“INFLUENCE OF VARIOUS ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF PHENOTHIAZINES IN RATS AND MICE.”
M.Pharm Dissertation Protocol
Submitted to the
Rajiv Gandhi University of Health Sciences, Karnataka, BangalorE
by
SANDEEP RAMRAO SURYAWANSHI
Under the Guidance of
SRI.ASHOK M.DASTAPUR
M.Pharm.
Department of Pharmacology
H.K.E. Society’s College of Pharmacy,
Gulbarga-585 105
2008-09
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES KARNATAKA, BANGALORE
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidate andAddress (in block letters) / SANDEEP RAMRAO SURYAWANSHI
SHIVKALYAN NAGAR,
A/P: LOHA, TAL: LOHA, DIST: NANDED, 431708.
2. / Name of the Institution / H.K.E. Society’s College of Pharmacy,
Sedam Road, GULBARGA-585 105
3. / Course of the study
Branch / M. Pharm.,
Pharmacology
4. / Date of Admission to Course / 1/06/2008
5. / Title of the Research topic / INFLUENCE OF VARIOUS ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF PHENOTHIAZINES IN RATS AND MICE.
6. / Brief Resume Of Intended Work
6.1 Need for the study
/ Enclosure-I6.2 Review of Literature
/ Enclosure-II6.3 Objective of the study / Enclosure-III
7. / Materials and Methods
7.1 Source of data / Enclosure-IV
7.2 Methods of collection of data / Enclosure-V
7.3 Does the study require any
Investigation on animals?
If yes give details / Enclosure-VI
7.4 Has the ethical clearance been
obtained from your institution in
case of 7.3 / Yes, Regsitration number :142/1999 CPCSEA.
5 th JULY 1999
8. / List of References / Enclosure-VII
9. / Signature of the candidate / (Sandeep Ramrao Suryawanshi)
10. / Remarks of the guide / Enclosure -VIII
11. / Name and Designation of
(in Block Letters)
11.1 Guide
11.2 Signature
11.3 Co-Guide (if any)
11.4 Signature
11.5 Head of the department
11.6 Signature / Sri. ASHOK M.DASTAPUR,
Asst. Professor,M. Pharm
Dr. NITIN MAHURKAR, Professor, M. pharm., PhD; EDPCM.
Dr. K.PRASAD
Professor,
M. pharm. PhD
12. / Remarks of the Principal
12.1 Signature / The present study was permitted for executing the work in the institution and IAEC permission has been granted.
(Dr. Appala Raju)
Enclosure-I
6. Brief Resume on the Intended Work
6.1 Need for the Study:
Phenothiazine derivative like chlorpromazine are used to depress CNS and they have sedative and tranquilizing properties. They are used for the management of psychotic disorders and act by inhibiting the release of dopamine.2,14
The commonly used phenothiazines are classified based on the structure which,
1] Possess Aliphatic side chain: Chlorpromazine, Triflupromazine
2] Possess Piperidine side chain: Thioridazine
3]Possess Piperazine side chain: Trifluoperazine , Fluphenazine
Therapy with phenothiazines invariably causes extrapyramidal symptoms as a major dose limiting side effect. They are more prominent with high potency drugs like fluphenazine, haloperidol and least with thioridazine, clozapine, and the symptoms include Parkinsonism, acute muscular dystonias, akathesias, tardive dyskinesia, malignant neuroleptic syndrome.
Extrapyramidal symptoms in Parkinsonism are associated with rigidity, tremor, hypokinesia, mask like facies appears between 1-4 weeks of therapy alogwith postural abnormilities and bradykinesia, etc.1
An antioxidant is a molecule that is capable of slowing or preventing the oxidation of other molecules. Oxidation reactions can produce free radicals which starts chain reaction that damage cells.
Oxidation reactions are crucial for life, they can also be damaging hence plants and animals maintain complex system of multiple types of antioxidants such as glutathione, vit. C and E as well as enzymes such as catalase, super oxide dismutase and various peroxidases. Low level of antioxidants or inhibition of antioxidant enzymes causes oxidative stress and may damage or kill cells.
Oxidative stress has a role in cellular damage and contributes to pathophysiology of many diseases. This has a role in autoimmune disorders, jaundice, cancer, atherosclerosis etc.
Antioxidants commonly contain a mixture of β-carotene, vitamine C and Vitamine E, minerals, coenzyme Q etc, which have synergistic antioxidant effects and it effectively terminate free radical activity implemented in tissue damage.3,4,9
Antioxidant enzyme super oxide dismutase (SOD) catalase and glutathione peroxidase help to catalyse the reduction of oxidants in a cell and exert their effects by counteracting oxidative processes that contribute to the cause of the chronic diseases.6,7,8
The literature also reveals that phenothiazines decrease the level of coenzyme Q that may further aggrevate extra pyramidal symptoms. Hence in the present project it is proposed to study the influence of antioxidants along with phenothiazines so as to suggest a safe therapy as both the drugs are widely used already clinically.
Enclosure-II
6.2 Review of literature
An extensive literature review has been carried out to find out whether similar work has been reported elsewhere and review indicates no similar reports.
The following is a brief report of literature-11
1)Phenothazine depress the level of antioxidants.
2)Antioxidant have beneficial effect in controlling extrapyramidal symptoms.
3)Antioxidants acts by free radical scavenging mechanism.
1)Chopra, R., Goldman, R., Bhagwan, H.N.,Relative bioavailability of Q10 formulations. Annual Meeting Abstracts 108;J Amer Ph 1998;Vol38,No.2.
2)Baggio E.,Gandini R., Plancher A.C.,Passeri M.,Carmosino G(1994): Italian multicenter study on the safety and efficacy of coenzyme Q10 as adjunctive therapy in heart failure,CoQ10 drug surveillance investigators.Molecular Aspects of Medicine,15:287-294.
3)Bhagavan H. N., Chopra R. K. (2006) :Coenzyme Q10:Absorption ,tissue uptake, metabolism pharmacokinetics.Free Radical Research,40:445-453.
Enclosure-III
6.3 Objectives of Study:
The study is intended to assess the role of antioxidants in Parkinsons therapy since Parkinsons is an autoimmune disorder with no cure. Requiring life long treatment, the present study is intended,
1)To evaluate and recommended a safe combination.
2)To reduce dose of antiparkinsonian drugs.
3)To control the disorder more smoothly and effectively.
4)To reduce toxicity and side effects of phenothiazines especially extrapyramidal symptoms
Enclosure-IV
7.1 Source and Data
-H.K.E’s library.
-Medline net.
-Del net,Helinet,Pubmed.
-Pharmacological reviews.
Data collected from albino rats and mice produced from central animal house M.R.medical college, Gulbarga.
Phenothiazine pure sample will be collected from reputed manufactures.
Antioxidants will be produced from reputed manufactures.
Enclosure –V
7.2 Method of Collection of Data
The study is planned on normal rats and mice and work is designed in following phases:
Phase-I In this phase it is proposed to establish standard dose response of phenotiazines for extrapyramidal symptoms in rats and mice in our laboratory.
Phase-II This phase involves evaluation of extrapyramidal symptoms by administering combination of phenothiazines and antioxidants in rats.
Phase-III This phase involves evaluation of extrapyramidal symptoms by administering combination of phenothiazines and antioxidants in mice.
Inclusion Criteria-
- Normal Wistar albino rats weighing between 180-220 gms.
- Normal Wistar albino mice weighing between 25-30 gms.
Exclusion Criteria-
- Albino rats which do not fall in the above weight range are excluded from the study.
- Albino mice which do not fall in the above weight range are excluded from the study.
Study Sampling:
- The response is evaluated by using standard probs.
- Statistical significance is calculated by standard T-test.
- The total duration of study extends up to 10 months.
Enclosure- VI
7.3 Does the study require any investigation on animals? If so, please describe briefly-
Yes, the above study requires investigation on animals that is albino rats and mice for extra pyramidal activity.
7.4 Has the ethical clearance has been obtained from your institution in case 7.3.
Yes, the study is cleared from institutional animal ethics committee [IAEC Certificate enclosed]
Enclosure-VI
Application for Permission for animal experiments.
Application to be sumitted to send either to the cpcsea (address in form a above) or institutional animal ethic committee (iaec).
part A
1 / Name and Address of Establishment / H.K.E.S’s College of Pharmacy, Sedam Road, GULBARGA. karnataka- 585 1052 / Registration Number and Date of registration. / 142/1999 CPCSEA. 5th jULY 1999
3 / Name, address and Registration number of breeder from whom animal acquired (or to be acquired) for Experiments mentioned in part B and C. / cENTRAL ANIMAL HOUSE
M.R.MEDICAL College, sedam road
gulbarga-585 105 kARNATAKA
142/1999 CPCSEA. 5th jULY 1999
4 / Place where the animals are presently kept (or proposed to be kept) / cENTRAL ANIMAL HOUSE
M.R.MEDICAL College, sedam road
gulbarga-585 105 kARNATAKA
5 / Place where experiment is to be performed / pharmacology & toxicology laboratory. post graduate department H.K.E.S’s College of Pharmacy-GULBARGA
6 / Date on which the experiments is to commence and duration of Experiment. / 01.06.2009
10 months
(the appropriate protocol form for the research proposal part-b in the case experiments using animals other than on human pRimates, part-c for use of non-human primates to be duly filleD in. signed and aTTACHED to this form).
Date :signature
Place :
Name and designation of Chief Investigator
Sri. Ashok Dastapur ,M.pharm,
Asst.Professor, Deparment of Pharmacology
H.K.E.S’s College of Pharmacy,
Sedam Road GULBARGA.
karnataka- 585 105
Animal experiment for research purpose
1 / Name and Address / sANDEEP RAMRAO SURYAWANSHI.SHIVKALYAN NAGAR, a/p.LOHA, tQ:LOHA dist: NANDED, maharashtra-431708.
2 / Area of Research / Pharmacology
3 / List of Experiments Carried out / INFLEUNCE OF VARIOUS ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF PHENOTHIAZINEs IN RATS AND MICE
4 / No. of animals used annually (species-wise) / 60 Rats
60 Mice
5 / Duration of the Project / 10 Months
6 / Source of Experimental animals (In-house Breeding/other Institution/import/others) / cENTRAL ANIMAL HOUSE
M.R.MEDICAL College, SEDAM road
gulbarga-585 105 kARNATAKA
7 / No. of Animals to be sacrificed / NIL
8 / Method of euthanasia and disposal / The Euthanasia will be done under mild ether anesthesia in rats
9 / Whether a Veterinary doctor is employed / yes
10 / Housing details (Dimension/No. of animals) / 254 Sq.m.
11 / No. of animals rehabilitated / ----
12 / In vitro research / ----
13 / Scientific findings (update when project is over) / INFLEUNCE OF VARIOUS ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF PHENOTHIAZINE in RAts AND MICE
Kindly fill in the appropriate from-research / Educational details to be precise.
Signature of the IAEC Chairman,
Part – b
Protocol forms for research proposals to be submitted to the committee / Institutional Animal Ethics Committee, for new experiments or extensions of ongoing experiments using animals other non-primates.
1 / Project Title / INFLEUNCE OF VARIOUS ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF PHENOTHIAZINEs IN RATS AND MICE2 / Chief Investigator :
- Name :
- Designation :
- Dept/Div/Lab :
- Telephone number :
Asst. Professor
Dept. Of Pharmacology
H.K.E.S’s College of Pharmacy, Sedam Road GULBARGA.585 105 kARNATAKA
(08472)-221392
9900217179
3 / List of names of all individuals authorized to conduct procedures under this proposal. / Sri. Ashok Dastapur
Dr. K. Prasad
Sri. Neelakanth Reddy
Dr.Nitin Mahurkar
Mr. Sandeep Suryawanshi
4 / Funding source / H.K.E.S’s. college of pharmacy,
GULBARGA-585 105.
5 / Duration of the project :
- Number of months :
- Date of initiation :
- Date of completion :
01.06.2009
31.03.2010
6 / If day by which approval is needed is less than six weeks from date of submission. Justification for the same. / Doesn’t arise.
7 / Study objectives (the aims of study and why they are important) to be explained non-technical terms as for as possible. / INFLEUNCE OF VARIOUS ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF PHENOTHIAZINEs IN RATS AND MICE
8 / Animals required
- Species
- Age/weight/size
d. Number to be used
- Number of days each animal will be housed
Adult Rats –180-220gm
Adult Mice -25-30gm
Either sex
Rats 60
Mice 60
45 Days.
9 / Rationale for animals usage :
- Why is animal necessary for these studies?
- Why are the particular species selected?
- Why is the estimated number of animals essential?
- Similar experiments conducted in the past. If so, number animals used and results obtained in brief.
- If yes, why new experiment I required?
- Have similar experiment (s) been made by any other organization/agency? If so, their result in your knowledge.
These species are suitable for screening extrapyramidal symptoms as they possess the complications that are comparable to human beings.
To meet minimal statistical requirements
- No. -
- No. -
- No. -
10 / Does the protocol prohibit use of anesthetic or analgesic for the conduct of painful procedures? / - No. -
11 / Will survival surgery be done? / - No. -
12 / Methods of disposal of post experimental Animals / Similar to Sl. No. 8 as in 1st page
13 / Animal transportation methods if extra institutional transport is envisaged. / - No. -
14 / Use of hazardous agents / - No. -
Investigator’s declaration.
1)I certify that I have determined that the research proposal here in is not unnecessarily 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 procedures.
3)For procedure listed under item 11, have reviewed the pertinent scientific literature and have found no valid alternative to any procedure described here in which may cause less pain or distress.
4)I will obtain approval from the iaec/cpcsea before initiating any significant changes in study.
5)Certified that performance of experiment will be initiated only upon review and approval of scientific intent by appropriate expert body (to be named).
6)Institutional bio-safety committees (IBC) Certification of review and concurrence will be taken (required for studies utilizing DNA Agent of human pathogens.
7)I shall maintain all the records as per format (form-D).
Date :Signature
Place:
Name and designation of Chief Investigator
SRI. ASHOK DASTAPUR , M. pharm.
Asst. Professor, Department of Pharmacology,
H.K.E.S’s College of Pharmacy,
Sedam Road GULBARGA.
karnataka- 585 105
name / signatureSRI. ASHOK M. DASTAPUR,
Asst. Professor (Guide)
Dr. Nitin Mahurkar – Professor,
(Co-Guide)
Dr. K. Prasad-Professor
Sri. Neelakanth Reddy Asst. Professor
Mr. Sandeep Suryawanshi-Student
(For iaec/cpcsea Usage)
Proposal Number: HKE COP/ IAEC / 02 / 2008-09
Date first received: 12/11/08
Date received after modification (if any): ------
Approval Date: 12/11/08
Expiry Date: 31/03/2010
Name of IAEC/CPCSEA Chair person: Dr.S.A. Raju
Principal, HKE’S COP,
Gulbarga-585105
Enclosure-VII
List of References:
1) Goodman and Gilman’s, The Pharmacological Basis Of Therapeutics 10th Ed. McGraw Hill, Medical Publication Division, New York, 2006;552-560.
2) Rang H. P., Dale M. M., Ritter J. M., Moore P. K. Pharmacology 2003; 497-501, 373-374.
3) Oberiey LM. Free radicals and diabetes. Free radicals Bio Med 1988; 5:113-124.
4)Buscigllo J, Yankner BA. Apoptosis and increased generation of reactive oxygen species in Downs syndrome neurons in vitro nature. 1995; 378: 776-779.
5)Maurice MM, Nabamura H, Van Der voort EAM, Van Vliet Ai, Staal FJT, Tak PP, Breedveld PC, Verweij CL. Evidence for the role of an altered redox state hyporesponsiveness of synovial T-cells in rheumatoid arthritis J Immunol 1997; 1458-1465.
6)Evans PH. Free radicals in brain metabolism and pathology. In: Cheeseman KH., Slater TF editors. Free radicals in medicine. Churchill Livingstone, London, 1992; 577-587.
7)Halliwel B, Gutteridge JMC, Carrol EC. Free radicals antioxidants and human diseases: where are we now? Lab din med 1994; 119: 598-619.
8)Mittal MM. Antioxidant medicine update 1999; 9: 205-212.
9)Diplock AT, Machlin LJ, Packer L, Pryor WA. Vitamin E; Biochemistry health Implications. Ann NY Acad Sci 1989; 570: 555-590.
10)Asada K. Ascorbate peroxides hydrogen peroxide- scavenging enzyme in plants. Physiol plant 1992; 85: 235-241.
11)Chopra, R., Goldman, R., Bhagwan, H.N.,Relative bioavailability of Q10 formulations. Annual Meeting Abstracts 108;J Amer Ph 1998;Vol38,No.2.
12)Baggio E.,Gandini R., Plancher A.C.,Passeri M.,Carmosino G(1994): Italian multicenter study on the safety and efficacy of coenzyme Q10 as adjunctive therapy in heart failure,CoQ10 drug surveillance investigators.Molecular Aspects of Medicine,15:287-294.
13)Bhagavan H. N., Chopra R. K. (2006) :Coenzyme Q10:Absorption ,tissue uptake, metabolism pharmacokinetics.Free Radical Research,40:445-453.
14)Essentials of Medical Pharmacology by K.D.Tripathi ;6 th Ed.,423-433.
Enclosure-VIII
Remark of the guide:
The above work is original and genuine and is feasible in our laboratory, hence recommended for favour of consideration.
HKECOP/IAEC/02/2008-09