“ANTIDIABETIC EFFECT OF SAPONINS AND ALKALOIDS OF MOMORDICA CYMBALARIA”

Synopsis for registration of M.pharm Dissertation

Submitted to

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE

KARNATAKA

In partial fulfillment

of the requirement for the Degree of

Master of pharmacy In Pharmacology

Under the Guidance of

Dr. J. Anbu

Proffesor

BY

Vijay H Solanki

I M. PHARM.

Department of Pharmacology

Karnataka College of Pharmacy, Bangalore-64

2009-10

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the candidate and address / Mr. Vijay H Solanki
S/o; Harjibhai D. Solanki, #359, Kubharwada,
Vadistreat, Vanakbara-Diu
Daman And Diu (U.T), Gujarat-362570
2. / Name of the Institution / Karnataka College Of Pharmacy
Thirumena Halli.
Hedge Nagar Main Road,
Jakkur Post,
Yalahanka Hobli,
Bangalore-560064
3. /

Course of study and subject

/ M.Pharm-Pharmacology
4. /

Date of the admission

/ 30-June-2009
5. /

Title of the topic: -

“Antidiabetic Effect of Saponins and Alkaloids of Momardica Cymbalaria.”
6.0 /

Brief resume of the intended work

7.0
8.0 / 6.1 - Need for the study:
Diabetes mellitus is a chronic metabolic disorder affecting approximately 5% of the world’s population. It is characterized by deregulation in carbohydrate, protein and fat metabolisms caused by the complete or relative insufficiency of insulin secretion and/or insulin action.
According to WHO projections, the diabetic population is likely to increase to 300 millions or more by the year 2025. Currently available therapies for diabetes include insulin and various oral antidiabetic agents such as sulfonylureas, bigunides, α-glucosidase inhibitors, glinides, which are used as monotherapy or in combination to achieve better glycaemic regulation. Many of these oral antidiabetic agents have a number of serious adverse effects. Thus, the management of diabetes without any side effects is still a challenge. [1]
Plants have always been an exemplary source of drugs and many of the currently available drugs have been derived directly or indirectly from them. The ethno-botanical information reports about 800 plants that may possess anti-diabetic activity when being assessed using the presently available experimental techniques [2]
The antidiabetic effect of saponins and alkaloids extract of Momordica Cymbalaria has not been investigated scientifically. Therefore, the present study is planned to investigate the antidiabetic effect of saponins and alkaloids extract of Momordica Cymbalaria
6.2-REVIEW OF LITERATURE:
Momordica cymbalaria, Fenzl. (Family: Cucurbitaceae) is a tuberous perenninal with very slender scandent, branched, striate, pubescent or subglabrous stem, the whole plant is acrid, with ovoid tuberous roots 3
The roots of the plants are used for menstrual irregularits, antifertility, antivulantary and abortificient activites4,5. and also ethanolic root extracts of momordica cymbalaria have antiimplantation activity6. Fruit powder and extract of Momordica cymbalaria (MC) are reported to have antidiabetic and antihyperlipidemic activies in experimental diabetic models7,8,9.
Conventional antidiabetic agents can affect several pathways of glucose metabolism such as insulin secretion, glucose uptake by target organs as well as nutrient absorption. Recently, incretins and transcription factors such as peroxisome proliferator-activated receptors—PPAR are targets of modern therapy. Insulin receptor, glucose transporters, however, has not been yet the focus of antidiabetic therapy. [3]
6.3-OBJECTIVE OF THE STUDY:
Objectives:
1. To Isolate Saponins and Alkaloids extract of roots of Momardica Cymbalaria.
2. To perform the acute toxicity study of Saponins and Alkaloids extract of roots of Momardica Cymbalaria
4. To study the antidiabetic effect of Saponins and Alkaloids extract of roots of Momardica Cymbalaria by estimating the following parameters.
·  Serum Blood glucose
·  Serum Total cholesterol
·  Serum HDL cholesterol
·  Liver glycogen
·  Enzyme activity
MATERIALS AND METHODS:
7.1 Sources of data:-
The sources of data will be obtained from:-
I.  Laboratory based studies.
II.  National and international journals.
III.  Internet.
7.2 Method of collection of data (including sampling procedure if any):-
Field and laboratory studies:
Field work:
The roots of Momardica Cymbalaria will be collected and it will be authenticated.
Laboratory work: -
Animals
Male albino wistar Rats (150-250 g) will be selected for all the experiments. They will be housed in cages on a 12-h light: 12-h dark cycle and will be allowed free access to laboratory diet.
Solvents used
Pet-ether (60-80), chloroform, ethyl acetate, methanol and water
Chemicals and reagents used
Glyburide, Glucose, Ethanol, Perchloric acid, Anthrone reagent, Sucrose/EDTA buffer, Glucose-6-phosphate, Imidazole buffer, TCA: ascorbate buffer, Ammonium molybdate, Sodium citrate, Tris-magnesium chloride buffer, Glucose-6-phosphate dehydrogenase,
Data analysis
Statistical analysis of the data will be evaluated by analysis of variance (ANOVA) followed by Dunnett’s test.
METHODOLOGY:-
1.  Preparation of the Saponins and Alkaloids extract of roots of Momardica Cymbalaria.

SAPONINS EXTRACTION ;

Dried root powder was extracted through successive solvent extraction in Soxhlet apparatus using the solvents Pet-ether (60-80), chloroform, ethyl acetate, methanol and water. The extracts were concentrated in rotary vacuum evaporator at temperature not exceeding 45oC and dried under high vacuum. The dried extracts from ethyl acetate and methanol will be combined and fractionated between butanol and water. The extract will be processed by column chromatography separately to isolate saponin [4]
ALKALODS EXTRACTION ;
Dried powdered roots of MC is extracted with cold methanol (CH3OH) Methanol extraction was continued until the plant material gave a negative test for alkaloids (Mayer’s Test). After filtration, the solvent was removed under reduced pressure at 40oC,
The crude alkaloid mixture was then separated from neutral and acidic materials, and water
solubles, by initial extraction with aqueous acetic acid (CH3CO2H) followed by dichloromethane
extraction and then basification of the aqueous solution Crude alkaloidal mixture is purified by fractional crystillisation and chromatographic separation [5]
2.  Pharmacological study design:-
I. Acute toxicity study of Saponins and Alkaloids extract of roots of Momardica Cymbalaria
·  Acute toxicity study will be conducted according to OPPT guidelines.
II. Influence of Saponins and Alkaloids extract of roots of Momardica Cymbalaria on streptozotocin-induced diabetic rats [6]
·  Healthy adult Wistar male rats will be divided into five groups I, II, III, IV and V of 6 rats each.
·  Induction of experimental diabetes: [7]
Rats of groups II, III, IV and V will be injected with a single intraperitoneal injection of 60 mg/kg of streptozotocin. Rats will be used to evaluate the antidiabetic activity when the elevated glucose level in plasma will be effective and permanent.
Group I : Normal control group receiving normal saline
Group II : Diabetic control group receiving normal saline
Group III : Diabetic group receiving insulin
Group IV : Diabetic group receiving alkaloid
Group V : Diabetic group receiving saponins
Study involves administration of vehicle, teat drugs for a period of 21 days. Serum glucose levels will be estimated on days 7, 14 and 21. Mean change in serum glucose will be calculated.
On completion of the treatment, blood sample will be collected and the following parameters for all the groups of animals will be measured. [8]
·  Lipid profile
·  HDL cholesterol
Animals will be sacrificed and liver will be dissected out and will be used for biochemical assays of the following parameters:
·  Glycogen level
·  Glucose-6-Phosphate
·  Hexokinase
7.3 - Does The Study Require Any Investigation To Be Conducted On Patients Or Animals? If So, Please Describe Briefly.
No; It is invitro study
7.4 - Has Ethical Clearance Been Obtained From Your Institution In Case Of 7.3?
Yes. Taken from Institutional Ethical Committee
LIST OF REFERENCES:-
1.  Sy GY, Cisse A, Nongonierma RB, Sarr M, Mbodj NA, Faye B. Hypoglycemic and antidiabetic activity of acetonic extract of Vernonia colorata leaves in normoglycaemic and alloxan-induced diabetic rats. J Ethnopharmacol. 2005; 98: 171-75.
2.  Firdous M, Koneri R, Sarvaraidu CH, Harish M, Shubhapriya KH. NIDDM Antidiabetic activity of Saponins of Momordica Cymbalaria in Streptozotocin-Nicotinamide NIDDM Mice. Journal of Clinical and Diagnostic Research. [serial online] 2009 April [cited: 2009 April 6]; 3: 1460-1465.
3.  Koneri R, Saraswathi CD, Balaraman R, Ajeesha EA. Antiimplantation activity of the ethanolic root extracts of momordica cymbalaria Fenzl in rats. Indian J Pharmacol. 2007; 39(2): 90-96.
4.  Sari H, Sirpa OK, Marina IH, Hannu MM, and Riitta AT. Content of the flavanols quercetin, myricetin and kaemferol in 25 edible berries. J Agri Food Chem. 1999; 47(6): 2274-79.
5.  Surya H, John B, Bremner. Initial Studies on Alkaloids from Lombok Medicinal Plants Molecules ISSN 1420-3049 © 2001 by MDPI.
6.  Badole S, Patel N, Bodhankar S, Jain B, Bhardwaj S. Antihyperglycemic activity of aqueous extract of leaves of Cocculus hirsutus (L.) Diels in alloxan-induced diabetic mice. Indian J Pharmacol. 2006; 38: 49-53.
7.  Akbarzadeh A, Norouzian D, Mehrabi MR, Jamshidi Sh, Farhangi A, Allah Verdi A, Mofidian SMA, Lame Rad B. Induction of Diabetes By Streptozotocin In Rats. Indian Journal of Clinical Biochemistry. 2007; 22: 60-64.
8.  Sharma B, Vishwanath G, Salunke R, Roy P. Effects of flavonoid-rich extract from seeds of Eugenia jambolana (L.) on carbohydrate and lipid metabolism in diabetic mice. Food Chemistry. 2008; 110: 697–705.
9 / Signature of the candidate / (VIJAY H. SOLANKI)
10 / Remarks of the Guide: The topic selected for dissertation is satisfactory. Adequate equipment and chemicals are available to carry out the project work.
11 / Name and Designation of guide: / Dr. J. ANBU
Professor
Dept of Pharmacology,
Karnataka college of pharmacy,
Bengaluru 64
Signature: / (Dr. J. ANBU)
Co-Guide: / Dr. RAJU KONERI
Head of the Department: / Dr. RAJU KONERI
Professor & Dean.
Dept of Pharmacology,
Karnataka college of pharmacy.
Bengaluru-64
Signature / (Dr. RAJU KONERI)
12 / Remarks of the Principal: All the required facilities will be provided to carry out dissertation work under the supervision of guide.
13. / Principal: / Dr. K. RAMESH,
principal
Karnataka College of Pharmacy,
Thirumenahalli,
Bangalore –64.
Signature of the Principal:
/ (Dr. K. RAMESH)

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