“INFLUENCE OF Selective antioxidants on extrapyramidalsymptoms of selected
phenothiazines in rats”
M.Pharm Dissertation Protocol
Submitted to the
RajivGandhiUniversity of Health Sciences,Karnataka, BangalorE
by
ZOHARA FATIMA KHATIB
UNDER THE GUIDANCE OF
MR.ASHOK KUMAR M. DASTAPUR
M.Pharm
Department of Pharmacology
H.K.E. Society’s College of Pharmacy,
GULBARGA-585 105
2010-2011
RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES KARNATAKA, BANGALORE
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidate andAddress (in block letters) / ZOHARA FATIMA KHATIB
H.No-2-907/121/58 Near LuqmanCollege of Science Badepur Layout ,Gulbarga-585105
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 / 15/06/2010
5. / Title of the Research topic / InfluenceOF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF SELECTED PHENOTHIAZINES IN RATS.
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, Registration number: 142/1999 CPCSEA.5th July 1999.
8. / List of References / Enclosure-VII
9. / Signature of the candidate / (Zohara Fatima Khatib)
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 / MR.ASHOK KUMAR M.DASTAPUR
M.PHARM
ASST.PROFESSOR
DR.NITIN MAHURKAR
M.PHARM.,PhD
PROFESSOR
Dr.NITIN MAHURKAR
M.PHARM., PhD
Professor
12. / Remarks of the Principal
12.1 Signature / The present study was permitted for executing in the institution and IAEC permission has been granted.
(Dr. S. Appala Raju)
ENCLOSURE-I
6. Brief Resume on the Intended Work
6.1 Need of the Study:
Phenothiazine derivatives like chlorpromazine,triflupromazine,thioridazine are used to depress CNS and they are sedative and have tranquilizing properties. They are used for management of psychotic disorders and actby inhibiting the release of dopamine.1, 2
The commonly used phenothiazines are chlorpromazine, triflupromazine, thioridazine, trifluperazine, fluphenazine etc.
Therapy with phenothiazines invariably causes extra pyramidal symptoms as major dose limiting side effect. They are more prominent with high potency drugs likefluphenazine, haloperidol and least with thioridazine, clozapine and the symptoms includeparkinsonism, acute muscular dystonias, akathesias, tardive dyskinesia, malignant neuroleptic syndrome.
Extrapyramidal symptoms in parkinsonism are associated with rigidity, tremor,hypokinesia, mask like facies appear between 1-4 weeks of therapy along with postural abnormalities and bradykinesia etc.1
Antioxidants are substances which help defend the body against cell damage caused by various free radicals.3
Antioxidant enzyme superoxide dismutase (SOD),catalase and glutathione peroxidase help tocatalyse the reduction of oxidants in a cell and exert their effect by counteracting oxidative process that contribute to the cause of the chronic diseases.4, 5, 6
ENCLOSURE-II
6.2 Review of literature
An extensive literature review has been carried out to find whether similar work hasbeen reported elsewhere and review indicates no similar reports.
The following is a brief report of literature
- VitaminC, E and beta carotene are well known antioxidants3
- Selenium is a trace mineral and potent antioxidant. Which are primarily involved in antioxidant function and redoxstatus. It is widely distributed throughout the body but is particularly well maintained in the brain. Changes in selenium concentration in blood and brain have been reported in Alzheimer’s disease and brain tumors.7, 10
- Curcumin (turmeric) an ancient Indian herb used as curry powder, has been extensively studied in modern medicine and Indian system of medicine for the treatment of various medical conditions including cystic fibrosis, haemorrhoids, gastriculcer, coloncancer, breastcancer, atherosclerosis.
It has been used in various types of treatment for dementia and traumatic brain injury.Curcumin also has a potential role in prevention and treatment of Alzheimer’s disease.Curcumin as an antioxidant and anti-inflammatory and lipophilic action improves the cognitive function in patients with Alzheimer’s disease.8
- KulkarniSK et al have reported the potentials of curcumin as an antidepressant and in treatment of various central nervous system disorder.9
- Brauer Au has reported the neuroprotective mechanism of selenium.10
- Jyothi A et al have reported the protective effect of curcumin against electro behavioural progression of seizures in the iron-induced experimental model of epileptogenesis due to its ability to cross blood-brain barrier and bind redox-active metal ions. 11
- Saito Y et al have reported the cell death caused by selenium deficiency and protective effect of antioxidants.The results suggests that the reactive oxygen species,especially lipid hydroperoxides are involved in the cell death caused by selenium deficiency and that selenium and vitamin E co-operate in the defense against oxidative stress upon cells by detoxifying and inhibiting the formation of lipid hydroperoxides.12
Enclosure-III
6.3 Objective of Study:
The study is intended to assess the role of selective antioxidants [Selenium, Curcumin the active principle of turmeric (Curcuma Longa) family: Zingiberaceae] on extrapyramidal symptoms induced by selective phenothiazines.
The present study is planned with the following objectives.
- To reduce toxicity and side effect of phenothiazines especially extrapyramidal symptoms.
- To evaluate and recommend a safe combination.
- To control the disorder more smoothly and effectively.
PARAMETERS:
- Superoxide radical scavenging activity.
- Hydroxyl radical scavenging activity.
- Lipid peroxidation inhibitionactivity.
Enclosure-IV
Materials and Methods
Source and Data
This study is planned to generate data by conducting laboratory based research in animals. It is also planned to use rats in the experiment.
Other data is collected from
- Pharmacology reviews, journals.
- Electronic data like CD-ROM and internet.
- Helinet, Delnet, pubmed.
- Library of H.K.E’s college of pharmacy, Gulbarga.
The animals will be procured from central animal house, MRMedicalCollege, Gulbarga.
Phenothiazine (chlorpromazine) pure sample will be collected from reputed manufacturers.
Antioxidants will be procured from reputed manufacturers.
Enclosure –V
Method of Collection of Data
The study is planned on normal rats and work is designed in following phases.
Phase I: In this phase it is proposed to establish standard dose response of phenothiazines for extrapyramidal symptoms in rats in our laboratory
Phase II: This phase involves evaluation of extra pyramidal symptoms by administering combination of phenothiazines and selected antioxidants (Selenium, Curcumin) in rats.
Inclusion Criteria
Normal Wistar albino rats weighing between 180-220 gms.
Exclusion Criteria
The albino rats which do not fall in the above weight range are excluded from the study.
Study Sampling
1)The response is evaluated by using standard probes.
2)Statistical significance is calculated by student t test.
3)The total duration of experimentation will be 10 months.
Enclosure –VI
7.3 Does the study require any investigation on animals? If so, please prescribe briefly
Yes, the above study requires investigation on animals that is albino rats for extrapyramidal activity.
7.4 Has the ethical clearance been obtained from your institution in case of 7.3
Yes, the study is cleared from institutional animal ethics committee (IAEC Certificate enclosed).
Enclosure-VII
List of References
- Goodman and Gilman`s. The Pharmacological Basis of Therapeutics McGraw Hill, MedicalPublication Division, New York, 10thEd. 2006.
- Rang HP, Dale MM, Ritter JM, Moore PK.Pharmacology 5th Ed.Edinburg: Churchill Livingstone; 2003.
- Dhrubo Jyoti Sen. HerbalAntioxidants: A great hope for future.Pharma Times.2008;40(12).
- Diplock AT, Machlin LJ, Packer L, PryorWA.VitaminE; Biochemistry health implication. Ann NY Acad Sci 1989;570:555-90.
- Chopra R, Goldman R, Bhagwan HN.Relative bioavailability of Q10 formulation. Annual Meeting Abstracts 108; J Amer Ph 1998;38(2).
- Tripati KD Essentials of Medical Pharmacology.6thEd.New Delhi: Jaypee Brothers Medical Publisher (P) Ltd; 2008.
- Chen J, Berry MJ. Selenium and selenoproteins in the brain and brain diseases.Journal of Neurochemistry. 2003 Jul; 86 (1):1-12.
- Mishra S, Palanivelu K.The effect of curcumin(turmeric)in Alzheimer’s disease. Annals of IndianAcademy of Neurology.2008 Jan; 11(1):13-9
- KulkarniSK, Dhir A, Akula KK.Potentials of curcumin as an antidepressant. Scientific World Journal. 2009; 9:1233-41
- Brauer Au,Savaskan NE.Molecular action of selenium in the brain: neuroprotective mechanism of an essential trace element. Reviews in the Neurosciences.2004; 15(1):19-32.
- Jyoti A, Sethi P, Sharma D. Curcumin protection against electrobehavioural progression of seizures in the iron-induced experimental model of epileptogenesis. Epilepsy Behav 2009;14:300-8.
- Saito Y, Yoshida Y,Akazawa T,Takahashi K, Niki E.Cell death caused by selenium deficiency and protective effect of antioxidants.Journals of Biological Chemistry 2003 Oct 10;278 (41):39428-34.
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.MEDICALCollege, 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.MEDICALCollege, 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. / 01.06.2011 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
Mr.ASHOK KUMAR M.DASTAPUR
M.pharm
Asst.Professor,
H.K.E.S’s College of Pharmacy,
Sedam RoadGULBARGA.
karnataka- 585 105
Animal experiment for research purpose
1 / Name and Address / ZOHARA FATIMA KHATIBH.NO-2-907/121/58 NEAR LUQMANCOLLEGE OF SCIENCE ,BADEPUR LAYOUT GULBARGA- 585105
2 / Area of Research / Pharmacology
3 / List of Experiments Carried out / influenceOF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF SELECTED PHENOTHIAZINES IN RATS.
4 / No. of animals used annually (species-wise) / 60 Rats
5 / Duration of the Project / 10 Months
6 / Source of Experimental animals (In-house Breeding/other Institution/import/others) / cENTRAL ANIMAL HOUSE
M.R.MEDICALCollege, SEDAM road
gulbarga-585 105 kARNATAKA
7 / No. of Animals to be sacrificed / No sacrifice
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) / InfluenceOF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF SELECTED PHENOTHIAZINES IN RATS
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 / InfluenceOF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF SELECTED PHENOTHIAZINES IN RATS.2 / Chief Investigator :
- Name :
- Designation :
- Dept/Div/Lab :
- Telephone number :
Asst.Professor
Dept. Of Pharmacology
H.K.E.s`S College of Pharmacy,Sedam RoadGULBARGA - 585 105 kARNATAKA
09900217179
3 / List of names of all individuals authorized to conduct procedures under this proposal. / Mr.ASHOK KUMAR M.DASTAPUR
Dr. NITIN MAHURKAR
ZOHARA FATIMA KHATIB
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 :
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. / InfluenceOF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF SELECTED PHENOTHIAZINES IN RATS.
8 / Animals required
- Species
- Age/weight/size
- Gender
- Number to be used
- Number of days each animal will be housed
Rats 60 40 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 disorders of 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
Mr.ASHOK KUMAR M.DASTAPUR
M.pharm
Asst.Professor,
H.K.E.S’s College of Pharmacy,
Sedam RoadGULBARGA-585105
karnataka.
name / signatureMR.ASHOK KUMAR M.DASTAPUR (Guide)
ZOHARA FATIMA KHATIB-Student
.
(For iaec/cpcsea Usage)
Proposal Number: HKE COP / IAEC / 36 / 2010- 2011
Date first received:25-11-2010
Date received after modification (if any): ----
Approval Date:06-12-2010
Expiry Date:30-04-2012
Name of IAEC/CPCSEA Chair person: Dr. S. Appala Raju.
Principal,
H.K.E. Society’s College of Pharmacy,
Gulbarga.585105
Enclosure-VIII
Remark of the guide
The above work is genuine and original and is feasible in our laboratory, hence recommended for favour of consideration.
HKE COP/IAEC/36/2010-2011