bioan<llytics. pharmacokinetics and safety pharm<lcology Datu m d e r In spek t io n I C a t e of In spection J u l y 07 - 1 0, 20 0 9 Auf der G r un d lage des lnspek l ionsberichtes und der Besprechun g Ube r zu erfolgende MaBnahmen wird hiermit bosUn i g t , dass in diese r PrOfeinrich t ung die oben genann ten PrOfungen unter Einhaltung der GLP·Grunds.:ltze durchgefOhrt werden k6nne n . I Based on t he inspection report and the discussion of follow up act ivities i t ca n be confirmed , that the test facility is able 1 0 conduct the a forementioned sludies in compliance with the Principles o f GLP

. E i ne Oberprlilung dieser GLP-Bes l a t igun g ist spateslens drei Jahre nach der o . g . lnspe k tion 7.U bean t ragen. D i me diesen Antrag wird nach Ablauf der Frist die P r Qfeinric h lung aus de m dcu l schc n G L P · Obcr w achungsprogramm gcnomme/l und dicse GLP-Best.:ltigung verliert i hre GOltigkcil. I Verification of th is GLP Certificate has 1 0 be applied three years after the above mentioned inspection al the latest. Elapsing this term . the test facility wilt be taken out of the German GlP M onitorin g Programme and this GlP Certificate becomes i nvalid

.

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### Copy No. 2/ 2 

### Page 47 /50

### ANNEXURE 4
. GLP Certificate
- The Netherlands**vocd f)1**  e" wa ren autoriteit


### ENDORSEMENT OF COMPLIANCE WITH THE O!;;CD PRINCIPLES OF GOOD LABORATORY PRACTICE

Pursuant to the Nether l ands GLP Compliance Monitorin g Programme and accord ing to D i rective 2004/9/EC the conformity W iUl the OEeD Principles of GLP was assessed on 4 - 7 May 2009 at

Advinus Therapeutics Priva te Li mited 21 & 22 Phase II , Peenya Industrial Area Bangalore - 560 058 IND I A

It i s herewith confirmed I h at the afo re - mentioned t est facility is currently operating i n co mp l i ance w i th the OECD P rinci p l es of Good Laborato ry P r actice in t h e fOilowing areas of exper1ise: physi ca l·chemical t esting, tox i city stud i es, mutagenicity studies, env ironm enta l studies on aquat i c and terrestrial ani mal s, and analytical ancl clinical chemis try

.

EYE Copy No. 212

### 4'/~**:( i'~L** **~' :'** **:"~~ :**  

## ':.

# 1~**~. -{~** **;{ ,:«** **+h~\"agUe.** 27 August 2 0 09

# ~i**~;i -...,.:.** **:;.~,** ..**~_** ..**:~ :..:,.**  Manager GLP Compl i ance MO flit ori ng Program

Food**d** Con::>um e r Prcduc1 Sil fe fy Auth ori ty (VWA) Prlnses B ea tr j),: J

agn 2

. 2595 A L Den H aag Pcst bus 19 506, 2500 eM De n Haag. The N e theriands



### Page 48/5 0

### ANNEXURE 4 contd
. GLP Ce rtificate
- The Ne therlands

Food and Consume r P roduce Sarety AUlhorit) Mil!isrry 01 A9ricu/rure, Nacure and Food QualilY

Advinus Theri"!peut lCs Limited for the attentio " of Dr. S. Natesan Vice President21 & 22, Phase II, Peenya Industrial Area Bangalo r e • SGO 058 INDI A

Date 18 November 2010 Re. Change of name

Dear Dr. Natesa n, I have received YOllr letter of 26 Octobe r 2010 i n good order. T n yOllr lette r you explained that Advinus Therapeutics Private limited has changed its name into Advinus Therapeutics limited. Furthe r , you stated that the change of name has no consequences fo,' the ownership and that the test facility will continue to function as earlier. The GlP Compliance Monitoring Authority of The Netherlands has periodically inspected Advinus Therapeutics Private Limited

. The most recent inspection wa s carried out on 4 - 7 May 2009
. Subsequent l y, the i nspecto r ate issued an Endorsement of GLP Compliance on 27 A ugust 2009. This endorsement reflects the name of the test fac i lity at t he t i me of the inspection and will no t be renewed u n til a new inspection has been conducted

. Yours sincerely,

Manager GLP Compliance Monitoring Program

EYE Copy No. 212**Pn"~('s ~Jvi** **KIJall** 2 2!J'l) il L Olin**Ha~9** 1' 0 !le x 19 51} 6**25 i1**  eM**D~n** HMg The**Netheri~nd5** ww-" v wa.nl Conl;lct Or. R..M.II. JaSpe'S T +31 {OPO 448 4769 M " 310 f · dl {O)70 449**~747** r**Cb j.JSp("~(~vw~.nl**  Ou r re f

. TR.O
...· . WnI0!293l1

Page 49/ 50

### ANNEXURE 5. GLP Certificate
- India



## NATIO·NAL GLP COMPLIANCE l\fONITORING AUTHORITY
GOOD IAHOIlATORV PRACf1C E 

### GLP CERTIFICATE
GL P Inspectio n was cmric d out a 1 ' Ad\'inus**Thcr:~pcutics** P,"ivatc L imited, Plot Nos
. 2 t & 22, Phase-II , Po s t D o x No
. :;813, J'ccn y a Indu st ri : d Area, Bangal o r..c-S60 058, Inclj" in the following areas 

## or**c;.;pcrt ~c** **:** • ph ys ical
- chemical te s tin g • toxicit y s tudi es • ntu(;lgcnicit ) , s tudie s • cn\'irol1mcntalloxic.ity studies o n aquatic & t er rc s .tri!ll organisms • s tudi es on b e h aviour in water , so il and air, biollCl"Umu lation • , 'cs idu c s tudies • ;\n a"f i.rtical and clinica l chcmisll1' tes tin g**J3:l s~** **d** Oil lhe Inspection Report and th e l ollow-up act io n s take n by the tcst ftlci l ity
. it is co nfirmed that the test faci l ity is
. capahlc of CClIl ducli ng th e ahove·mcnti oncd tc s t sist ud i(: s in co mp l ianc e wi th**OI CD**  Pdllciplc s of Go o d Laboratory Pr ac ti ce (GLP) :lnd Norms, as adop tcd by the**Na ti** **{) ll**  G LP COlllp l inllce M o nitorin g J\u th o rit y. This G LP**C~ 11i** **ficatc** is valid fbI' a pe riod o f three vea l 's frolll Novembe r 7 , 2008 s ubject 10 Ih\.' co ndi ti on that (he les t fa c ili ty compl i es with (h e Terms & Conditions of Ihe Na tion:,1**(~LP** Compl ian ce Monitoring Aut hor j ty 's Document N umber GLP- I01
.

Cc rtificate No.: GLP /C- OI9 I ss ue Datc :



### ·EYE Copy No. 2/ 2 24·12-2008 Head Nat ion :)1 GLP Co mpli:Htce Moltilol'ing Authority Depa rtment of Science & ;rec hn o lo gy**Technolo~** Hlta"
. m New Delhi-I 10016



### Pa ge SO / SO