RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA,

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR

DISSERTATION

1.Name of the Candidate

and Address JIGNASA PATELa. Permanent Address

AVADH Block no -4, Geeta nagar -2, B/h Lal Baugh, Near Sardar Baugh,

Junagadh-362 001,

Gujarat.

b. Postal Address

Shree Devi College of Pharmacy,

Airport Road, Kenjar Village,

Malavoor Panchayat,

Mangalore-574 142

Karnataka.

2.Name of the Institute Shree Devi College of pharmacy, Airport Road, Kenjar Village,

Malavoor Panchayat,

Mangalore-574 142

Karnataka.

3.

Course of Study and Subject

Master of Pharmacy in Pharmacology

4.

Date of Admission to Course

November 2010

5.Title of the Topic: EFFECT OF MEDOHAR VATI - POLY HERBAL FORMULTION ON ANTIHYPERLIPIDEMIC AND CARDIOPROTECTIVE ACTIVITY OF ATORVASTATIN

6.

BRIEF RESUME OF THE INTENDED WORK:6.1 Need of study:

Hyperlipidemia can be defined as high level of cholesterol and high triglycerides, which are an important risk factor for the initiation and progression of atherosclerosis and ischemic heart disease1. Cardiovascular disease and the resultant complication represent the leading cause of morbidity and mortality in developed countries. Moreover, with advanced life style in developing countries, such as India, particularly in metropolitan cities hyperlipidemia and the resultant pathological condition is making increasingly important contribution to mortality statistics of such countries2.

With the advancement of recent scientific knowledge Statins, Fibrates are the best therapeutic options for Hyperlipidemia. With the boon of modern medicines they are also associated with the several side effects as a curse which demands at most medical attention; statins can cause some common side effect such as GI disturbances, myalgia and elevated liver enzyme3.

During a last decade the practice of herbalism has been witnessed in many parts of the world particularly developed countries. Herb and the herbal remedies demonstrate a significant improvement in the quality of life of a patient particularly in therapeutic challenge disease like cancer, hepatic damage, Parkinsons disease and heart diseases.Moreover such products had been used with apparent safety in traditional societies for many centuries, their introduction into western countries; where they are being combined with pharmaceutical agents possess the possibility of potential interaction between the two groups of substances. The possibility of herbdrug interactions is theoretically higher than drugdrug interactions, could be because synthetic drugs usually contain single chemical entities4,5.

Medohat vati is marketed poly herbal formulation, which claims that it is having antihyperlipidemic activity and it is also used for the treatment of CHD6. Till date there is no scientific data available against Medohat vati for antihyperlipidemic and cardioprotective activity in presence or absence of any scientific drug.

So the present study is designed to demonstrate the effect of Medohar vati on antihyperlipidemic and cardioprotective activity of atorvasatin during induced hyperlipidemia and myocardial dysfunction in different experimental models of animals. 6.2 Review of literatureMedohar vati is marketed poly herbal formulation manufactured by Swami Ramdevs Divya Pharmacy. Medohar vati significantly shows antihyperlipidemic activity and it also benefits in thyroid disorders, rheumatic arthritis and joint pain6.

It is formulated by combining different herbs and in addition to their own activity they also contribute towards antihyperlipidemic activity.

Medohar vati contains following active ingredients:- Haritaki(Terminalia chebula)/Harad(Chebulic myrobalan): Biological source: It is dried mature fruit of Terminalia chebula belonging to family Combretaceae. Chemical constituent: Chebulic acid, chebulagic acid, gallic acid.

Uses: Anti cancer, anthelmintic, astringent, purgative.

It improves digestive system and reduces cholesterol and fat7.

Bhibhitaki(Terminalia bellerica)/Baheda(Belleric myobalan): Biological source: It is dried ripe fruit of Terminalia belerica Linn, belonging to family Combretaceae. Chemical constituent: Gallic acid, chebulagic acid.

Uses: Anti oxidant, anticaries, dyspepsia, purgative.

It shows rejuvenative activity, higher amount of vitamin C and it is natural colon cleanser8.

Amalaki(Embica officinalis)/Amla(Indian gooseberry):

Biological source: It is dried as well as fresh fruits of Emblica officinalis, belonging to family Euphorbiaceae. Chemical constituent: Vitamin-C, phyllemblin, tannins.

Uses: Anti viral, in jaundice, dyspepsia, anemia, asthma, dysentery.

It corrects liver metabolism and helps body to remove fat deposition9.

Kutki(Picrorhiza kurroa): Biological source: Picrorrhiza consist of dried rhizomes of the plant Picrorrhiza kurroa, belonging to family Scorphulariaceae. Chemical constituent: Picrosides, kutkside.

Uses: Hepatotoxicity, jaundice, bitter tonic, anti bacterial.

It stimulates digestion and scrapes toxic accumulation from intestine and blood, reduces fat10.

Nishodh(Operculina turpethum): Biological source: It consists of dried roots of Ipomoea terpathum, belonging to family Convolvulaceae. Chemical constituent: Flavanoids, tannins, sterols, resins, amino acids. Uses: Hepatoprotective11, anti bacterial, in snake bite.It reduces cholesterol level12.

Vagevidanga(Embelia ribes): Biological source: It consists of dried fruits of the plant Embelia ribes, belonging to family Myrsinaceae. Chemical constituent: Embelin, vilangin.

Uses: Anthelmentic, antifertility action, anti bacterial, carminative, stimulant, dyspepsia. It clears gas from intestine13.

Shuddh guggulu(Commiphora mukul): Biological source: It is a gum-resin obtained by incision of the bark of Commiphora mukul, belonging to family Burseraceae14. Chemical constituents: Aromatic essential oil, resins, steroids. Uses: Antiseptic, uterine stimulant, diaphoretic, chronic tonsillitis, ulcerated throat. It reduces cholesterol and prevents accumulation, it regulates fat metabolism15.

Shilajeet(Asphalatum): Biological source: It is herbo mineral drug ejected out of fissures in iron rich rocks, during hot weather16.

Chemical constituents: Phospholipids, triterpenes, phenolic acids, fulvic acid, humins and humic acids, trace elements.

Uses: Antiulcerogenic, immunomodulator, antioxidant, analgesic, anti-inflammatory. It has scarping effect on fat tissue and balance overweight17.

Babool gaund(Acacia arubica): Biological source: It is dried gummy exudation obtained from the stem and branches of Acacia Arabica Wild, belonging to family Leguminosae.

Chemical constituents: Gluconic acid, galactose, arabinose. Uses: Tooth disorders, eczema, tonsillitis, diarrhoea.It helps to reduce cholesterol18.

Medohar vati acts by removing disorders of digestive system and then reduces the extra fat in the body.

Statins are most commonly used drug in treatment of Hyperlipidemia. They are competitive antagonists of HMG Co A and lower the blood serum level of LDL by inhibiting cholesterol biosynthesis and by stimulating the receptor-mediated uptake of LDL19.

Atorvastatin is long lasting inhibitor of HMG Co A reductase which has a T of about 18 to 24 hours and reduces the LDL-C 39% to 60%. Clinically it is used to reduce serum cholesterol and in severe drug- resistant dyslipidemia but it also causes some side effect such as myalgia, elevated liver enzyme, headache, GI disturbances20.

6.3 Objective of studyThe objective of the proposed study is to investigate the effect of the poly herbal formulation Medohar vati on antihyperlipidemic and cardioprotective activity of atorvastatin in different experimental models of animals. SPECIFIC OBJECTIVES: To determine the phytochemical constituents of Medohar vati.

To arrive at the therapeutic dose after acute toxicity studies following WHO guidelines. To explore the influence of Medohar vati on antihyperlipidemic and cardioprotective role of Atorvastatin using experimental models such as Triton induced hyperlipidemia and High fat diet induced obesity and hyperlipidemia in rats. To study the effect of Medohar vati on protective activities of Atorvastatin during ECG changes.

7.MATERIALS AND METHODS:7.1 Source of Data:Wistar albino rats (180 to 200 g) of either sex will be procured for this study from committee for the purpose of control and supervision of experiments on animals (CPCSEA) licensed animal house. They will be maintained under standard conditions (temperature 22 2c, relative humidity 605% and 12 h light/dark cycle). The animals will be housed in sanitized polypropylene cages containing sterile paddy husk as bedding. They will have free access to standard pellet diet and water ad libitum. The Institutional Animal Ethics Committee approved the experimental protocol. All the animals received humane care according to the criteria outlined in the Guide for the Care and Use of Laboratory Animals prepared by the National Academy of Sciences and published by the National Institute of Health. All the procedures will be performed in accordance with Institutional Animal ethics committee constituted as per the direction of the CPCSEA, under ministry of animal welfare division. Government of India, New Delhi, India.

7.2 Method of Collection of Data:

Chemicals and reagents will be procured from standard companies. Poly herbal formulation of Medohar vati will be obtained from Swami Ramdevs Divya Pharmacy and it will be subjected to different methods of evaluation of antihyperlipidemic activity as discussed in objectives.

7.3 Experimental models:1. Acute toxicity studies21:

The dose will be selected by the acute toxicity studies following limit test procedure according to OPPTS (Office of Prevention, Pesticide and Toxic Substance) guidelines. Briefly, a test dose of 2 g/kg and 5 g/kg will be given to the mice.1/10th of the maximum safe dose (either 2 g/kg or 5 g/kg) will be used as a therapeutic dose. 2. TRITON INDUCED HYPERLIPIDEMIA22 :

Experimental Design:

The animals will be randomly divided into 5 groups of 6 each. From group II to V, the rats will be injected intraperitonialy with Triton WR 1339 (400 mg /kg) and test and standard drug will be given simultaneously. Exactly after 18 hrs blood samples will be collected form rats using mild ether anesthesia. Serum will be separated by centrifugation and analyzed for various biochemical parameters. The group I will be served as normal control. The different groups will be assigned as described below.Group I : Vehicle control (1% Tween 80)

Group II : Positive control, Triton WR 1339

Group III : Atorvastatin (100mg/ kg) + Triton WR 1339

Group IV : Formulation Medohar vati + Triton WR 1339

Group V : Formulation Medohar vati + Atorvastatin + Triton WR 1339 All the drug solutions will be prepared using 1% v/v Tween 80 as emulsifying agent and given at volume of 0.2 ml/ 200g once daily.

EVALUATION:The various biochemical parameters will be evaluated are: blood glucose, phospholipids, free fatty acids, total serum lipid, total cholesterol, total proteins, triglycerides, LDL Cholesterol, HDL Cholesterol.3. HIGH FAT DIET INDUCED OBESITY AND HYPERLIPIDEMIA IN RATS3:

Experimental Design:

The Wistar rats weighing between 140-160 gm will be randomly divided into 5 groups of 6 each. From group II to V animals will be fed with high fat diet instead of normal diet. The group I will be served as normal control. The different groups will be assigned as described below.Group I : Vehicle control (1% Tween 80)

Group II : Positive control (high fat diet)

Group III : Atorvasatin (100mg/kg) + high fat diet

Group IV : Formulation Medohar vati + high fat diet

Group V : Formulation Medohar vati + Atorvastatin + high fat diet

All the drug solutions will be prepared using 1% v/v Tween 80 as emulsifying agent and given at volume of 0.2 ml/ 200g once daily.

EVALUATIONThe various parameters will be evaluated are: Body weight: Body weight will be measured initially and then at 7th, 14th,21st and 28th day of the experiment. Body temperature: Body temperature will be recorded on 28th of day afterdrug administration at 60 & 90 min with a contact time of 1 min with thehelp of a rectal telethermometer.

Biochemical parameters: On 29st day after fasting for 12 hrs the blood sample will be collected using mild ether anesthesia. Serum will be separated by centrifugation and subjected for following analysis- blood glucose, phospholipids, free fatty acids, total serum lipid, total cholesterol, total proteins, triglycerides, LDL Cholesterol, HDL Cholesterol. The liver will be isolated and evaluated for total lipid and total cholesterol.ECG Studies23:After anesthetizing the rat with a combination of ketamine hydrochloride (75mg/kg, i.p) and xylazine (8.0mg/kg, i.p), leads will be attached to the dermal layer of both the front paws and hind legs and recording will be made with the help of computerized ambulatory ECG system. Animal grouping and treatment is same as triton induced and high fat diet induced Hyperlipidemia models.

7.4 Does the study require any investigation or interventions to be conducted on patients or the human or animals? If so please describe briefly:

YES

Study requires investigation on animals. The effects of the drug will be studied on various parameters using rats and mice as experimental animals.

7.5 Has ethical clearance been obtained from your institute

Ethical Committee approval letter is enclosed.

8.List of References:1. Goldstein JL, Schrott HG, Hazzard WR, Bierman EL, Motalsky AG. Hyperlipidemia in coronary heart disease 2. J chin Invest 1973 July;52(7):1544-68.

2. Braumwald E3 editor, Heart disease: A textbook of cardiovascular medicine 5th ed. Philadelphia: W.B. Saunders co;1997.3. Parasuraman S, Kumar EP, Kumar A, Emerson SF. Antihyperlipidemic effect of Triglize poly herbal formulation. Int J Pharm Pharm Sci 2010;2(3):118.

4. Singh YN. Potential for interaction of kava and St. Johns wort with drugs. J.Ethnopharmacol 2005;100:10813.5. Izzo AA. Herbdrug interactions: an overview of the clinical evidence. Fund & Cli Pharmacol 2005;19 (2):1-16.6. htpp://www.lovenaturalremedies.com/Divya-Yog-Mandir-Trust-Swami-Ramdev/Herbal-/-ayurvedic/Other-medicines/Divya-MedoharVati-50-gm.html retrieved at 3/5/2011 at 1.05p.m.7. Kokate CK, Purohit AP, Gokhle SB. Pharmacognosy. 44th ed. 2009. p. 9.4.8. Kokate CK, Purohit AP, Gokhle SB. Pharmacognosy. 44th ed. 2009. p. 9.6.9. Pandey G, Pandey SP. Phytochemical and Toxicity study of Emblica officinalis. Int Res J Pharm 2011;2(3):270-72.10. Banerjee D, Maity B, Nag SK, Bandyopadhyay SK, Chattopadhyay S. Healing potential of Picrorhiza kurroa rhizomes against indomethacin induced gastric ulceration. BMC Complement Altern Med 2008;8:3.

11. Haran M, Gafur MA, Sadik MG, Raheman AA. Antibacterial and Cytotoxic activities of extracts and isolated compounds of Ipomoea turpathum. Pak. J. Bio. Sci. 2002;5(5):597-99.

12. Evance WC. Trease and Evance Pharmacgnosy 5th ed. 2005. p. 473.

13. Khan MI, Ahmed A, Akram M, Mohiuddin E, Khan U, Ayaz S et al. Monograph of Embelia ribes. Afr. J. Plant. Sci. 2010;4(12):503-05.

14. Ali M. Textbook of Pharmacognosy 2nd ed. 1998. p. 386-87.

15. Dwivedi A, Dixit SK. A study on Guggulu with special reference to Analgesic effect AYU 2009;30(1):5-10.

16. Kokate CK, Purohit AP, Gokhle SB. Pharmacognosy. 44th ed. 2009. p. 18.1-18.9.

17. Talbert R. A material medica monograph 2004.

18. Ali HB, Ziad A, Blunden G. Biological effect of gum Arabic. Food Chem. Toxicol. 2009;47:1-8.

19. Rang HP, Dale MM, Ritter JM, Flower RJ. Rang and Dales pharmacology. 6th ed. Churchill Livingstone 2007. p. 325-26.

20. Tripathi KD. Essential of medical pharmacology. 5th ed. New Delhi: 2003. p. 578.

21. Kokate CK, Practical Pharmacognosy. 3rd ed. New Delhi: VPBN 1991. p. 107-11

22. Kumar V, Singh P, Chander R, Mahdi F, Singh S, Singh R et al. Hypolipidemic activity of Hibiscus rosa sinensis root in rats. Indian J. Biochem. Biophys. 2009;46:507-10.23. Singh PN, Athar MS. Simplified calculation of mean QRS vector (mean electrical axis of heart) of electrocardiogram. Indian J Physiol Pharmacol 2003;47:212-16.

9.

SIGNATURE OF THE CANDIDATE:

10. REMARKS OF THE GUIDE:EFFECT OF MEDOHAR VATI - POLY HERBAL FORMULTION ON ANTIHYPERLIPIDEMIC AND CARDIOPROTECTIVE ACTIVITY OF ATORVASTATIN to be carried out by Miss Jignasa Patel of M. Pharm has been discussed and worked out under my directions and supervision as an official guide. The project work envisaged is of great importance in the field of herbal research. The work can be carried out in pharmacology laboratory of Shree Devi College of Pharmacy for which facilities are available. Hence the project is viable and is recommended for clearance and approval.

11. Name and Designation of

11.1 GuideDr. Jagadish V. KamathDept. of Pharmacology.Shree Devi college of pharmacy

Air port road, Kenjar village.

Malavoor panchayath,

Mangalore-574 142

11.2 Signature

11.3 Head of the Department

Dr. Jagadish V. KamathDept. of Pharmacology.Shree Devi college of pharmacy

Air port road, Kenjar village.

Malavoor panchayath,

Mangalore-574 142

11.4 Signature

12. 12.1 Remarks of the Principal

The Programme and the Research work that is undertaking by Miss Jignasa Patel has potential implication in the field of Pharmacology. The work can be carried in the Research Laboratories of Pharmacology Department at Shree Devi college of Pharmacy. Hence the Project is recommended and requested for clearance and approval.

12.2 Signature

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