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“COMPARISON OF ANTICANCER ACTIVITY OF TURMERIC AND MINT USING EHRLICH ASCITES CARCINOMA CELLS”




SYNOPSIS FOR

M.PHARM DISSERTATION



SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA

BY

SOMAYEH AFSAH VAKILI

I M.PHARM

DEPARTMENT OF PHARMACOLOGY

VISVESWARAPURA INSTITUTE OF

PHARMACEUTICAL SCIENCES

BANGALORE-560070

(2011-2012)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA.

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION


NAME OF THE CANDIDATE AND ADDRESS


SOMAYEH AFSAH VAKILI

#S104, SHALINI TOWER, BNASHANKARI 2nd STAGE, BANGALORE-560070



NAME OF THE INSTITUTION



VISVESWARAPURA INSTITUTE OF

PHARMACEUTICAL SCIENCES

BANGALORE-560070


COURSE OF THE STUDY

& SUBJECT


MASTER OF PHARMACY IN

PHARMACOLOGY



DATE OF ADMISSION


19/05/2010


TITLE OF THE TOPIC:

“Comparison of anticancer activity of turmeric and mint using Ehrlich Ascites Carcinoma cells”


BRIEF RESUME OF INTENDED WORK

NEED FOR STUDY
Cancer is the second largest single cause of death, claiming over six million lives each year worldwide. Depending on the stage of cancer, surgery, radiotherapy and chemotherapy are the most commonly used treatment modalities1.

In the present scenario the search for newer molecules is changing towards the natural bioactive substances owing to their fewer side effects and low cost. About 60% of currently existing anticancer drugs are derived from natural sources2.
Turmeric(Curcuma longa) and Mint(Mentha Piperita) are frequently used in our diet. Anticancer activity of these two functional foods has been proved 3,4. The present study is taken up to compare the anticancer activity of turmeric and mint in the EAC tumor bearing mice as per the standard protocol recommended by the National Cancer Institute, USA. This will enable us to suggest to increase the frequency of the more potent of the two in diet for prophylactic/curative purpose.
Review of Literature:

Herbal drugs constitute a major share of all the officially recognised systems of health in India viz. Ayurveda, Yoga, Unani, Siddha, Homeopathy and Naturopathy, except Allopathy. More than 70% of India’s 1.1 billion populations still use these non-allopathic systems of medicine. Currently, there is no separate category of herbal drugs or dietary supplements, as per the Indian Drugs Act. Significant basic and clinical research has been carried out on the medicinal plants and their formulations, with the state-of-the-art methods in a number of Institutes/Universities5.
Turmeric is dried as well as fresh rhizomes of the plant known as Curcuma longa Linn. Belonging to the family Zingiberaceae. It contains about 5% of volatile oil, resins8. In traditional Indian Ayurvedic medicine, turmeric has been used as a tonic for the digestive system and the liver; to dispel worms, strengthen the body, and dissolve gallstones; and for menstrual irregularity and arthritis. In old Hindu texts it is described as an aromatic, stimulant, and carminative (an agent that helps expel gas from the intestines and treats colic). Mixed with slaked (hydrated) lime, turmeric was a well known household remedy for sprains and swellings caused by injury7. It has been proved to posses anti-inflammatory, antiarthritic6. antioxidant, antifungal8 and anticancer activities3,6.
Mint a widely available herb with a number of medicinal properties and helps to promot good health. The oil is obtained by steam distillation of the fresh flowering tops of plants known as Mentha piperita Linn. Belonging to the family Labiatae. It contains volatile oils majorly l-menthol. Mentha oil is traditionally used as carminative, stimulant and flavouring agent6. It proved to posses antiseptic, antimicrobial9 and anticancer activities10.
Objective of the study:

The objectives of the present study are as follows :

  1. Comparison of anticancer activity of turmeric and mint, using EAC.

  2. Evaluation of antioxidant enzyme in turmeric and mint pretreated mice.


MATERIALS & METHODS
Source of data

Whole work is planned to generate data from laboratory studies i.e.; experiments are performed as described in reference, experimental studies in journals and in textbooks available with college, IISc library, RGUHS digital library (Helinet), websites www.sciencedirect.com,

www.ncbi.nlm.nih.gov/pubmed,www.google.com,www.ijp-online.com,will be used to obtain related information regarding this research protocol.

METHODS OF COLLECTION OF DATA

The data collected will be based on laboratory animal experimentation
1. Preparation of drug and mode of administration:

The drugs Mentha piperita and Curcuma longa will be obtained locally and authenticated. The crude drug will be shade dried, powdered and extracted using distilled water.

2.Animals:

Swiss albino mice either sex of 6-8 weeks old (25±5 g body weight) will be used in these study. The animal experiments will be performed according to the rules and regulations of the Institutional Animal Ethics Committee1,11.
3.Tumor cells

The Ehrlich’s Ascites Carcinoma (EAC), will be obtained from, Amala Cancer Research Center, Thrissur, Kerala. They will be maintained by weekly intraperitoneal inoculation of 106 cells/mouse.

4.Anticancer effects in the Ehrlich Ascites Carcinoma bearing mice[1]

The preinoculated (1x106 cells/mice, i.p.) will be maintained for 15 days12,13. After two week of inoculation, the peritoneal fluid will be aspirated and diluted to get 1x106 EAC cell / ml in a phosphate buffer. Mice will be reinoculated with 1x106 EAC cell / mice 12 and divided into 5 groups consisting of 8 animals in each group.


Group

No. of Animals

Drug

Treatment


Group 1

8

Normal mice

-----

Group 2

8

EAC mice+ distilled water

1 ml

Group 3

8

EAC mice + cisplatin

3.5mg/kg b.wt14.

Group 4

8

EAC mice + turmeric

500mg/kg b.wt 15.

Group 5

8

EAC mice + mint

500mg/kg b.wt16.













b. wt = body weight

Group 1-5 will be treated with vehicle & drug samples orally 24 hrs after the inoculation of EAC and continued for 9 days and the observation will continue until the termination of the study.The dose of Cisplatin, turmeric and mint are selected based on literature.

TUMOR GROWTH RESPONSE1

At termination, surviving animals of EAC tumor bearing mice will be counted and the average survival time (MST)17 and the % increase in life span (% ILS) will be Calculated by following formula.

Median survival time (MST) = (Day of 1st death + Day of last death) / 2

Percentage increase in life span = [ (MST of treated group/ MST of control group) – 1] x 100
Body weight analysis: During the same time of study, weekly body weight analysis will be recorded.

% Increase in weight = [(animal weight on respective day/animal weigh on day 0) – 1] x 100
ESTIMATION OF HEMATOLOGICAL PARAMETERS1

On the last day of treatment, blood will be drawn from each mouse in the conventional way and the white blood cell count (WBC), red blood cell count (RBC), hemoglobin, differential count will be determined using Haemocytometer and Sahli’s haemometer.
ESTIMATION OF ANTIOXIDANT ENZYME18.

Lipid peroxidase , glutathione, super oxide dismutase and catalase levels will be measured in tumor cells collected from untreated mice and 1 h after treatment of tumor-transplanted mice with last dose in each group by using standard procedures.
STATISTICAL ANALYSIS:-

The data obtained from experimentation will be subjected to one way Analysis Of Variance (ANOVA) with suitable post-hoc test.




LIST OF REFERENCES

  1. Durairaj AK, Vaiyapuri TS, Mazumder UP, and Gupta M. Antineoplastic and antioxidant activities of Oxystelma esculentum on Swiss albino mice bearing Ehrlich’s ascites carcinoma. Pharmaceutical Biology 2009;47:195–202

  2. Newman DJ, Cragg GM, Snader KM. Natural products as a source of new drugs over the period of 1981-2002. J Nat Prod 2003;66:1022-37

  3. Kuttan R, et al. Potential anticancer activity of turmeric (Curcuma longa). Cancer Lett 1985;29:197-202

  4. Pamela LC. Prevention and Therapy of Cancer by Dietary Monoterpenes. J of Nutrition. 1999;129:775-8

  5. Ashok DB, Vaidya, Thomas PA, Devasagayam. Current Status of Herbal Drugs in India: An Overview. Clin Biochem Nutr 2007;41:1–11

  6. Kokate CK, Purohit AP and Gokhale SB. Pharmacognosy,13th ed. Nirali Prakashan; 1999. P. 273-5 and 430-1

  7. Ammon HP, Wahl MA. Pharmacology of Curcuma longa. Planta Med 1991;57:1-7

  8. Jayaprakasha GK, Jena BS, Negi PS and Sakariah kk. Evaluation of antioxidant activities and antimutagenicity of turmeric oil: A Byproduct from Curcumin Production. Z.Naturforsch 2002;57:828-35

  9. Sivropoulou A, Kokkini S, Lanara T and Arsenakis M. Antimicrobial activity of mint essential oils. J Agri Food Chem 1995;43:2384-4

  10. Cadenas E and Packer L. Spices as potent antioxidant with therapeutic potential. In Handbook of antioxidants. CRC press. USA (LA) 2001. pp.276-9

  11. Nandi P, Talukderl G, Sharma A. Dietary chemoprevention of clastogenic effects of 3,4- benzo(a)pyrene by Emblica officinalis Gaertn Fruit extract. Br J Cancer 1997;76:1279-83

  12. Gothoskar SV, Ranadive KJ. Anticancer screening of SAN-AB: An extract of marking nut semicarpus anacardium. Indian J Exp Biol 1971;9:372-5

  13. Babu, Kuttan TD, Jose P. Cytotoxic and anti-tumour properties of certain taxa of umbelliferae with special reference to Centella asiatica. Urban J. Ethno. Pharmacol 1995;48:53

  14. Satisha MP, Revankar VK and Pai KSR. Synthesis, structure, electrochemistry, and spectral characterization of bis-isatin thiocarbohydrazone metal complexes and their antitumor activity against ehrlich. Ascites carcinoma in swiss albinomice. Metal based drugs 2008;10:1155-65

  15. John TP et al, Mechanisms of anticarcinogenic properties of curcumin: the effect of curcumin on glutathione linked detoxication enzymes in rat liver. The Int J of Biochemistry and Cell Biology 1998;30:445-6

  16. Ibrahim AM et al. antisecretagogue antiulcer and cytoprotective effects of peppermint mentha piperita L. In laboratory animals. J of med sci 2006;6:930-6

  17. Majumdar UK, Gupta M, Maiti S. Antitumour activity of Hygrophila spinosa on Ehrlich ascites carcinoma and sarcoma-180 induced mice. Indian J Exp Biol 1970;35:473-76

  18. Sushma MB et al. Enhancement of radiation-induced oxidative stress and cytotoxicity in tumor cells by ellagic acid. Clinica Chimica Acta 2005;359:89–100

  19. Jagetia GC, Baliga MS, Malagi KJ, Sethukumar Kamath M. The evaluation of the radioprotective effect of Triphala (an ayurvedic rejuvenating drug) in the mice exposed to gamma-radiation. Phytomedicine 2002;9:99-108

  20. Jain. Ethnobotany and research on medicinal plants in India. Ciba Found Symp 1994;185:153-64

  21. Rachana G, Dietary turmeric modulates DMBA-induced p21ras, MAP kinases and AP-1/NF-κB pathway to alter cellular responses during hamster buccal pouch carcinogenesis. Toxicology and Applied Pharmacology 2008; 232:428–39

  22. Ghosh MN, Fundamentals of Experimental Pharmacology, third edition. Hilton and company kolkotta;2005:192




9.0

SIGNATURE OF THE CANDIDATE




10.

REMARKS OF THE GUIDE



11.


NAME AND DESIGNATION
11.1 GUIDE



Dr. MEERA SUMANTH

H.O.D

DEPT. OF PHARMACOLOGY,

VIPS, BANGALORE-560070.



11.2 SIGNATURE






11.3 CO-GUIDE (IF ANY)






    1. SIGNATURE

.




    1. HEAD OF THE

DEPARTMENT


Dr. MEERA SUMANTH

H.O.D

DEPT. OF PHARMACOLOGY,

VIPS, BANGALORE-560070.



11.6 SIGNATURE





12.

12.1 REMARKS OF THE

PRINCIPAL









12.2 SIGNATURE

Prof. Dr. D. H. HARISH KUMAR

PRINCIPAL,

VIPS, BANGALORE-560070.





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