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Study on phytochemical and neuropharmacological effects of cassia tora seeds. M. Pharm dissertation protocol submitted to the


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STUDY ON PHYTOCHEMICAL AND NEUROPHARMACOLOGICAL EFFECTS OF CASSIA TORA SEEDS.
M.PHARM DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,

BANGALORE.


By

MOHD. INTEKHAB ALAM

B.Pharm.
Under the guidance of
Dr. D.K.SURESH

M.pharm., Ph.D.



Professor and Head

Dept. of Pharmacology
http://luqmanpharmacyglb.org/images/image002.jpg
DEPARTMENT OF PHARMACOLOGY

LUQMAN COLLEGE OF PHARMACY, GULBARGA

2013-2014

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION


1.

Name of the Candidate and Address (In block letters)

MOHD. INTEKHAB ALAM

Room No-38, Boys Hostel LCP

2.

Name of the Institution

Luqman College of Pharmacy, gULBARGA – 585 102

3.

Course of Study and Subject

M.Pharmacy (PharmacOLOGY)

4.

Date of Admission to Course

14/06/2013

5.

Title of the Topic

STUDY ON PHYTOCHEMICAL AND NEUROPHARMACOLOGICAL EFFECTS OF CASSIA TORA SEEDS.

6

BRIEF RESUME OF THE INTENDED WORK




6.1 Need for the Study

Neuropharmacology is a branch of neuroscience involving the study of drugs that alter the nervous system and its functioning, specifically within the brain. The goal of neuropharmacology in general is to understand the basic functioning of impulses and signals within the brain in order to discover ways in which drugs can be used to treat neurological disorders and drug dependence 1. 21st Centaury is called the centaury of neurological disorders. Neurological related problems are so common today, that approximately 18% population suffer from disorders each year 2

Neurological disorders are disorders that can affect the central nervous system, peripheral system and autonomic nervous system. These disorders produces serious health problems like behavioral / cognitive syndrome, sleep disorders, peripheral disorders, epilepsy, neurodegenerative disorders, parkinsonism, neoplasm and many others3

In spite of the recent advances in understanding neurological disorders, there are still no effective therapies. Because in modern medicine treatment is empirical symptom oriented and not disease specific. And also the currently available drugs which are in clinical use have characteristic profile and side effects4

Hence management of neurological disorders with agents devoid of any side effect is still a challenge to the medical system. This has led to an increase in the demand of natural products with neurological activity having lesser side effects. Many natural products having neurological effect have been discovered and used by man since prehistoric times4 for instance,

The methanolic leaf extract of Cissus cornifolia has got remedy against mental derangement 5 .

The aqueous extract of Ficus sycomorus stembark possesses a sedative and anticonvulsive properties6.

Myristica fragrans seeds possesses anxiogenic, sedative and analgesic activity7

Like above one of the most commonly available plant Cassia tora Linn. Belongs to family leguminacea is a shrub, extensively used in traditional medicine in tropical and warm subtropical countries. Cassia tora commonly found in waste grounds and secondary forest. Parts of Cassia tora are used as an antifungal, antihelmintic, diuretic, expectorant, laxative, treatment of glaucoma, hypertension, treatment of skin diseases, ringworm and itch8. Also the seeds of Cassia tora are used to improve visual activity in Chinese medicine and it is also reputed for its medicinal values as antiasthemic and diuretic agents9. In India the plant is used in the treatment of snakebites and scorpion stings10

Through literature survey reveals the presence of saponins and flavonoids in Cassia tora11 The saponins and flavonoids have been reported to be responsible for neuropharmacological effects. These results are supported by several researchers12

To the best of our knowledge the neuropharmacological effects of Cassia tora were not been explored yet. In the light of literature survey which explored the presence of saponins and flavonoids in cassia tora and also based on the knowledge of traditional use of this plant as an analgesic and anticonvulsant in folk medicine practice, an attempt could be made to evaluate its neuropharmacological effects in experimental animals using various scientific animal models.

6.2 Review of the literature

Literature survey reveals that, the human nervous system is an extremely complex structure, having more than 12 billion nerve cells. Together with the endocrine system







6.2 Review of the literature

Literature survey reveals that, the human nervous system is an extremely complex structure, having more than 12 billion nerve cells. Together with the endocrine system it coordinates and regulates the functioning of all body organs13 .

Central nervous system or psychiatric disorders are the serious health problems. Despite introduction of neurological agents from natural and synthetic sources neurological disorders and its complication continued to be a major problem in the world population. How ever synthetic medicines are available in the treatment of neurological disorders but having large number of side effects4 . Indian traditional medicine is one of the richest medicinal systems, among those available around the world.

Literature survey indicates that the petroleum and chloroform extracts of Laportea crenulata Roots has got potent CNS Stimulant activity 14 Similarly petroleum ether extract of Peperomia pellucida seeds in the treatment of mental disorder15

Literature review also shows that good piece of work has been done on various pharmacological activities of Cassia tora. But there are no reports on its neuropharmacological effects. Some of the published reports on pharmacological and phytochemical aspects of Cassia tora are listed as below:


  1. 1. Hepatoprotective effects of Cassia tora seeds16.

  2. 2. Antinociceptive and smooth muscle contracting activities of the methanolic extract

  3. of Cassia tora seeds.17

  4. 3. Leaves of Cassia tora as a novel cancer therapeutic –An in vitro study.18

  5. 4. Immunostimulantory activities of Cassia tora.19


6.3 Objectives of the Study

The literature review reveals that Neuropharmocological activity of Cassia tora seeds has not been reported. In view of this, the present study is aimed to investigate the phytochemical and Neuropharmocological activity of various extracts of Cassia tora seeds in different experimental animal models.





MATERIALS AND METHODS

7.1 Source of Data

The whole work is aimed to generate data from the laboratory that is experiments on animals. Albino rats and mice will be used for this purpose.

The scheme of proposed work is as follows.


  1. Collection of plant material.

  2. Identification and authentication of the plant material

  3. Shade drying and powdering of the plant material.

  4. Soxhlet extraction of powdered leaves from suitable solvents systems.

  5. Preliminary phytochemical investigation of various extracts.

  6. Study of the acute toxicity for determination of LD50 of the extract in mice.

  7. To evaluate the Neuropharmocological effect of Cassia tora leaf extracts in rodents.


7.2 Methods of Collection of Data
1. Phytochemical studies:

The seeds of Cassia tora were dried in shade and processed to a coarse powder. The coarse powder is percolated with successive extraction in increasing polarity as explained by Kokate20. The solvents used are petroleum ether, chloroform, alcohol and distilled water. The crude extracts obtained by removing the solvent will be subjected to preliminary phytochemical screening, some extracts are used for the pharmacological investigations. The detailed methodology is as under.



Acute Toxicity test:

It is planned to study the acute toxicity of solvent extracts of Cassia tora in albino mice of either sex (25-30 gms). Fixed dose methods OECD guidelines 420 is adopted for toxicity study.



Exploratory behavior:

Albino mice were devided into six groups containing ten animals each. The test was carried out 30 minutes after pretreatment of the various extracts of cassia tora seeds and vehicle. Chlorpromazine hydrochloride (1mg/kg i.p.) was used as standard control drug. In this method a white printed wooden board (40cm×40cm) with four equidistant holes (1cm diameter×2cm depth) is used. The mice were placed at the center of the board and moved freely in the box. A head dip into holes was used to indicate exploratory behavior. The number of dip was observed for ten minutes21.


Analgesic activity:

Hot plate method:

The albino mice (n=5) devided into 6 groups were placed individually on a hot plate maintained at 55º±1º C, after 0, 30, 60, 90, 120 and 150 mins of administration of vehicle, various extracts of Cassia tora seeds and pentazocine (10mg/kg). and the time required to lick the paws was measured for each mouse. The percentage of analgesia was calculated.



Acetic acid induced writhing:

Albino mice devided into groups of six (n=10) received various extracts of Cassia tora seeds, Ibuprofen(40mg/kg orally) and vehicle 30mins before intraperitonial injection 0.1ml of 0.6% solution of acetic acid. The number of abdominal constrictions (writhing) was measured for 20mins and percentage analgesia was calculated7.



Hexobarbitone induced sleeping time:

The sedative- hypnotic effect of the extracts was assessed using hexobarbitone (85mg/kg i.p.) induced sleeping time in mice. Albino mice were devided into five groups with 10 animals per groups and later treated with different extracts of Cassia tora seeds and the control , 30 mins later animals were given hexobarbitone. Sleeping time was measured as a time interval between loss and regain of righting reflex 21.


Maximal electroshock seizure (MES) test:

A total 36 mice were divided into 6 groups and alternating current of 50Hz and 150mA was delivered to experimental animals through bicorneal electrodes for 0.2 seconds. A drop of 0.9% saline solution was poured into both eyes prior to placing the electrodes. The different extracts of Cassia tora seeds as well as Phenobarbital were injected intraperitonially 45 mins before induction of MES and control group will receive only vehicle. Duration of tonic convulsion and the percentage of seizure protection and mortality were recorded22.







INCLUSION CRITERIA

Normal and healthy animals weighing between 150-180 gms for rats and 25-30gms for mice will be included in the study.



EXCLUSION CRITERIA

The albino rats and mice which do not fall the above mentioned weights are excluded from study.


STATISTICAL ANALYSIS

All values will be expressed as mean±SEM from any group. Results will be subjected to statistical analysis using one- way ANOVA and allowed by post hoc test (Dunnet ‘T’ test). p<0.05 will be considered as statistical significant.




7.3 Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please describe briefly.

Yes, the above study requires investigation on animals like albino rats and mice.





7.4 Has ethical clearance been obtained from your Institution in case of 7.3?

Yes, the study is cleared from institutional animal ethics committee (IAEC copy enclosed).







8.


LIST OF REFERENCES




  1. Everitt B. J and Robbins T. W. Neural systems of reinforcement for drug addiction from actions to habits to compulsion. Nature Neuroscience 2005; 8(11):1481-1489.

  2. www.phobias.about.com/od/prevalence/a/phobprevus

  3. http://en.wikipedia . org/wiki/The__Neurologist

  4. Tripathi KD. Essential medical pharmacology. 2nd Edition. New Delhi. Jaypee brothers publication 1988; 388-389p.

  5. Musa A. M, Yaro A. H, Usman H, Magaji M.G and Habu M. Phytochemical and some Neuropharmacological studies on the Methanolic Leaf Extracts of Cissus cornifolia in Mice. International Journal of Pharmacology 2008;4(2): 145-148p.

  6. Umar Kyari Sandabe, Patrik Azubuike Oneyili and Gregory Anene Chibuzo. Sedative and anticonvulsant effects of aqueous extract of Ficus sycomorus L. steambark in rats. Veterinarski archive 2003; 73(2):103-110

  7. Ganeshchandra Sonavane, Vikram Sarveiya, Veena Kasture, Sanjay B. Kasture. Behavioral actions of Myristica fragrans seeds. Indian Journal of Pharmacology 2001; 33: 417-424.

  8. Ahmad Z, Somchit M.N, Mutalib AB and Norli S. In vitro antifungal properties of Cassia tora extracts. Proceeding of the Regional Symposium on Environment and Natural Resources 2002;1: 472-476.

  9. Jae Sue Choi, Hee Jung Lee and Sam Sik Kang. Alaternin, Cassiaside and Rubrofusarin gentiobioside, Radical Scavenging Principles from the Seeds of Cassia tora on 1,1-Diphenyl-2-picrylhydrazyl (DPPH) Radical. Arch. Pharm. Res 1994;17(6):462-466.

  10. Ali Mohmoud El-halwany, Mi Hwa Chung, Norio Nakamura, Chao-Mei MA, Tsutomu Nishihara, and Masao Hattori. Estrogenic and Anti-estrogenic Activities of Cassia tora Phenolic Constituents. Chem. Pharm. Bull 2007; 55(10):1476-1482.

  11. Roopashreen TS, Raman Dang, Shobha Rani RH, Narendra C. Antibacterial activity of antipsoriatic herbs: Cassia tora, Momordica charantia and Calendula officinalis. International Journal of Applied Research in Natural Products 2008; 1(3):20-28.

  12. Viswanatha Swamy A.H.M, Thoppeswamy A.H.M, Manula D.V. and Mahendra kumar C.B. Some Neuropharmacological studies on the Methanolic Leaf Extracts of Cissus Quadranfularis in mice. African Journal of Biomedical Research 2006;9:69-75.

  13. Barar FSK. Essentials of Pharmacotherapeutics, 3rd edition. S.Chand publishers. New Delhi. 1999.62p.

  14. Alam Khan, Ashik Mosaddik, Mukhlesur Rahman, Motiur Rahman and Ekramul Haque S. S. Jahan, M. S. Islam, Sohel Hassan. Neuropharmacological effects of Laportea crenulata Roots in mice. Journal of Applied Sciences Research 2007;3(7):601-606.

  15. Khan A, Rahman M. Islam M.S. Neuropharmacological effects of Peperomia pellucida leaves in Mice. DARU 2008;16(1):35-40.

  16. Vetrivel Rajan A, Shanmugavalli N, Greety Sunita C. and Umashankar V. Hepatoprotective effects of Cassia tora on CCl4 induced liver damage in albino rats. Ind. J. of Sci. and Tech 2009;2(3):41-44.

  17. Chidume F.C, Kwanashie H.O, Adekeye J.O, Gamaniel K.S. Antinociceptive and smooth muscle contracting activities of the methanolic extract of Cassia tora leaves. Journal of Ethanopharmacology 2002;81:205-209.

  18. Rejiya C.S, Cibin T.R, Annie Abraham. Leaves of Cassia tora as a novel cancer therapeutic –An in vitro study. Toxicology in vitro 2009;23:1034-1038.

  19. Jaw-Ming Cherng, Wen Chiang, Ji-Hung Wang. Anthraquinines of edible wild vegetable and immunostimulantory activities of Cassia tora. Food Chemistry 2008;107:1576-1580.

  20. Kokate CK. Practical Pharmacognosy. Pune. Nirali Prakashan 2006. 107-111p.

  21. Wakeel OK, PI.Aziba, Ashorobi R.B, Umuloro S, Dderibigbe A.O and Awe E.O. Neuropharmacological activities of Ficus platyphylla stem bark in mice. African Journal of Biomedical Research 2004;7: 75-78.

  22. Hossein Hosseizadeh and Vahid Khosravan. Anticonvulsant effects of Aqueous and Ethamolic extracts of Crocus sativus L. stigmas in mice. Arch Irn Med 2002;5(1):44-47.







9.

Signature of Candidate


MOHD. INTEKHAB ALAM

10.

Remarks of the guide

The work is highly justifiable and is feasible to work in the institution.



11.

Name & Designation of (in block Letters )

M.PHARMACY STUDENT




11.1 Guide

Dr. D K Suresh

M Pharm., Ph.D.

PROFESSOR & HEAD

DEPT. OF PHARMACOLOGY,

LUQMAN COLLEGE OF PHARMACY,

GULBARGA.






11.2 Signature







11.3 Co-guide


---------




11.4 Signature




12.

12.1 Remarks of the

Chairman & Principal


We will provide all the necessary facilities required for the proposed research work.



So, recommended for registration




12.2 Signature




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