Rajiv Gandhi University of Health Sciences s26

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE, KARNATAKA

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. Name of the candidate and MR. ABHINIT KUMAR address DEPT.OF PHARMACOGNOSY AND PHYTOCHEMISTRY, K.L.E.SOCIETY’S COLLEGE OF PHARMACY, J.N.M.C.CAMPUS, NEHRU NAGAR, BELGAUM-10.

2. Name of the Institution K.L.E.SOCIETY’S COLLEGE OF PHARMACY, BELGAUM-10

3. Course of study and MASTER OF PHARMACY IN PHARMACOGNOSY subject AND PHYTOCHEMISTRY

4. Date of admission to course JUNE-2007

5. Title of the topic “PHYTOCHEMICAL INVESTIGATION AND ANTI-CANCER ACTIVITY OF TERMINALIA CHEBULA Retz.”

6. BRIEF RESUME OF THE INTENDED WORK: INTRODUCTION:

Cancer is a Latin word meaning a crab. The Greek word for a crab is “Karkinos” and the Sanskrit word is “karkara”. Cancer means any malignant tumour, irrespective of its nature. A malignant tumour, alike the crab, has a fat main body with extensions, like the craps feet, which invade the surrounding tissues. There is a difference between cancer and carcinoma: a carcinoma is a cancer but specifically means an epithelial malignant tumour, while the term cancer, as mention above, means any malignant tumour – carcinomas, sarcomas, malignant, teratomas, embryonic malignant tumours, leukaemias and others1. The National Cancer Institute (NCI) recently announced that the death rate from cancer in the united states fall by roughly 3% between 1991-1995, the sustain decline since national record- keeping begin in 19930’s. most of the over all drop is due to decline in lungs, colorectal and prostate cancer death’s in men, and breast, colorectal and gynaecologic cancer death’s in woman. Lung cancer motility falls by 6.7% in men in the 5 year period, while rising by 6.4% in women1. An extremely promising strategy for cancer prevention today is chemoprevention, which is defined as the use of synthetic or natural agents (alone or combination) to block the development of cancer in humans. Plants, vegetables and herbs used in the folk and traditional medicine have been accepted currently as one of the main source of cancer chemoprevention drug discovery and development2. The agents discovered in the first two decades of cancer chemotherapy (1950-1970) largely interacted with DNA or its precursors, inhibiting the synthesis of new genetic material or causing irreparable damage to DNA itself. In recent years, the discovery of new agents has extended from the more conventional natural products such as Paclitaxel and semi synthetic etoposide, both of which target the proliferative process, to entirely new field of investigation that represent the harvest of new knowledge about cancer biology. The first successful application of this knowledge include diverse drug. One agent, interleukin-2, regulates the proliferation of tumor-killing T-lymphocytes and so-called natural killer cell; this agent has proven able to induce remission in fraction of patients with malignant melanoma and renal cell carcinoma, diseases unresponsive to conventional drugs3. 6.1 NEED FOR THE STUDY: Medicinal plants are natural resources yielding valuable herbal products, which are offen use in the treatment of various aliments crud material of these herbs are used in the ayurvedic preparations. Medicinal plants have been used in the treatment of various degenerative diseases such as cancer, inflammation and liver diseases. Large number of plant have been reported to possess anti-tumour activity in laboratory animals 4. The survey of literature reveals that the plant Terminalia chebula Retz. belongs to the family Combretaceae popularly known as harra (Hindi) has a widespread occurrence in North India, also occur in Bihar, West Bengal, Assam, Central India and South India5 . It is used in traditional medicine for the treatment of various ailments. The fruit of the plant is an astringent and aperient used in the treatment of dysentery and diarrhea. The ripe fruit is purgative, tonic, carminative, enrich the blood, good in ophthalmia, diseases of spleen, piles and use in paralysis6. The various chemical constituents of Terminalia chebula present in fruit are Anthraquinone glycoside, chebulinic acid, tannic acid, terchebin, vitamine C. fruit kernels contain arachidic, behenic, linoleic, oleic, plamitic and stearic acid5. Plant derived natural products such as flavonoids, terpenoids and steroids etc have received considerable attention in recent year due to their diverse pharmacological properties including antioxidant and anti-tumor activity2. However, the anti-tumor activity of methanol extract of Terminalia chebula fruit was evaluated against various cancer cell lines7, has been scientifically investigated. Hence, the present study is undertaken for the phytochemical investigation of aqueous and petroleum ether extract of fruit of Terminalia chebula on different cancer cell lines to evaluate its traditionally claimed anti-tumor activity6. 6.2 REVIEW OF LITERATURE: Terminalia chebula Retz. (Family- Combretaceae) Vernacular Name : Sanskrit - Haritaki Bengali - Haritaki Hindi - Harra Tamil - Kadukkai5 Terminalia chebula Retz. Is an astringent, a large tree; young branchlets, leaf-buds, and leaves with long, soft, shining, rust-coloured, sometimes silvery hairs. Leaves mostly subopposite, distant, ovate or oblong-ovate, 8-20 cm long, deciduous in the cold season. Flower dull-white or yellowish, with a strong offensive smell, brone in spikes from the upper axils and in small terminal panicles. Fruits obovoid or ellipsoidal from a broad base, glabrous, more or less 5-ribbed when dry5. The fruit of this plant are reported to be medicinally important in the traditional system of medicine and are used extensively for used in the treatment of dysentery and diarrhea. The ripe fruit is purgative, tonic, carminative, enrich the blood, good in ophthalmia, diseases of spleen, piles and use in paralysis5,8. The various chemical constituents of Terminalia chebula present in fruit are Anthraquinone glycoside, chebulinic acid, tannic acid, terchebin, vitamine C. fruit kernels contain arachidic, behenic, linoleic, oleic, plamitic and stearic acid5. Review of literature revels that Terminalia chebula extract show antibacterial activity9, anticaries agent10, antioxidant11, wound healing activity12, purgative action13, hepatoprotective14, anti-diabetic activity15, immunomodulatory activities16. There are many plant exracts which show potent anti-cancer effect like Oronylum indicum17, Brucea antidysentrica18, Taxus brevifolia, Taxus baccata19, Toona ciliate20, Carica papaya, Coleus tuberosus, Mangifera indica, Eugenia Polyantha21, Scutellaria barbata22.

6.3 OBJECTIVE OF STUDY: The objectives of the present study are  Collection and identification of plant.  Collection and processing of stem bark of Bauhinia racemosa.  Successive solvents extraction of the stem bark of Bauhinia racemosa.  Pharmacological screening of stem bark of Bauhinia racemosa for anti-cancer activity.  Separation of major chemical constituents from potent extract by Chromatography.  Characterization of separated chemical constituent from potent extract spectroscopic methods.

7. MATERIALS AND METHODS:

7.1 SOURCE OF DATA The source of data for this study is based on cancer cell lines; also the data obtained from the literature will be the source of data. 7.2 METHOD OF COLLECTION OF DATA (Including sampling procedure if any) The fruit of the Terminalia chebula selected for the study will be collected from Jammu region and were authenticated. STANDARDIZATION OF TERMINALIA CHEBULA FRUIT:  Botanical Evaluation: Colour, Odour, Taste, Size, Shape and Texture  Physical Evaluation: Ash value, Extractive value and Loss of drying23.  Phytochemical Evaluation: The fruit of Terminalia chebula will be collected, shade dried and powdered. The powdered material will be subjected to aqueous and petroleum ether extraction.  Phytochemical Investigation: The extracts will be subjected for preliminary phytochemical investigation to identify different phytoconstituents24. Then subjected for chromatographic techniques for then separation of phytoconstituents. Later for spectroscopic methods for the characterization of individual phytoconstituents.  Pharmacological Evaluation: Anti-cancer activity was done on different cancer cell lines25,26. Cytotoxicity testing is based on mammalian cell lines being grown under conditions where they are actively growing and undergoing mitotic division. Cells are cultured in a microtitre well plate and the rate of multiplication and growth is measured indirectly by formation of a colour, the intensity of which is directly proportional to the number of cells present. Number of experiments can be used and the most basic is to compare the rate of proliferation of a cancer cell line in the presence and absence of the test substance, usually after a specified time. Ideally several different cancer cell lines can be used so that selectivity can be assessed. This gives an indication of potential usefulness in a clinical setting. The major technique for testing cytotoxicity is sulphorhodamine B (SRB) assay. This relies on the uptake of the negatively charged pink aminoxanthine dye, sulphorhodamine B (SRB) by basic amino acids in the cells. The greater the number of cells, the greater amount of dye is taken up and, after fixing, when the cells are lysed, the released dye will give a more intense colour and greater absorbance. The SRB assay is sensitive, simple, reproducible and more rapid than the formazan-based assays and gives better linearity.

7.3 Does the study require any investigation or invention to be conducted on patients or other humans or animals? If so please mention briefly.  Not Applicable 7.4 Has ethical clearance been obtained from yours institution in case of 7.3  Not Applicable

8. REFERENCES: 1. Deodhare SG. General Pathology and Pathology of System. 6th ed. Mumbai: Popular Prakashan; 2002 : 1566-1567. 2. Gupta M, Mazumder UK, Sambath RK. Antitumor effect of Bauhinia racemosa against Ehrlich ascites carcinoma with reference to lipid peroxidation and antioxidant system in Swiss albino mice, Acta Pharmacol Sin. 2004;25(8): 1070–1076. 3. Harman JG, Limbird LE. Goodman and Gilman’s The Pharmacological Basis of therapeutics.10th ed. New york: McGraw-Hill medical publishing division; 2001: 1381. 4. Kumar RS, Gupta M. Effect of methanolic extract of Caesalpinia bonducella and Bauhinia racemosa on hematology and hepatorenal function in mice. Journal of Toxicological Sciences 2005;30(4): 265-274. 5. Chatterjee A, Pakrashi SC. The treatise on Indian medicinal plants. New delhi : Publication and information directorate;1992;Vol-2: 21-22. 6. Kirtikar KR, Basu BD. Indian medicinal plants. 2nd ed. Dehradun:International book distributors;1999; Vol-2: 894. 7. Saleem A, Husheem M. Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula retz. Fruit. Journal of Ethnopharmacology;81;2002:327-336. 8. Wealth of India, Raw Materials, 1952. Council of Scientific and Industrial Research, Publication and Information Directorate, New Delhi, 54–55. 9. Malekzadeh F, Ehsanifar H. Antibacterial activity of black myrobalan (Terminalia chebula) against Helicobacter pylori. Journal of Ethnopharmacology;81;2002:327-336. 10. Carounanidy U, Satyanarayanan R, Velmurugan A. Use of an aqueous extract of Terminalia chebula as an anticaries agent: A clinical study. Indian J Dent Res. 2007; 18(4):152-156. 11. Lee HS, Won NH, Kim KH, Lee H, Jun W, Lee KW. Antioxidant effects of aqueous extract of Terminalia chebula in vivo and in vitro. Biol Pharm Bull. 2005; 28(9):1639-1644.

12. Suguna L, Singh S, Sivakumar P, Sampath P, Chandrakasan G. Influence of Terminalia chebula on dermal wound healing in rats. Phytother Res. 2002; 16(3):227-231. 13. Miglani BD, Sen P, Sanyal RK. Purgative action of an oil obtained from Terminalia chebula. Indian J Med Res. 1971; 59(2):281-283. 14. Tasduq SA, Singh K, Satti NK, Suri KA, Gupta DK. Terminalia chebula (fruit) prevents liver toxicity caused by sub-chronic administration of rifampicin, isoniazid and pyrazinamide in combination. Human & Experimental Toxicology 2006; 25(3): 111-118. 15. Kumar S, Periasamy G. Anti-Diabetic Activity of Fruits of Terminalia chebula on Streptozotocin Induced Diabetic Rats. Journal of health science; 52(3): 283- 291. 16. Shivaprasad HN, Mohan S. Preliminary Immunomodulatory Activities of the Aqueous Extract of Terminalia chebula. Pharmaceutical Biology 2006; 44(1): 32 – 34. 17. Mao A A. Oroxylum indicum Vent.- A potential anticancer medicinal plant. Indian Journal of Traditional Knowledge 2002; 1(1):17-21. 18. Cuendet M, Pezzuto JM, Antitumor activity of bruceantin: An old drug with new promise. Journal of Natural Products 2004;67(2): 269-272. 19. Erdemoglu N, Sener B. Antitumor effects of the taxane class compounds. Journal of Faculty of Pharmacy, Ankara University 2000;29(1):77-90. 20. Chowdhury R, Hasan CM, Rashid MA. Bioactivity from Toona ciliata stem bark. Pharmaceutical biology 2003;41(4): 281-283. 21. Ali AM, Mooi LY, Yih KY. Anti-tumor promoting activity of some Malaysian traditional vegetable (ulam) extracts by immunoblotting analysis of Raji cells. Natural Product Sciences 2000;6(3):147-150. 22. Chan JYW, Tang PMK. Pehophorbide a, major antitumor component purified from Scutellaria barbata, induces apoptosis inhuman hepato cellular carcinoma cells. Planta Medica 2006;72(1):28-33. 23. Khandelwal KR. Practical Pharmacognosy Techniques and Experiment. 4th ed. New Delhi: Nirali Prakashan; 1994: 107-111. 24. Kokate CK. Practical Pharmacognosy. 4th ed. New Delhi: Vallabh Prakashan;1994: 107-111. 25. Monk A. Feasibility high flux anticancer screen utilizing diverse panel of human tumor cell lines in culture. J Natl Cancer Inst 1991;83: 757-766. 26. Skehan P, Storeng RS. New colorimetric cytotoxicity assay for anti-cancer drug creening. J Nat Cancer Inst 1990;82;: 1107–1112.

9. Signature of Candidate 10. Remark of the guide The above information and literature has been extensively investigated, verified and was found to be correct. The present study will be carried out under my supervision and guidance.

11. Name and Designation of Dr. SUNIL S. JALALPURE M.Pharm, Ph.D. (in block letters) Associate Professor, 11.1 Guide Department of Pharmacognosy & Phytochemistry, K.L.E.Society’s College of Pharmacy, Belgaum.

11.2 Signature

11.3 Co-guide Dr. K.A.SURI MSc.,Ph.D. Deputy Director , NPC Division & Chairman Instrumentation Division, Indian Institute of Integrative Medicine(RRL), Canal Road, Jammu Tawi-180001

11.4 Signature

11.5 Head of the Department Dr. SUNIL S. JALALPURE M.Pharm, Ph.D. Associate Professor & Head Department of Pharmacognosy & Phytochemistry, K.L.E.Society’s College of Pharmacy, Belgaum.

11.6 Signature

12. 12.1 Remarks of the The above mentioned information is correct and I Chairman & Principal recommend the same for approval. 12.2 Signature

Dr. F.V.MANVI M.Pharm.Ph.D. PRINCIPAL, K.L.E.Scociety’s College of Pharmacy, Belgaum-10