Part B Protocol form for research proposal to be submitted to the institutional animal ethical committee for conduct of experiment

/ extensions of ongoing experiments on animals other than non-human primate. 1. Project Title To study pharmacokinetic parameter of Solid Dispersion system of Anti-HIV drug.

2. Chief investigator : a. Name b. Designation c. Dept./Div./Lab. d. Mobile no. e. Name of student f. Purpose of study : Mr. V. R. Katariya : Lecturer : 7588930701 [Mr. V. R. Katariya] : Ms. Savita B. Patil : A Part of Post graduate research work

3. List of all individuals authorized to conduct procedure under this proposal :, Mr. V. R. Katariya, Dr.R.B.Nawale, Savita Patil, Priya Shinde.

4. Funding source

: Self

5. Duration of project a. Date of initiation

2 year : Animal studies will start soon after ethical committees approval. : 2 year

b. Date of completion

6. If date by which approval is needed is less than 6 weeks from the date of submission, justification for the same : Not Applicable

7. Study objectives 8. Animal required Animal Species Weight Age No. To be used Gender

To study pharmacokinetic parameters. Statistical evaluation of data. Rats Albino Wistar Rats 250-300 gm Young & Healthy 18 nos. Either sex Throughout experimentation in the lab.

Rabbits New Zealand White Rabbit 1.5-2.0 Kg Young & Healthy 06 nos. Either sex

b. Number of days each animal will be housed :

9. Rational for animal usage: a. Why animal usage necessary for the study From literature survey it reveals that the rabbits GIT used as alternative to human GIT to determine the pharmacokinetic parameters. Literature survey shows that rabbits GIT closely resembles with human system and hence can be used for In-vivo studies. The estimated numbers of animals are required to carry out well designed statistical experiments. : No : Not applicable : Not applicable

b. Why are the particular species selected required

c. Why is the estimated number of animals essential

d. Similar experiments conducted in our laboratory in past e. If yes why experiment is required f. Have similar experiments been made by any other organization/agency, if so their results in your knowledge 10. Description of procedure to be used

The animals will be brought from authentic source. They will be kept in college animal house under standard conditions during and after the experimentation till their

presence in the college. They will be feed with standard food obtained from authentic source. Water will be provided to animal ad libindum On the day of experimentation the gross health of animal will be observed. Animal will be fastened overnight before feeding of formulation sample. The formulation will be feed orally with the help of oral feeding cannulae. The blood sample withdrawn at interval of each one hour for 12 hours from lateral saphenous vein. The blood sample will centrifuge, separate the plasma. Extraction of the drug from blood sample carried out & concentration of drug will determined by the pre developed HPLC method. The bioavailability of drug will be determined.

References: 1.Bregt S Kappelhoff1*, Frank van Leth2, Thomas R MacGregor3, Joep MA Lange2, Jos H Beijnen Nevirapine and efavirenz pharmacokinetics and covariate analysis in the 2NN study. 2.Peter Langmann*, Diana Schirmer, Thomas Vath, Steffen Desch, Michael Zilly, Rapid determination of nevirapine in human plasma by gas chromatography. 11. Does the protocol prohibit the use of anaesthetic and analgesics for the : NO conduct of painful procedures: 12. Will survival surgery be done 13. Method of disposal post experimentation : No. : Not Applicable No

14. Animal transportation method if extra-institutional transport is envisaged: 15. Use of hazardous agent:

No

UNDERTAKING I / we undertake that will obey all the guidelines laid down by CPCSEA for the use of animals for experimentation purpose. will maintain the animals as per guideline of the CPCSEA and will not ask for any reimbursement from institute for the same. will inform well in advance to the officials of IAEC before bringing the animals to the institute. will bring the animal from legal source only (CPCSEA approved suppliers) will inform immediately to the officials of IAEC after bringing the animals to the institute. will provide all the information necessary for official maintenance of the record to the institute and will not object for that. I will not transport the animals extra institutional if not permitted to do so (as per point no. 14). I will implement all the procedures as per given above and any change I will bring in advance to the officials of IAEC. I / We all who have authorized for the experimentation have taken training for the procedure to be executed for the animal use.

Signature with name of all the persons related to this approval

Guide : Mr. V. R. Katariya

Student1 : Savita Patil

2: Priya Shinde

INVESTIGATORS DECLARATION 1. I certify that I have determined that the research proposal herein is not unnecessary duplicative of previously reported research. 2. I certify that all individuals working on this proposal and experimenting on the animals have been trained in animal handling procedure. 3. For procedures listed under item 10, I certify that I have review the pertinent scientific literature and have found no valid alternative to any procedure described herein which may cause less pain or distress. 4. I will obtain approval from the IAEC/CPCSEA before initiating any significant changes in this study. 5. Certified that performance of experiment will initiate only upon review and approval of scientific intent by appropriate scientific expert body (Institutional scientific advisory committee/funding agency/ other body). 6. Institutional biosafety committees (IBC) certification of review and concurrence will be taken (required for studies utilizing DNA agent of human pathogen). 7. I shall maintain all the record as per format (form D).

Signature Date: 3/3/2012 Name: Mr. V. R. Katariya

------------------------------------------------------------------------------------------------------------------------------------(For IAEC/CPCSEA usage) Proposal number Date first received Date received after modification (if any) Date received after second modification (if any) Approval date Expiry date Name of IAEC/CPCSEA chairperson

Date:

Signature