Rajiv Gandhi University of Health Sciences Karnataka, Bengaluru

“EVALUATION OF ANTIHYPERTENSIVE, ANTIULCER AND ANTIDIARRHEAL ACTIVITY OF PLUMBAGO ZEYLANICA LIN”

A Protocol submitted to Rajiv Gandhi University of Health Sciences Karnataka, Bengaluru

In partial fulfillment of the requirement for the award of

MASTER OF PHARMACYIN

PHARMACOLOGY

Mr. SHAKEEL AHAMED KHAN

Department of Pharmacology,National College of Pharmacy,

Balraj-Urs Road,Shimoga-577 201Karnataka-INDIA

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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BENGALURU

Annexure – II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

01 Name and Address of the Candidate

Mr.Shakeel ahamed khanS/o Sardar khanNallur camp, Near Urdu SchoolChannagiri(T),Davanagere(D),Karnataka.

02 Name of the Institution

NATIONAL COLLEGE OF PHARMACY,BALRAJ-URS ROAD,SHIMOGA-577 201,KARNATAKA-INDIA.

03 Course of the StudyBranch M.PHARM. (PHARMACOLOGY)

04 Date of Admission to course 01-08-2013

05 Title of the Topic

"EVALUATION OF ANTIHYPERTENSIVE, ANTIULCER AND ANTIDIARRHEAL ACTIVITY OF PLUMBAGO ZEYANICA LINN''

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Brief resume of the intended work6.1. Need for the Study Enclosure – I

6.2. Review of the Literature Enclosure – II

6.3. Objective of the Study Enclosure – III

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Materials and Methods7.1. Source of data Enclosure – IV

7.2. Methods of collection of data Enclosure – V7.3. Does the study require any Investigations on animals? If yes give details

Enclosure – VI

7.4. Has ethical clearance been obtained from your institution in case of 7.3.

Enclosure – VI – A

08 List of References (About 4 – 6) Enclosure – VII

09 Signature of the Candidate

10 Remarks of the Guide The present research work is original and not published in any of the journals. This work can

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be carried out in our Department of Pharmacology, National College of Pharmacy, Shimoga.

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Name and Designation of (in Block Letters)

11.1. Guide

Mr. SHEKSHAVALI.T M. Pharma

LecturerNational College of Pharmacy,Balraj-Urs Road,Shimoga-577 201Karnataka-INDIA

11.2. Signature

11.4. Head of the Department Dr. I. J. KUPPAST M. Pharma, PhD, F.I.C.

Head of the DepartmentNational College of Pharmacy,Balraj-Urs Road,Shimoga-577 201Karnataka-INDIA

11.6. Signature

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Remarks of the Principal

12.1. Signature

The present research work is original and not published in any of the journals. This work can be carried out in our Department of Pharmacology, National College of Pharmacy, Shimoga.

Principal

Enclosure – I

Brief resume of intended work:3

6.1. Need for study:

Hypertension is one of the most common diseases affecting humans. It is

associated with morbidity and mortality worldwide. Hypertension is an

important public health challenge. Hypertension is a disease of complex

etiology, affecting 972 million people worldwide. It is estimated that the

worldwide prevalence of hypertension would increase from 26.4% in 2000

to 29.2% in 2025. Hypertension is an important risk factor for

cardiovascular and kidney diseases which has become a major global

burden on public health.[1]

Kidney diseases are increasingly prevalent public health concern. Renal

impairment is frequently associated with hypertension and there is

compelling evidence of the benefits of antihypertensive therapy for

reducing progression of kidney disease. The beneficial effects of

antihypertensive agents with their hemodynamic, antiproteinuric and

pleiotropic mechanisms provide a strong rationale for considering the use

of these agents in the treatment of high-risk patients.[2]

Diarrhoea is characterized by increased frequency of bowel movement,

wet stool and abdominal pain. Today it is a leading cause of malnutrition

and death among children in the developing countries of the world. Many

governments and international organizations are trying to control this

disease but the rate of incidence is still high, about 7.1 million per year.

Many synthetic chemicals like diphenoxylate, loperamide and antibiotics

are available for the treatment of diarrhoea but they have some side

effects. The natural drugs are used as antidiarrhoeal drugs, which are not

always free from adverse effects. Therefore, the search for safe and more

effective agents has continued to be an important area of active research.

Since ancient times, diarrhoea has been treated orally with several

medicinal plants or their extracts based on folklore medicine3.

Like diarrhoea, Peptic ulcer is the most frequent disorders of the

alimentary tract and in various countries its prevalence is estimated as 5-

10% of the adult population. This disorder remains one of the most

important problems, both in the practice of primary health care physicians

and gastroenterologist4. Ulcers are caused by an imbalance between

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aggressive factors (gastric acid, pepsin, Helicobacter pylori) and the

defensive factors (mucus, HCO3- and mucosal blood flow)5. Factors which

are responsible for pathogenesis of ulcers includes increased basal

secretory drive, increased parietal cell mass, increased post prandial acid

secretion, increased post prandial acid release, increased secratagogues,

decrease inhibition of acid, and gastrin secretion, rapid delayed gastric

emptying and decreased mucosal resistance. Genetic and environmental

factors are also thought to play a role6,7 .

Scientific Classification:

Family: Plumbaginaceae

Synonym: plumbago viscora, plumbago scandens

Class: Magnoleopsida

Subclass: Caryophyllidae

Species: Zeylanica

Vernacular Name: White Leadwort, while other vernacular names are as

follows: Chitraka (Hindi- Uttar Pradesh), Telhidak angouba (Manipur),

Chittiramoolam karimai (Tamilnadu), Vellakoduveli (Malayalam),

Chitramulika (Karnataka), Safaid-sitarak (West Bengal), Ogni (Orissa),

Chitrmulam (Andhra Pradesh), etc.

The plant Plumbago zeylanica linn is an evergreen small perennial

shrub which grows to a height of about 3–4 feet. The leaves are simple,

alternate, oblong, spirally arranged, hairy margin, thick, and flashy, 4–10

cm long, pointed at the tip. The flowering occurs from September to

November. The flowers are white in color, 10–25 cm long.8

Plumbago zeylanica linn are the roots which have been traditionally

reported to be used for a variety of ailments such as dyspepsia, piles, 5

diarrhoea, skin diseases and rheumatism9. The leaves are caustic, vesicant,

aphrodisiac, and good for scabies (yunani) 10

Chemical constituents

The naphthoquinones, plumbagin, composed naphthoquinones like

plumbagin, 3-biplumbagin, chloroplumbagin, chitranone, elliptone. The

coumarins seselin, 5-methoxyseselin, suberosin, xanthyletin and Other

compounds were 2,2-dimethyl-5-hydroxy-6-acetylchromene, plumbagin

acid, ß-sitosterol, ß-sitosteryl-glucoside, bakuchiol, 12-

hydroxyisobakuchiol, saponaretin, isoorientin,isoaffinetin, psorealen.Roots

of P. zeylanica: Two plumbagic acid glucosides, 3′-O-β-glucopyranosyl

plumbagic acid11.

Enclosure – II

6.2. Review of literature:

1. Rajesh Kumar, Sushil Kumar, Arjun patra, S.Jayalaxmi carried out the

studies on evaluation of hepatoprotective activity of aerial parts of

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Plumbago zeylanica linn and the result concluded that the methanol

extract of the aerial parts of Plumbago zeylanica possess significant

hepatoprotective activity.12

2. Edwin S, Joshi SB, Jain DC, carried out studies on Antifertility activity of

of leaves of Plumbago zeylanica linn in female albino rats and the result

have an Antifertility activity.13

3. Kakjing Radul Falang, Mary ogonnaya uguru, noel nenman wannang,

Iliya hosea azi, Nwoye chamaka carried out studies on antiulcer activity of

the aqueous extract of the roots of Plumbago zeylanica linn against

Indomethacin and asprin induced ulcer in Albino rats and provides a

bassis for its ethono medicinal uses in treatment of dyspepsia.14

4. Agbaje E.O. and Adeniram J.O. carried out the studies on some

gastrointestinal effect of the aqueous root extract of pulmbago zeylanica

(lead Wort). The present study has substantial folkloric use of PLZ in both

diarrhea and gastric ulcerations.15

5. Vishnukanta, A.C. Rana carried out the studies on evaluation of

anticonvulsant activity of Plumbago Zeylanica linn. Finally its concluded

that this hydro alcoholic extract of leaves of plumbago zeylanica linn has

no anticonvulsant activity.16

6. N. Kanchana and A.Mohamed Sadiq studied the hepatoprotective effect of

plumbago zeylanica on paracetamol, induced liver toxicity in rats. The

present study reveals that the petroleum ether root extract of plumbago

zeylanica could afford and significant protection against paracetamol

induced hepatocellular injury.17

7. Desai.H.P., Kapadia.M.D., Kharat.A.R. studied that evaluation of

anthelmintic activity of plumbago zeylanica linn was observed by

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gradually increasing the dose of extract. Methanolic extract of plumbago

zeylanica linn showed higher activity as compared to water extract.18

8. Sunil C., Duraipandiyan V., Agastian P., Iqnacimuthu S. Studied the

antidiabetic effect of plumbagin iolated from plumbago zeylanica linn and

its effect on GLU T4 translocation in steptozotocin-induced diabetic rats.

The results indicated that plumbagin enhenced GLU T4 translocation and

contributed to glucose homeostasis. It could be further probed for use as a

drug to treat diabetes.19

9. Vineet mittal, S K Sharma,. Deepak koushik,. Meenu khatri,.and Kusum

tomar carried out the the comparative study of analgesic activity of

plumbago zeylanica linn. Callus and root extract in experimental mice and

the result has shown the significant peripheral analgesic activity.20

10. Tilak JC,. Adhikari S,. Devasagayam,. Carried out the studies on

antioxidant properties of plumbago zeylanica linn. An Indian medicinal

plant and its active ingredients, plumbagin and result concluded that

extract of plumbago zeylanica linn and its ingredient plumbagin have

significant antioxidant activity.21

Enclosure – III

6.3. Objectives of study:

1. Collection and authentication of Plumbago zeylanica linn.

2. Drying of whole of Plumbago zeylanica linn.

3. Extraction of the plant using different solvents.8

4. Phytochemical investigations of extracts.

5. Screening of antihypertensive, antiulcer and antidiarrhoeal activity.

6. Evaluation of Toxicological studies.

Enclosure – IV

MATERIALS AND METHODS:

7.1. Source of data:

The required data will be obtained from:

1. Electronic data [internet].

2. Published Research Papers.

3. Review and Research Articles from Journal.

4. Library, National College of Pharmacy, Shimoga, Karnataka, India.

Enclosure –V

7.2. Method of collection of data:

1. The plant of Plumbago zeylanica linn will be collected from local areas of

Shimoga district, Karnataka.

2. Screening of Pharmacological activities – Antihypertensive, Antiulcer and

antidiarrhoeal activity.

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3. LD50 studies.

Antihypertensive activity will be carried out with male Wister rats,

mean arterial BP was measured at different time intervals. Activity will be

concluded by comparing the results of control, standard and test samples.

Antiulcer activity: Evaluation of the extracts for anti-ulcer activity

studies. Screening methods are adopted from standard books and from

national and international scientific journals. Method of collection of data

(including sampling procedure) Data will be collected from experimental

studies and subjected to statistical analysis for antiulcer activities.

1. Aspirin induced ulcer.

2. Alcohol induced ulcer.

3. Pylorus ligation induced ulcer

Antidiarrhoeal: 1.Caster oil induced diarrhea, 2. Charcoal meal test

Toxicological studies will be carried out as per the standard procedure

(as per OECD guidelines) and the results are compared with treated and

control group. The results will be analyzed by ANOVA test.

Enclosure – VI

7.3. Does the study require any investigation or intervention to be

conducted on patients or other humans or animals?

As per the standard procedure, Antihypertensive, antiulcer and

antidiarrheal activities of areal parts of Pumbago zeylanica Linn will be

carried out on the Wistar albino mice and rats.

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Enclosure-VI A

7.4. Has ethical clearance been obtained from your institution?

Ethical clearance is provided by the Institution.

Clearance number: NCP/IAEC/CL/237/2013-14.

Enclosure –VII

List of references:

1. Kearney PM, Whelton M, Reynolds K, Muntner P, Whelton PK, He J.

Global burden of hypertension: analysis of worldwide data. Lancet 2005;

365(9455):217-23.

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2. Ruilope LM. Angiotensin receptor blockers: RAAS blockade and

renoprotection. Curr Med Res Opin. 2008; 24(5):1285-93.

3. Ezekwesili CN, Obiora KA, Ugwu OP. Evaluation of antidiarrhoeal

property of crude Aqueous Extract of Ocimum gratissimum L. (Labiatae)

In Rats. Biokemistr, 2004; 16(2): 122-131.

4. Schabowski J., Pitera J. “Peptic ulcer among rural population in a selected

region of south-eastern Poland”, Ann Agric Environ Med 2004; 11: 323-

327.

5. Takeuchi K, Akira K, et al. Effects of pantoprazole, a novel H+/K+-

ATPase inhibitor,on duodenal ulcerogenic and healing responses in rats: A

comparative study with omeprazole and lansoprazole. J Gastroent Hepat

1999; 14: 251-257.

6. Feldman M.,Scharschmidt B.F.,Sleiscnger M.H., Gastrointestinal and

Liver disease pathophysiology diagnosis /management.6th edition volume

I, McGraw-Hill, Inc, Philadelphia London Toranto Montreal Sydney

Tokyo; 251-327.

7. Harsh mohan., Text book of Pathology, 4th edition, New Delhi; Jaypee

Brothers, 2000: 511-569.

8. Geltz NR, Russell SD. Two-Dimensional Electrophoretic Studies of the

Proteins and Polypeptides in Mature Pollen Grains and the Male Germ

Unit of Plumbago zeylanica. Plant Physiol 1988; 88:764-9.

9. Onfade DD, jewell AP, okeshina AB, Yong k, ojele M, Nwanze JC,.

International journal of biochemistry of bioinformatics 2012;1(5):124-

130.

10. K R kirtikar and B D basu, Indian medicinal plants, 1991 vol 2, 2nd

edition. Dehra Dun. Shiva offset press; p 1466-1467.

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11. Lie-Chwen Lin, Ling-Ling Yang, Cheng-Jen Chou,Cytotoxic

naphthoquinones and plumbagic acid glucosides from Plumbago

zeylanica linn, Phytochemistry, 2003;62(4): P 619-622.

12. Rajesh kumar, Sushil Kumar, Arjun Patra, S. JayaLaxmi, Studies on

hepatoprotective activity of areal parts of plumbago zeylanica linn.

International journal of pharmacy and pharmaceutical sciences 2009;1(1)

13. Edwin.S., Joshi.S.B., Jain.D.C. The leaves of plumbago zeylanica linn,

acetone and ethanolic extracts of leaves of plumbago zeylanica have an

anti fertility activity.2009;14(3):233-239.

14. Kakjing Radul Falang, Mary ogonnaya uguru, noel nenman wannang,

Iliya hosea azi, Nwoye chamaka carried out studies on antiulcer activity of

the aqueous extract of the roots of Plumbago zeylanica linn. Scholars

research library, 2012;2(5):563-567.

15. Agbaje.E.D. and Adeniran.J.O carried out the studies on some

gastrointestinal effect of the aqueous Root extract of plumbago

zeylanica(lead Wort). African journal of Bio medical research, 2009;

12(1):63-68.

16. Vishnukanta, A.C. Rana evaluation of anti convulsant of plumbago

zeylanica linn 2010; 3(1):76-78.

17. N. Kanchana and A.Mohamed Sadiq. Hepatoprotective effect of

plumbago zeylanica linn on paracetamol induced liver toxicity in rats.

International journal of pharmacy and pharmaceutical sciences 2009;

3(1):151 to 154.

18. Desai.H.P., Kapadia.M.D. Kharat.A.R. evaluation of anthelmintic activity

of plumbago zeylanica linn. 2012;3(11):4281.

19. Sunil C., Duraipandiyan V., Agastian P., Iqnacimuthu S. Studied the

antidiabetic effect of plumbagin iolated from plumbago zeylanica linn. U 13

S nationl library of medicine national institute of health 2012;

50(12):4356-4363.

20. Veneet Mittal, SK sharma, deepak kaushik, meenu katri and kousum

tomar a comparative study of analgesic activity of plumbago zeylanica

linn. callus and root extract in experimental mice . Research journal of

pharmaceutical biological and chemical sciences. 2010; 1(4):830-836.

21. Tilak JC, Adhikari S,. Devasagayam TP. Studies on antioxidant properties

of plumbago zeylanica linn an indian medicinal plant and its active

ingredient. US national library of medicine national institute of health

2004; 9(4) p: 219-227.

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