“CNS DEPRESSANT AND ANTIEPILEPTIC ACTIVITY OF AMORPHOPHALLUS PAEONIIFOLIUS”

M.Pharm Dissertation Protocol Submitted to

Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore– 560041

By

Mr. Vipin PankajB.Pharm

Under the Guidance of

Dr. Preeti KulkarniM.Pharm, Ph.D

Department of Pharmacology

S.E.T’s COLLEGE OF PHARMACY

S. R. Nagar, Dharwad–580002

2011-2012

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE –II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1. / NAME OF THE CANDIDATE AND ADDRESS / VIPIN PANKAJ DEPT. OF PHARMACOLOGY, SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD-580002
2. / NAME OF THE INSTITUTION / SET’s COLLEGE OF PHARMACY, S. R. NAGAR, DHARWAD-580002
3. / COURSE OF STUDY AND SUBJECT / MASTER OF PHARMACY IN PHARMACOLOGY
4. / DATE OF ADMISSION TO THE COURSE / 11-06-2011
5. / TITLE OF THE TOPIC:
“CNS DEPRESSANT AND ANTIEPILEPTIC ACTIVITY OF AMORPHOPHALLUS PAEONIIFOLIUS ”
6.0
7.0
8.0 / BRIEF RESUME OF THE INTENDED WORK
6.1Need for the study:
Depression is the most prevalent mental disorder and it is recognized to be symptomatically, psychologically and biologically heterogeneous. The disorder is characterized by apathy, loss of energy, retardation of thinking and activity, as well as profound feelings of gloominess, despair and suicidal ideation1. According to WHO approximately 450 million people suffer from a mental or behavioural disorder. This amounts to 12.3% of the global burden of disease, and will rise to 15% by 20202.
Epilepsy is a group of disorders of CNS characterized by paroxysmal cerebral dysrhythmia, manifesting as brief episodes (seizures) of loss or disturbance of consciousness, with or without characteristic body movements (convulsions), sensory or psychiatric phenomena3. Epileptic seizure is caused by non-physiological high frequency discharge of impulses by a group of neurons in the brain. Around 30,000 people develop epilepsy every year and the condition will affect about one person in 20 at some time during their lives4.
In modern system of medicine phenobarbitone, phenytoin,diazepam are widelyused to control the seizure attacks which act bydifferent mechanisms5. The use of these drugs is mostly accompanied by transient gastrointestinal symptoms, including anorexia, nausea and vomiting6. Use of synthetic antidepressant drugs like Trimipramine, fluoxetine are also reported to increase the risk of development of type 2 diabetes7.Hence it is worthwhile to explore the utility of traditional medicines in the treatment of nervous disorders.
Indian system of medicine has suggested medicinal plants and herbal formulation like Amorphophallus Paeoniifolius, which acts on CNS and helps to control depression and epilepsy8. Therefore it deserves further study on CNS depressant and antiepileptic activity on experimental animals.
6.2Review of Literature:
Amorphophallus Paeoniifolius (Family: Araceae). It is an herbaceous plant with underground hemispherical depressed dark brown corm with 60-90cm in length and 6cm width. It is cultivated in Karnataka, West Bengal, Maharashtra and in plains of North India. Leaf solitary, large, Spathe bell-shaped, greenish pink inside, purplish outside. Flower-unisexual, berried-clustered, ovoid-red, Fruits-ovoid 2-3 seeded with red color.
Corm of the plant contains lupeol, β-sitosterol, stigmasterol, betulinicacid, quercetine, kaempferol, triacontane, tannins, β-sitosterol palmitate, galactose, glucose, rhamnose and sucrose. Roots are used in opthalmia, boils and as emmenogogue. Fermented juice of petiole is used to cure diarrhea. Seeds are used in rheumatism8.
The plant has been reported to have anti-inflammatory9, antidiarrhoeal10, gastroprotective11 and anthelmintic12activity.To our best knowledge no scientific data regarding the activity of Amorphophallus Paeoniifolius on CNS depressant and antiepileptic activity is available in literature; therefore we undertake this project.
6.3 Objectives of the study:
1.To evaluate the CNS depressant activity of Amorphophallus paeoniifolius on experimental animals by using following models
  • Pentobarbitone induced sleeping time
  • Exploratory activity
2. To evaluate the Antiepileptic activity of Amorphophallus paeoniifolius on experimental animals by using following models
  • MES induced seizures
  • Picrotoxin induced seizures
MATERIALS AND METHODS
7.1 Source of data:
  • Laboratory experiments on animals.
  • Literature survey
  • Books and journals
  • J-Gate@Helinet(RGUHS)
7.2 Method of collection of data:
Materials :
  • Collection, identification and authentification of the plant by a Botanist.
  • Chemicals and Drugs: Ethanol 90%, Picrotoxin, Chlorpromazine, Pentobarbitone.
Extraction:
Shade dried powdered tuber of Amorphophallus paeoniifolius will be extracted with ethanol in a Soxhlet’s apparatus at 60-80o C. The extract will be concentrated to a smaller volume andevaporated to dryness in vacuum desiccators to yield ethanolic extract of Amorphophallus paeoniifolius.
Animals:
Albino Wistar rats of either sex weighing 150-200g and swiss albino mice of either sex weighing 20-30g will be used. The animals will be maintained under controlled conditions of temperature (23 ± 2C), and 12-h light-dark cycles. The animals will be randomized into experimental groups and housed in sanitized polypropylene cages containing sterile paddy husk as bedding. They will have free access to standard pellets as basal diet and water ad libitum.
Effect of ethanolic extract of Amorphophallus paeoniifolius on CNS depressant activity:
(i)Pentobarbitone induced sleeping time13:
Mice of either sex will be randomly allocated to the different control and test groups. They will be treated with ethanolic extract of Amorphophallus paeoniifolius and pentobarbitone sodium (40 mg/kg, i.p.) will be administered 30 min later. The control group receive 10 ml/kg normal saline, i.p. 15 min before pentobarbitone. For positive control group; pentobarbitone (40 mg/kg, i.p.) will be administered 15 min after chlorpromazine hydrochloride (1 mg/kg, i.m.). Onset of sleep will be taken as the time when mice accept the decubitodorsal position for three consecutive trials. Conversely, the duration will be considered complete when mice do not accept the decubitodorsal position for three consecutive trials.
(i)Exploratory activity13:
This study will be carried out by the whole-board method. Each mouse will be placed at one corner of the board and the animal moved about and dipped its head into the holes indicating exploratory behavior. The number of dips in 7.5 min will be recorded. The test will be carried out 30 min after oral treatment with ethanolic extract or chlorpromazine (1 mg/kg, i.m) to different groups of mice.
Effect of ethanolic extract of Amorphophallus paeoniifolius on Antiepileptic activity:
(i)MES induced seizures14:
Mice will be administered ethanolic extract of Amorphophallus paeoniifolius and 30 min later, Maximal electroshock seizures will beelicited by alternating current of 50 mA intensity delivered for 0.2 sec via corneal electrodes. Phenytoin will be used as standard drug (25 mg/kg, b.w) and 30 min later seizures were administered. Abolition of the hind limb tonic extension component of the seizure will be defined as protection.
(ii)Picrotoxin-induced seizures13:
In this test, picrotoxin (6 mg/kg, i.p) will be used to induce seizure. Adult wistar rats will be treated with ethanolic extract, 30 min before picrotoxin. Phenytoin (25mg/kg) will be used as standard drug. Seizure stage and seizure latency will be evaluated.
The animals will be grouped as follows for carrying out above activities.
Group / Treatment
Group 1 (normal) / Distilled water
Group 2 (positive control) / Inducing agents(MES, Pentobarbitone, Picrotoxin)
Group 3 (compound-I lower dose) / Test drug + inducing agent
Group 4 (compound-I higher dose) / Test drug + inducing agent
Group 5 (Standard) / Standard drug + inducing agent
Statistical analysis:
The data obtained will be expressed in MEAN ± SEM and analyzed by one way ANOVA followed by Tukey’s test.
7.3 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS /ANIMALS? IF SO PLEASE DESCRIBE BRIEFLY.
Yes, the above study requires investigation to be done on the rats and mice for the determination of CNS depression and antiepilepticactivities.
7.4 HAS ANIMAL ETHICAL COMMITTEE CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?
The copy of the ethical clearance of certificate obtained from Institutional Animal Ethical Committee (IAEC).
REFERENCES:
  1. Yu ZF, Kong LD, Chen Y. Antidepressant activity of aqueous extracts of Curcuma longa in mice: J Ethnopharmacol 2002;83:161.
  2. The World Health Report. Mental health: new understanding new hope. WHO, Geneva, 2001.
  3. Desari, Gandhi.Element of Pharmacology.14thed. Ahmedabad:Shah SB 2007.p. 281-9.
  4. Salil B, Parantasen, Arunaba R. Pharmacology.2nded. New Delhi:Elsevier 2005;247- 9.
  5. Sharma AK, Khosla R, Mehta VL, Kela AK. Anti epileptic agents newer generation.Ind J Pharmacol 1996;28:1-10.
  6. Nikalje APG, Ghodke M, Girbane A.GABA modulating agents: A brief review. Asian J Biol Sci 2011;4:201-20.
  7. Kivim M, Hamer M, Batty GD. Antidepressant medication use, weight gain, and risk of type 2 diabetes: a population-based study. Diabetes Care 2010;33(12):2611–6.
  8. Warrier PK and Ramankutty C. Indian Medicinal Plants. Delhi:Orient Longman; 1994.p. 132-7.
  9. Shankhajit DE, Dey YN, Ghosh AK. Anti-Inflammatory Activity of Methanolic Extract OfAmorphophallus Paeoniifolius and its Possible Mechanism. Int J Pharma Bio Sci 2010;1(3):23-35.
  10. Purwal L, Shrivastava V and JainUK. Studies On AntiDiarhhoeal Activity Of Leaves Of Amorphophallus Paeoniifolius In Experimental Animals. IntJ Pharm Sci Res 2011;2(2):468-71.
  11. Nataraj HN, Murthy RLN, Setty SR. In Vitro Evaluation OfGastro-Protective Of Suran-A Possible Explanation Through HPTLC Analysis. Int Res J Pharm 2011;2(9):103-06.
  12. Dey YN, GhoshAK. Evaluation of Anthelmintic Activity Of The Methanolic Extract Of Amorphophallus Paeoniifolius Tuber. Int JPhar Sci Res 2010;1(11):117-21.
  13. Dhanasekaran S, Palayan M. CNS depressant and antiepileptic activities of the methanol extract of the leaves of Ipomoea Aquatica Forsk. E-Journal Of chemistry 2010;7(4):1555-61.
  14. Aytemir MD, Calis U. Synthesis of Some New Hydroxypyranone Derivatives and Evaluation of Their Anticonvulsant Activities. FABAD J.Pharm Sci 2006;31:23-9.

9. / SIGNATURE OF THE STUDENT
10. / REMARK OF THE GUIDE
The above mentioned information and literature has been extensively investigated, verified and is found to be correct. The present study will be carried out under my supervision and guidance.
11. / 11.1 NAME AND DESIGNATION OF THE GUIDE
11.2 SIGNATURE / Dr. PREETI. KULKARNI M.Pharm.,Ph.D.
PROFESSOR,
DEPT. OF PHARMACOLOGY,
SET’s COLLEGE OF PHARMACY S.R.NAGAR, DHARWAD- 580002
11.3 CO-GUIDE
11.4 SIGNATURE / Mr. S M.BIRADAR. M.Pharm
LECTURER,
DEPT.OF PHARMACOLOGY,
S E T’s COLLEGE OF PHARMACY,
S. R. NAGAR,
DHARWAD-580002
11.3 HEAD OF THE DEPARTMENT
11.4 SIGNATURE / Dr. A R.Kulkarni M.Pharm.,Ph.D.
PROFESSOR
DEPT. OF PHARMACOLOGY,
Set’s College of Pharmacy
s.r.nagar, Dharwad-580002
12. / 12.1 REMARK OF THE PRINCIPAL / The above mentioned information is correct and I recommend the same for approval.
12.2 SIGNATURE / Dr. V H. Kulkarni M.Pharm.,Ph.D.
PROFESSOR AND PRINCIPAL,
Set’s College of Pharmacy
s.r.nagar, Dharwad-580002

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