SCREENING OF ANTI ANGINAL DRUGS

Presented by: SATISH. A. AKKI 1st M. Pharm Department of pharmacology SETs college of pharmacy Dharwad Subject: Pharmacological screening methods & clinical evaluation Subject incharge : Dr. PREETI.V.KULKARNI

CONTENTS
Introduction Heart

blood supply Ischaemic heart disease What is angina? Causes of angina Types of angina Symptoms o angina Pathophysiology of angina Tests of angina Preventive measures Anti anginal drugs Screening models Reference

Introduction
Angina pectoris was first described as a distinct clinical entity by WILLIAM HEBERDEN in the latter half of the 18th century

In the second half of the 19th century, it was found that amylnitrite could provide transient relief

Heart blood supply

Coronary

circulation (corona=crown) Two coronary arteries-right & left branch from ascending aorta Left coronary artery 1)anterior interventricular 2)circumflex branches Right coronary artery-1)posterior interventricular 2)marginal branches Coronary veins -coronary sinus -great cardiac vein -middle cardiac vein

Coronary arteries

Ischaemic heart disease

It

is defined as acute or chronic form of cardiac disability arising from imbalance between the myocardial supply and demand for oxygenated blood Narrowing or obstruction of coronary arteries Coronary artery disease Etiopathogenesis atherosclerosis -90% cases other causes -10% cases

Atherosclerosis

Effects of myocardial ischaemia

Asymptomatic

state Angina pectoris Acute myocardial infarction Chronic ischaemic heart disease Sudden cardiac death

What is angina?
Angina

(pain) or angina pectoris is the medical term used to describe the temporary chest discomfort that occurs when the heart is not getting enough blood Resulting from transient myocardial ischaemia It can no longer function at its full capacity Relieved by removing the stress or by taking sublingual nitro glycerin An episode of angina is not a heart attack Angina can be helpful warning sign

Causes of angina pectoris

Non-atherosclerotic coronary spasm arteritis embolism Atherosclerosis

Types of angina pectoris

Three types- based on difference in pathogenesis 1)Stable or typical angina 2)prinzmetals variant angina 3)Unstable or crescendo angina

Symptoms
Angina itself is a symptom (or set of symptoms), not a disease. Pain in the center of the chest Lightness Pain may spread to the shoulders, neck or arms Pain may be of any intensity from mild to severe Shortness of breath Anxiety or nervousness Sweaty skin It is always not easy to tell the difference between angina & heart attack

Symptoms

PATHOPHYSIOLSOGY OF ANGINA

Exams and tests

ECG Exercise

stress test Dobutamine echocardiogram stress test

Prevention
Stop

using nicotine in form Control high blood pressure Lower blood fats Control blood sugar Maintain healthy weight

ANTIANGINAL DRUGS
1. nitrates:Short acting:-glyceryl trinitrate. Long acting:-isosorbide dinitrate,isosorbide mononitrate,erytritil tetranitrate. 2. -blockers:- atenolol,metaprolol.propranolol

3.Ca-channel blockers:1.

2.
3.

Phenyl alkyl amines:-varapamil. Benzothiazepine:-diltiazem. Dihydropyridines:-nifedipine,amlodipine etc

4.K-channel openers:-nicorandil. 5.others:-dipyridamol,oxyphedrine.

SCREENING METHODS
Invitro models: 1.Isolated heart technique. 2.ca-antagonism in isolated rabbit aorta. 3.ca-antagonism in pitched rat. 4.Relaxation of bovine coronary aorta. 5.Coronary aorta ligation in isolated rat heart. 6.Isolated heart-lung prepration. 7.Plastic casts from coronary vasculature in dogs.

INVIVO MODELS
1.Occlusion of coronary artery. 2.Microspheres induced acute ischaemia. 3.Isoproterenol induced myocardial necrosis. 4.Stenosis induced coronary thrombosis model. 5.Electrical stimulation induced coronary thrombosis. 6.Models for coronary flow measurement. a.coronary in-flow measurement:-in anaesthetised dog. b.coronary out-flow measurement. i)electromagnetic flow meter ii)other techniques-like inert gas technique,radioactive technique.

IN VITRO METHODS
*Isolated heart / langendorff technique. - Principle ? - Procedure.

IN VIVO MODELS
Occlusion of coronary artery Principle Procedure: Dogs of either sex (30kg) Anesthetized with pentobarbitone sodium(35mg/kg,i.p) Trachea is cannulated Saphenous vein is cannulated for administration of test compound ECG is recorded continuously Femoral vein is cannulated & connected to pressure transducer Left ventricular pressure & heart rate are also measured using Millar microtip catheter inserted via left coronary artery

Heart is exposed through left thoracotomy Left anterior descending coronary artery is exposed & then ligated for 360min Test substance or vehicle is administered by i.v bolus infusion Hemodynamic parameters are monitored & at the end , animal are sacrificed Area at risk of infarction is measured using coronary arteriogram Left ventricle is cut into transverse section Slices incubated in p-nitro-blue-tetrazolium (0.25g/l) in order to visualize infarct tissue Blue stained is healthy, unstained is necrotic

Mortality, hemodynamic parameters & infarct size is determined Changes in parameters in treated animals are compared to vehicle control

Isoproterenol-induced myocardial necrosis

Principle Procedure: Wistar rats (150-200g) Pretreated with test drug or standard orally for at least a week Then, injected with 85mg/kg isoproterenol s.c on two consecutive days Mortality & symptoms are recorded in each group &compare to group injected with isoproterenol only After 48h of first dose of isoproterenol animals are sacrificed Heart is removed, weighed & preserved for histological evaluation

Before sacrificing the animal hemodynamic parameters can be recorded by cannulating the carotid artery Changes of parameters of drug treated animals are compared to isoproterenol controls

Stenosis-induced coronary thrombosis

Stenosis a Greek word-narrowing Procedure: Dogs(15-20kg) anesthetized with pentobarbitone sodium (30-40mg/kg i.p) Maintained on artificial respiration Heart is exposed at the 4th & 5th inter costal space An electromagnetic flow probe is placed on proximal part of left coronary artery to measure coronary blood flow Distal to flow meter, the vessel is clamped for 5sec A small plastic constrictor is placed around the artery at the site of damage

Constrictor is changed several times until required narrowing of the coronary artery is achieved In case the artery is occluded, the coronary artery is lifted to induce reflow Dogs with regular repeated cyclic flow variations of same intensity within a pretreatment phase of 60min are used for experimental purpose Hemodynamic parameters are recorded Test compound is administered i.v & the cyclic flow variations are registered for 2-5hr & compared to pretreated values In simple clamping of the coronary artery does not produce cyclic variations Cyclic flow variations are registered & compared to the drug treated group

Electrical stimulation-induced thrombosis

Electric stimulation can induce thrombosis in the coronary artery of pig Procedure: German landrace pigs(20-40kg) Anesthetized with ketamine(2mg/kg,i.m), metomidate(10mg/kg,i.p),& xylazine(1-2mg/kg,i.m) Maintained on artificial respiration Heart is exposed through left thoracotomy An electromagnetic flow meter is placed on the proximal part of the left coronary artery to measure coronary blood flow A vanadium steel electrode is placed in the vessel with the intimal lining & connected with teflon-coated wire of 9-volt battery, potentiometer & amperometer

 The intima is stimulated with 150A for 6hrs during which time an occluding thrombosis occurs Test drug is administered either s.c with electrical stimulation or 30 min after Hemodynamic parameters are measured by cannulating the femoral artery & connecting it to pressure transducer The time interval until the thrombotic occlusion of the vessel occurs & the thrombus size are determined At the end of experiment animals are sacrificed % change in mean values for occlusion time & thrombus size in drug treated group is compared to the control group Also changes in hemodynamic parameters is compared to pretreated values

References:
D. Tripathi, Essentials of Medical Pharmacology, 5th Edition, 2003, Jaypee Series, p. 437-40. H. P. Rang, M. M. Dale, J. M. Ritter, P. K. Moore, Pharmacology, 5th Edition, 2003, Churchill Livingstone, p. 494-98. Harsh Mohan, Textbook of pathology, 5th Edition, 2005, Jaypee Brothers Medical Publishers (P) ltd, p.316-18. H. Gerard Vogel and Wolfgang H. Vogel, Drug Discovery and Evaluation: Pharmacological Assays, 2nd Edition, 2002, Springer Publications, p. 595-643. S. K. Gupta, Drug Screening Methods, 1st Edition, 2004, Jaypee Series, p.
K.