Rajiv Gandhi University of Health Sciences Karnataka, Bengaluru

“STUDIES ON THE PHARMACODYNAMIC AND PHARMACOKINETIC INTERACTIONS BETWEEN BIOFLAVONOIDS AND CALCIUM CHANNEL

BLOCKERS”

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

O.PRIYANKA

Department of Pharmacology,National College of Pharmacy,

Balraj-Urs Road,Shimoga-577 201

Karnataka-INDIA

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BENGALURU

Annexure – II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

01 Name and Address of the Candidate

O.PRIYANKA,D/O O.APPALARAJU,DOOR NO. 30/5/35,KRISHNA GARDENS,DABA GARDENS, VISHAKAPATNAM, A.P-530020

02 Name of the Institution

NATIONAL COLLEGE OF PHARMACY, BALRAJ-URS ROAD,SHIMOGA-577201,KARNATAKA, INDIA

03 Course of the StudyBranch M. Pharm. (Pharmacology)

04 Date of Admission to course 15-11-2010

05 Title of the Topic

STUDIES ON THE PHARMACODYNAMIC AND PHARMACOKINETIC INTERACTIONS BETWEEN BIOFLAVONOIDS AND CALCIUM CHANNEL BLOCKERS

06

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

07

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 be carried out in our Pharmacology Department laboratory.

11

Name and Designation of (in Block Letters)

11.1. Guide

11.2.Signature

11.3.Co-Guide (if any)

11.4.Signature

11.5. Head of the Department

11.6. Signature

Dr. I. J. KUPPAST M.PHARM, Ph.D, FIC.

PROFESSORDEPARTMENT OF PHARMACOLOGY,NATIONAL COLLEGE OF PHARMACY,SHIMOGA-577201KARNATAKA, INDIA

N.A.

N.A.

Dr. K. L. MANKANI M.Sc.,Ph.D

HEAD OF THE DEPARTMENT,DEPARTMENT OF PHARMACOLOGY,NATIONAL COLLEGE OF PHARMACY,SHIMOGA-577201KARNATAKA, INDIA

12

Remarks of the Principal

12.1. Signature

The present study is permitted to perform in the Pharmacology Department laboratory of our Institution and the study protocol has been approved by IAEC.

Principal

Enclosure – I

Brief resume of intended work:

Need for study:

Herbs have been used for medicinal purposes since the beginning of recorded

time. Although most people in the United States believe that herbs are harmless plants,

about one third of our drugs (including digitalis, morphine, atropine, and several

chemotherapeutic agents) were developed from plants1.

Most of the plants contain bioflavonoids as major active constituents.

Flavonoids or bioflavonoids (from the Latin word flavus meaning yellow), also

collectively known as Vitamin P and citrin3 are a class of plant secondary metabolites or

yellow pigments having a structure similar to that of flavones4.

Flavonoids commonly interact with most of the drugs and they claim to have

effect on CVS and blood vessels. Therefore, when bioflavonoids are taken concurrently

with antihypertensive drugs, interactions are possible1.

When herbal therapies and drugs are used together, they can interact in our

body, causing changes in the way the herbs and/or drugs function. Such changes are

called drug-herb interactions. They can be beneficial or harmful, depending on type of

interactions2.

Flavonoids have been used as effective constituents of several pharmaceuticals

used for treatment of capillary fragility and phlebosclerosis. The activities of certain

flavonoids inhibit capillary permeability5-8. Flavanoid namely Rutin inhibits platelet

aggregation, as well as decreasing capillary permeability, making the blood thinner and

improving circulation9. Cordio dichotoma Forst.f. Fruits belonging to the family

Boraginaceae is medium sized tree with a short, usually crooked trunk of 3-4ft. in

grith10, containing bioflavonoids have been selected for its interaction with anti-

hypertensives like calcium channel blockers in the study.

Hypertension (HTN) is elevated blood pressure. Conventional anti-hypertensive

drugs are usually associated with many side effects. About 75 to 80% of the world

population use herbal medicines, mainly in developing countries, for primary health

care because of their better acceptability with human body and lesser side effects11.

In the present study bioflavanoids needs to be coupled with selective

antihypertensive drugs like calcium channel blockers and more scientific research

needs to be done to verify the effectiveness, and elucidate the safety profile of such

herbal remedies for their antihypertensive potential.

Calcium channel blockers are the first line drugs in the treatment of

hypertension. Hence it is claimed to study the calcium channel blockers in the present

study.

Calcium is necessary for excitation-contraction coupling in both skeletal and

smooth muscles. Skeletal muscles have contractile activity whereas cardiac and

vascular smooth muscles contraction depends on extra cellular calcium concentration.

Mechanism of action of calcium channel blockers:

Calcium channel blockers mainly act by inhibiting the entry of calcium from L

channel into myocardial and vascular smooth muscles thus decreasing the availability

of intracellular calcium.

Pharmacodynamics actions of calcium channel blockers on peripheral blood

vessels: Calcium channel blocker relaxes the vascular smooth muscles in systemic as

well as pulmonary arterial circulation. They decrease vascular resistance and B.P in

both pulmonary and systemic circulation12-13.

Number of studies has been reported on calcium channel blockers and

bioflavonoids. However the interactions between bioflavonoids and calcium channel

blockers have not been documented in scientific literature. Hence the present study is

aimed at investigation of pharmacodynamic and pharmacokinetic interactions between

bioflavonoids and anti-hypertensive drugs like calcium channel blocker.

Enclosure – II6.2 Review of Literature:

1) The Bioflavonoid Galangin Blocks Aryl Hydrocarbon Receptor Activation and

Polycyclic Aromatic Hydrocarbon14.

2) Mechanisms for the Inhibition of DNA Methyltransferases by Tea Catechins

and Bioflavonoids15.

3) Bioflavonoids- Their pharmacokinetic interaction with cytochrome P450

isozymes and P-glycoprotein16.

4) Interaction of calcium channel blockers (CCB’s) with Noradrenaline and

Terbutalin on isolated right ventricle from normal and diseased rats17.

5) Bioflavonoids Classification, Pharmacological, Biochemical Effects and

Therapeutic Potential18.

6) Cardiac effects of calcium channel blockers in human volunteers19.

Enclosure – III

6.3 Objectives of present study:

1) The present study is planned to investigate the Pharmacodynamic and

Pharmacokinetic Interactions between Bioflavonoids and Calcium Channel

Blockers in rats.

2) In view of this some of the flavonoids have been isolated from selected plant

fruits.

3) The isolated flavonoids from the plant subjected to phytochemical analysis,

TLC, Column chromatography and elucidation of structures by spectral

analysis.

4) The selective calcium channel blockers will be obtained from the market.

5) The identified flavonoids and selected calcium channel blockers are subjected

for pharmacokinetic and pharmcodynamic study.

6) This study shows drug- herb interaction.

Enclosure – IV

Materials and methods:

7.1 Source of data:The required data will be obtained from:

1. Electronic data (Internet).

i. Online Journals

ii. Wikipedia.in

iii. Google

2. Published research papers.

3. Review and research articles from Journals.

4. Library, National College of Pharmacy, Shimoga.

Enclosure – V

7.2 Methods of collection of data:

1) In the present study flavonoids will be isolated from Cordia dichotoma Forst.f

fruit and used for the study.

2) The plant will be identified and authenticated with the help of botanist.

3) Plants are collected and subjected for extraction and fractionation.

4) Fractionated compound will be subjected to phytochemical analysis, TLC,

Column chromatography and elucidation of structures by spectral analysis.

5) Procurement of calcium channel blockers like Verapamil, Nifedipine and

Amlodipine from local market.

6) The identified flavonoids and selected calcium channel blockers are subjected for

pharmacokinetic and pharmcodynamic study.

7) Results are analysed by ANOVA test.

Enclosure – VI

7.3 Does the study require any investigation or intervention to be conducted on patients

or other humans/animals?

As per the standard procedure, to study pharmacodynamic and pharmacokinetic

interactions between bioflavonoids and calcium channel blockers will be carried out on the

wistar albino rats.

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/24/05/2011-12.

Enclosure – VII

List of References:

1) Eisenberg D M, Davis R B, Ettner S L, et al. Trends in alternative medicine use in

the United States: results of a follow-up national survey. JAMA, 1998. 280:

p.1569-1575.

2) Eisenberg D M and Kaptchuk T J. The Risk-benefit profile of commonly used

herbal therapies: ginko, st.John’s Wort, Ginseng, Eichinacea, Saw Palmetto and

Kava. Annals of internal medicine, 2002. 136: p.42-53.

3) Vitamin P, dictionary results, http://dictionary.reference.com/borwse/vitamin+P

4) Flavonoids (isofloavnoids and neoflavonoids),

http://www.iupac.org/goldbook/FO2424.pdf, IUPAC Compendium of chemical

terminology.

5) Felicia V S, Najla G, Ann P C, Madeleine M and Keneeth K C, “Inhibition of

Human Breast cancer cell proliferation and delay of mammary tumorigenisis by

flavonoids and citrus juices”, Nutrition and Cancer; 1996, 26: 167-181.

6) Catherine C, Malc S, Esther H L, Vadimir A, Krutorslaikh, et al. “Lack of tumour-

Promoting effects of flavonoids: studies on rat liver preneostatic foci and on In vivo

and In vitro gap junctional inter cellular communication”, Nutrition and Cancer;

1996, 26: 251-263.

7) Paul P, Ritra J, Ritva S, Mackku H, Lyly T, Eero P and Arpo A, “Dietary

flavonoids and the risk of lung cancer and other malignant neoplasms”, American

Journal of Epidemiology; 1997, 146: 223-230.

8) Fritz B, Tobias S, Albrecht K, Chaelotte B, Kent C, et al. “Selected novel flavones

inhibit the DNA binding or the DNA religation step of eukaryotic topoisomerase-I”,

Journal of Biological Chemistry; 1996, 271: 2262-2270.

9) Navarro-Núñez et al. "Apigenin Inhibits Platelet Adhesion and Thrombus

Formation and Synergizes with Aspirin in the Suppression of the Arachidonic Acid

Pathway", Jounal of Agricultural and Food Chemisry; 2008, 56 (9): 2970–2976.

10) The Wealth of India, A Dictionary of Indian Raw Materials and Industrial

Products-Raw Materials Series, Publication and Information Directorate, CSIR,

New Delhi, vol.2, 1950. p. 346-348.

11) Tabassum N and Ahmad F, “Role of natural herbs in the treatment of

hypertension”, Pharmacognosy Review; 2011, 5: 30-40.

12) Satoskar R S, Bandarkar S D, Nirmala N Rege, Pharmacology and

Pharmacotherapeutics, Popular Publications, 19th Edition, p.395-397.

13) Tripathi K D, Essentials of Medical Pharmacology, Jaypee Brothers Medical

Publishers, New Delhi 5th Edition, p.493-498.

14) Shafat A Quadri, Ariful N Qadri, Mark E, Hahn, Koren K Mann, David H, and

Sherr, “The Bioflavonoid Galangin Blocks Aryl Hydrocarbon Receptor Activation

and Polycyclic Aromatic Hydrocarbon”, Journal of Molecular Pharmacology;

2000, 58(3): 515-525.

15) Won Jun Lee, Joong-Youn Shim and Bao Ting Zhu, “Mechanisms for the

Inhibition of DNA Methyltransferases by Tea Catechins and Bioflavonoids”,

Journal of Molecular Pharmacology; 2005, 68(4): 1018-1030.

16) Sripal Reddy M, Raj Narayana K, Chaluvadi M R and Krishna D R,

“Bioflavonoids- Their pharmacokinetic interaction with cytochrome P450 isozymes

and P-glycoprotein”, Indian Journal of Pharmaceutical Sciences; 2001, 63(3): 187-

195.

17) Praveen Bhugra and Gulati O D, “Interaction of calcium channel blockers (CCB’s)

with Noradrenaline and Terbutalin on isolated right ventricle from normal and

diseased rats”, Indian Journal of Pharmacology; 1998, 30: 82-88.

18) Raj Narayana K, Sripal Reddy M, Chaluvadi M R and Krishna D R,

“Bioflavonoids Classification, Pharmacological, Biochemical Effects and

Therapeutic Potential”, Indian Journal of Pharmacology; 2001, 33: 02-16.

19) Sharma P, Kulshreshtha S, Sharma A L, Srivastava R K and Jain V K, “Cardiac

effects of calcium channel blockers in human volunteers”, Indian Journal of

Pharmacology; 1992, 24: 223-226.