6. Brief Resume on the Intended Work : 6. 1 Need for the Study : Phenothiazine derivative like chlorpromazine are used to depress CNS and they are sedative and tranquilizing properties. They are used for the management of psychotic disorders and act by inhibiting the release of dopamine 1,2 . The commonly used phenothiazines are chlorpromazine, triflupromazine, thioridazine trifluperazine , fluphenazine etc. 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 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 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 4,5,6. 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 3 1. Vitamin C and E and beta carotene are well known antioxidants. 2.
Vitamin E is only moderately effective against single oxygen, it is the most effective antioxidant for terminating the chain reactions of lipid peroxidation in call membranes. 3. CoQ is a part of antioxidant defense network that protects lipids from free radical attacks. ? Mahlberg et al have reported melatonin treatment of Day- Night Rhythm disturbances and sundowning in Alzheimer disease 7. ? Reiter RJ et al have reported melatonin has been shown to reduce the toxicity and increase the efficacy of a large number of drugs whose side effects are well documented 8.
? Meena Shrivastava et al have reported relapse of tardive dyskinesia due to reduction in clozapine dose 9. ENCLOSURE – III Objectives of the study: The study is intended to assess the role of selective antioxidants (melatonin, vitamin E) on extrapyramidal symptoms induced by selected phenothiazines. The present study is planned with the following objectives. 1. To reduce toxicity and side effects of phenothiazines especially extrapyramidal symptoms. 2. To evaluate and recommended a safe combination. 3. To control the disorder more smoothly and effectively. Parameters: 1.
Superoxide radical scavenging activity. 2. Hydroxyl radical scavenging activity. 3. Lipid peroxidation inhibition activity. ENCLOSURE – IV Materials and Methods Source of Data This study is planned to generate data by conducting laboratory–based research in animals. It is also planned to use rats in the experiments. 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, MR Medical college, Gulbarga.
? Phenothiazine (chlorpromazine) pure sample will be collected from reputed manufacturers. ? Antioxidants will be produced 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 extrapyramidal symptoms by administering combination of phenothiazines and selective antioxidants (melatonin, vitamin E) in rats.
Inclusion Criteria- Normal Wistar albino rats weighing between 180-220 gms. Exclusion Criteria- 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 probs. 2. Statistical significance is calculated by standard t-test. 3. 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 has 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 REFERENCES 1. Goodman and Gilman`s, The Pharmacological Basis Of Therapeutics McGraw Hill, Medical Publication Division, New York, 10th Ed. 2006. 2. Rang HP, Dale MM, Ritter JM, Moore PK. Pharmacology 5th Ed. Edinburg: Churchill Livingstone; 2003. 3. Prof. (Dr. ) Dhrubo Jyoti Sen. Herbal Antioxidants: A great hope for future. Pharma Times. 2008; 40(12). 4. Diplock AT, Machlin LJ, Packer L, Pryor WA.
Vitamin E; Biochemistry health implications. Ann NY Acad Sci 1989;570: 555-90. 5. Chopra R, Goldman R, Bhagwan HN. Relative bioavailability of Q10 formulations. Annual Meeting Abstracts 108;J Amer Ph 1998; 38(2). 6. Tripathi KD Essentials of Medical Pharmacology. 6th Ed. New Delhi: Jaypee Brothers Medical Publisher (P) Ltd; 2008. 7. Mahlberg, Richard MD, Kunz, Dieter MD, Sutej, Igor MD et al. Melatonin treatment of day night rhythm disturbance and sundowning jn Alzheimer disease. Journal of Clinical Psychopharmacology. 2004; 24(4) : 456-59. 8. Reiter RJ, Tan DX, Sainz RM, Mayo JC, Lopez Borillo S.
Melatonin: Reducing the toxicity and increasing the efficacy of drugs. Journal of Pharmacy and Pharmaccolgy. 2002; 54(10): 1299-1321. 9. Meena Shrivastava, Bhopendra Solanke, Ganesh Dakhale, Abhishek Somani, Pravir waradkar. relapse of tardive dyskinesia due to reduction in clozapine dose. Indian journal of Pharmacology. 2009; 41(4). Application for Permission for animal experiments. Application to be suBmitted 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| | | |105 | |2 |Registration Number and Date of registration. |142/1999 CPCSEA. 5th jULY 1999 | |3 |Name, address and Registration number of breeder from whom animal |cENTRAL ANIMAL HOUSE | | |acquired (or to be acquired) for Experiments mentioned in part B |M. R. MEDICAL College, sedam road | | |and C. |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 |01. 06. 2010 | | |
Experiment. |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 appended to this form). Date :signature Place : Name and designation of Chief Investigator Dr. K. PRASAD, M.
PHARM. PhD, Professor, H. K. E. S’s College of Pharmacy, Sedam Road, GULBARGA. karnataka- 585 105 Animal experiment for research purpose |1 |Name and Address |SANTOSH RAMESH AWASTI H. NO 9-119, NEAR RAM MANDIR SHAHBAZAR GULBARGA | | | |585104. | |2 |Area of Research |Pharmacology | |
3 |List of Experiments Carried out |INFLUENCE OF 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 |cENTRAL ANIMAL HOUSE M.
R. MEDICAL College, SEDAM road gulbarga-585 105 | | |Institution/import/others) |kARNATAKA | |7 |No. of Animals to be sacrificed |No sacrifice | |8 |Method of euthanasia and disposal |The Euthansia 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) | INFLUENCE OF 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 |INFLUENCE OF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF | | | |SELECTED PHENOTHIAZINES IN RATS. | |2 |Chief Investigator : | | | |Name : |Dr. K. PRASAD, | | | |Professor | | |Designation : |Dept. Of Pharmacology | | | |H. K. E. S’s College of Pharmacy, Sedam Road GULBARGA.
585 105 kARNATAKA | | |Dept/Div/Lab : | | | | |08472-221392 (o) | | | |08472-241972 (R) | | |Telephone number : |9844368785 | |3 |List of names of all individuals authorized to conduct|Dr. K. Prasad. | | |procedures under this proposal. |Dr. Nitin Mahurkar | | | |Santosh Ramesh Awasti | |4 |Funding source |H. K. E. S’s. college of pharmacy, | | | |GULBARGA-585 105. | |5 |Duration of the project : | | | |Number of months : |10 months | | |Date of initiation : |01. 06. 2010 | | |Date of completion : |31. 03. 2011 | |6 |If day by which approval is needed is less than six | | | |weeks from date of submission.
Justification for the |Doesn’t arise. | | |same. | | |7 |Study objectives (the aims of study and why they are |INFLUENCE OF SELECTIVE ANTIOXIDANTS ON EXTRAPYRAMIDAL SYMPTOMS OF | | |important) to be explained non-technical terms as for |SELECTED PHENOTHIAZINES IN RATS. | | |as possible. | | |8 |Animals required | | | |Species |Albino Rats | | |Age/weight/size |Adult Rats – 180-220 gm | | | |Either sex | | |Gender |Rats 60 | | | | | | |Number to be used |40 Days. | | | | | | |Number of days each animal will be housed | | | | | | |9 |Rationale for animals usage : | | | |Why is animal necessary for these studies?
|To find out a new safe and effective drug therapy to treat | | | |extrapyramidal disorders. | | | | | | | | | | |Why are the particular species selected? |These species are suitable for screening extrapyramidal symptoms as they| | | |possess the complications that are comparable to human beings. | | | | | | | |To meet minimal statistical requirements | | |Why is the estimated number of animals essential? | | | | | | | |Similar experiments conducted in the past. If so, |- No. – | | |number animals used and results obtained in brief. | | | | | | | |If yes, why new experiment I required?
|- No. – | | | | | | |Have similar experiment (s) been made by any other |- No. – | | |organization/agency? If so, their result in your | | | |knowledge. | | |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 |- No. – | | |transport is envisaged. | | |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 Dr. K. PRASAD, M. PHARM. PhD, Professor, Professor, H. K. E. S’s College of Pharmacy, Sedam Road, GULBARGA. karnataka- 585 105 |name |signature | |Dr. K. PRASAD, Professor, HOD (Guide) | | |
Santosh Ramesh Awasti -Student | | | | | . (For iaec/cpcsea Usage) Proposal Number: HKE COP/IAEC/19/2009-10 Date first received: 25/11/2009 Date received after modification (if any): Approval Date: 1/12/2009 Expiry Date: 31/3/2011 Name of IAEC/CPCSEA Chair person: Dr. S. Appala Raju, Principal, HKE’S COP, Gulbarga-585105 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. HKE COP/IAEC/19/2009-10.