Cardiovascular (CVS) agents

Introduction
CVS agents exert action on the heart or blood vessels in a direct or indirect manner
thereby affecting the distribution of blood to certain specified portions of the
circulatory system.

Autonomic CVS agents – CANNOTES

I.e. Almost every autonomic drug exerts marked cardiovascular activities

Non autonomic CVS agents

Eg- Digitalis and related derivatives / Parenteral Fluids/Diuetics
Classification of CVS agents
Classification based on ‘drug substances’
Autonomic agents
1. Sympathomimetics
Enhance BP
Lower reflexity
Stimulate heart
2. α−Adrenergic Blocking Drugs
Vasospastic conditions
Malignant Hypertension
Toxemic hypertensive crisis (preeclampsia)
3. β−Adrenergic Blocking Drugs
Essential hypertension
Portal hypertension
Angina pectoris
Instances of dysrhythmias

4. Anticholinesterase
Ensuring diagnosis of paroxysmal atrial
tachycardia and Myasthenia Gravis
5. Atropine and other Antimuscarinic Drugs.
Blocks cardiac vagus nerve in Adams Stokes Syndrome
some other bradycardias

Non autonomic agents

6. Digitalis and related derivatives
Peripheral and coronary dilators
Anti-dysrhithmic agents

7. Parenteral fluids (Saline with glucose, amino acids, electrolytes,

vitamins, and medications)
Management of severe shock

8. Diuretics
Adjunct in management of heart failure
Hypertension
Classification of CVS agents based on function
1. Cardiac Glycosides (Digitalis)
2. Antihypertensive and Hypotensive
3.Drugs Antiarrhythmic Agents
4. Vasopressor Drugs

Cardiac glycosides (Digitalis)

•Family of compounds derived from the foxglove plant (Digitalis spp

,belonging to Scrophulariaceae family) ,Hence the word digitalis

• Act on the heart by causing atrio ventricular conduction and vagal

tone.
• Positive Inotrope - increases force of contraction
• Digoxin and digitoxin are the two cardiac glycosides most frequently
used and have replaced other digitalis in cardiac therapy
Digitalis are Ionotrophic on heart

Atrioventricular conduction
(Treating atrial fibrillation )
Slowdown heart rate /
enhance the force of contraction
Vagal tone
(Treating congestive heart failure)

Atrial Fibrillation -
Irregular heartbeat
(arrhythmia) that can lead
to blood clots, stroke, heart
failure
Ionotrophic drugs do not prolong the life –span of an individual
having CHF

Why?
……………………………………………………………………………..

Digoxin cause diuresis and relief of oedema

How?
……………………………………………………………………………..
Electron microscopy Suggests a Deep cleft :The binding sight

MOHRAZ and SMITH, 1984

Laursen et al, 2014

Cristal structure confirms the actual binding sight
General Structure Activity Relationship studies seek
•Synthesis/derivetization 1. How does a drug fit in to receptor
•Computer aided molecular modeling 2. What part of the molecule effectively binds and
•Conformational energy studies pharmocologycal activity (Pharmacophore)
3. What are the best fit structures/conformmations
You need to know , For Digitalis

•Structure of the drug target?

•Binding sight of the target?

•How do scientists study structural requirements

in the drug?
 Digoxigein Digitoxigenin
Digoxin Digitoxin
Oral absorption 40-75% 90-100%
Half life 39h 7.6 days
(Fast onset sorter (Slow onset longer action/max.cumulative
action) action
Protein binding Low (~ 30%) High (90%)
Time for peak effect 3-6 h 6-12 h
Source Leaves of Digitalis lanata Leaves of Digitalis purpurea
 Digoxin Digitoxin
Administration Oral Oral, IM or IV
Dose < 10yrs age Adults.
Rapid digitalization, 600 mcg followed by 200 to
1-1.5mg divided in to two 400mcg every 3-6h
each after 6-8 h.
Maintainance,
0.125 to 0.5mg on a day

Write a short essay on

Digoxin and Digitoxin -Mechanism of Action
Points to note-
-
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-
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Self assessment questions

1. Write notes on cardiac glycosides ?

2.How cardeac glycoside receptors are designed ?
3. What is the difference between digoxin a digitoxin ? Explain with
respect to structures and give any one of its mechanism of action?
16.4. Vassopressor drugs

 Agents that cause vasoconstriction leading to

increased systemic and/or pulmonary vascular
resistance (SVR, PVR) and Mean Arterial Pressure
(MAP)
 Vasodilators that cause vasodialation are chiefly
employed for angina pectoris, cerebral or peripheral
vascular disorders

 Many drugs belonging to other classes also have both

vasopressor and ionotrophic effects.
• Main categories of adrenergic receptors relevant to
vasopressor activity:
– Alpha adrenergic receptor
– Beta-1, Beta-2 adrenergic receptors
– Dopamine receptors
Receptor Physiology
Receptor Location Effect
Alpha-1 Adrenergic Vascular wall Vasoconstriction
Increase duration of
Heart contraction without
increased chronotropy
Beta Adrenergic Beta-1 Heart ↑Inotropy and chronotropy
Beta-2 Blood vessels Vasodilation
Dopamine Renal Vasodilation
Splanchnic
(mesenteric)
Coronary
Cerebral
Subtype Vasoconstriction
 Examples and clinical applications
Drug Condition Dose
Buphenine Pheripheral Initially 6 mg oral thrice
,Buphenine Vascular daily may be enhanced
hydroclioride , dissorders to 36 or 48 mg per day
Nylidrin Menieres disease in divided doses
Hydrochloride
Isoxuspurine, Cerebral and Ora 20mg 4 times per
Isoxsupurine pheripheral day or Iv as 100 mg in
Hydrochloride vascular diseases 500 mL of sodium
chloride solution
Prenylamine, Prophylactically in Initial 180mg per day in
Prenylamine treatment of 3 divided doses
Lactate angina pectoris
Synthesis of Isoxsupurine hydrochloride
Synthesis of Prenylamine lactate
Mechanism of Action

Buphenine
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Isoxsupurine
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-
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Prenylamine
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Self Assessment Questions
1. What is a vasopressor drug? Describe.
2. Write about Prenylamine, its synthesis and
mechanism of action.
Vasopressor drugs (vasodilators) are chiefly
those employed for angina pectoris, cerebral
or peripheral vascular disorders.
A few important members of vasopressor drugs
are: Buphenine, Isoxsupurine and
Prenylamine.
 Introduction to
Computer Aided drug design
 Integration of Chemoinformatics and Bioinformatics

Genomic Large Molecule Small Computational

Biology Targets Assays Molecules chemistry

High
Throughput
Bioinformatics Cheminformatics
Screening
In silico
 Much About Structure

• Structure Function
• Structure Mechanism
• Structure Origins/Evolution
• Structure-based Drug Design
 Biological Structure
Sequence
3D
MESDAMESETMESSRSMYN
AMEISWALTERYALLKINCAL structure Structural Scales
LMEWALLYIPREFERDREVIL
MYSELFIMACENTERDIRATV
ANDYINTENNESSEEILIKENM
RANDDYNAMICSRPADNAPRI
MASERADCALCYCLINNDRKI polymerase
NASEMRPCALTRACTINKAR
KICIPCDPKIQDENVSDETAVS
WILLWINITALL
SSBs
Complexes
helicase

Organism primase

Assemblies
Cell
Structures

System Dynamics
Cell
 Bottlenecks in CADD
 Structures determined by NMR, computation,
or X-ray crystallography are static snapshots
of highly dynamic molecular systems
 Biological processes (recognition, interaction,
chemistry) require molecular motions and are
time dependent.
 To comprehend and facilitate thinking about
the dynamic structure of molecules is crucial.
 What is Molecular Modeling?
• Molecular modelling is a collection of
(computer based) techniques for deriving,
representing and manipulating the structures and
reactions of molecules, and those properties
that are dependent on these three dimensional
structures.
Keep in mind !!
Caveat: Is the interpolation and
extrapolation reliable?
•Also computers don’t do biology, they do sums quickly
 Capabilities of CADD
Capabilities of Molecular Modeling at Ranbaxy 3
1
2 Pharmacophore
Structure-based Ligand-based Development
Crystal Structure Homology Modeling
analysis SAR, 2D- & Lead
3D-QSAR Identification
Hits from
Computational Analysis Database Searches
of Protein-Ligand Interactions

In-Sillico
Modification of ligands within the BBB,Solubility,Caco-2 Prioritization of Hits
active-site for better binding &Toxicity Predictions

Lead
Optimization

Lead Hopping
Target (structure) based Ligand (analog)
drug design based drug design

Receptor structure is known Receptor structure is not known

Mechanism is known Mechanism is known/ unknown
Ligands and their biological Ligands and their biological
activities are known/ unknown activities are known
3D Structure of the Complex

Experimental
Information: The active
site can be identified
based on the position of
the ligand in the crystal
structures of the
protein-ligand
complexes
If Active Site is not KNOWN?????
Drug and Target : Lock and Key ?
Most of the drugs “FIT” well to their targets
Some “Locks” are known but not all !!
Study of protein crystals give the details of the “lock”.
Knowing the “lock” structure, we can DESIGN some “keys”.

This is achieved by COMPUTER Algorithms

This is called “STRUCTURE BASED DRUG DESIGN”

Algorithms

“Lock” structure “Key”constructed

(from experiment) by computer
 Variations on the Lock and Key Model

1- Which structure of the lock should be targeted?

2- Is the binding pocket a good target?
3- Is structure-based design relevant for my receptor?
-Is the 3D structure reliable?
-Is the binding pocket static enough?
4- Which key fits best?
5- What are the prerequisite physicochemical
properties for the key for better binding?
Drugs derived from structure-based approaches

Capoten Captopril ACE Hypertension 1981 Bristol-

Myers
Squibb
Trusopt Dorzolamide Carbonic Glaucoma 1995 Merck
anhydrase

Viracept Nelfinavir HIV protease HIV/ AIDS 1999 Agouron

(Pfizer)
and Lilly
Tamiflu Oseltamivir Neuraminidase Influenza 1999 Gilead and
Roche

Gleevec Imatinib BCR- Abl Chronic 2001 Novartis

myelogenous
leukaemia
 Virtual screening

Structure based drug design

Cycle
 Structure Based drug Design
HN

Docking and structure

O
based virtual screening
Docking Linking
Building

O
HN
? HN HN

O
? O
H ? O
H
? H

O O O

? S
O
HN
O
O
HN

H H
? O O
O O

O
H O S
HN

N O
Pharmacophore based ligand H

database search O
 Building (de novo design)
Know the protein ligand interactions

1. Hydrogen bonding
2. Hydrophobic interactions
3. Cation -ᴫ interaction ...
2011
1) Define Interacting Sites
HB donor/acceptor regions, Hydrophobic domain,
Exclusion volumes
2) Select Sites
3) Satisfy Sites
4) Join Functional Groups
5) Refine Structure
Linking (Fragment based approaches)
Linking (Fragment based approaches)

A. Binding site comprising

three binding pockets
B. Crystallographic screening
or virtual screening locates
molecular fragments that
bind to one, two or all
three pockets
C. A lead compound is
designed by organizing all
three fragments around a
core template
D. Growing out of a single
fragment
Linking Contd..
Pharmacophore based search
Pharmacophore based ligand database
search
Define Pharmacophore O Ligand O
H H Design H
O O O O
H
O
H
O O

O O O
O
O

Small molecule
O O
O DB Search H H
H
 Identify the binding regions Evaluate their disposition in space
(Difine pharmacophore)

Search for molecules in the library of

ligands for similarity
 Pharmacophore based ligand database
search Cotd…

Tools
Virtual screening by docking
• The process of “docking” a ligand to a binding site mimics the
natural course of interaction of the ligand and its receptor via a
lowest energy pathway.

• Put a compound in the approximate area where binding occurs

and evaluate the following:

– Do the molecules bind to each other?

– If yes, how strong is the binding?
– How does the molecule (or) the protein-ligand complex look
like. (understand the intermolecular interactions)
– Quantify the extent of binding.

•The orientation that maximizes the interaction reveals the most

accurate structure of the complex.
•The first approximation is to allow the substrate to do a random
walk in the space around the protein to find the lowest energy.
Docking of Ligand to the Active site of
Protein
 Docking
Individual docking not very meaningful ?
compound libraries are docked against an active site, hence the word screening

Ligand Active site

Non-Ligands
Small molecule library /database
 Free Energy of Binding
• Dock ligand into pseudo-
intercalation site
– Manual, automatic, and flexible
ligand docking
• Energy minimize to
determine DG complex
• Determine DGligand
_=interaction energy of
ligand with surroundings
when explicitly solvated

DGbinding = DHinteraction - T DSconformation+ DGsolvent

 Scoring is vital !
Detailed calculations on all possibilities would be very
expensive
The major challenge in structure based drug design to
identify the best position and orientation of the
ligand in the binding site of the target.
This is done by scoring or ranking of the various
possibilities, which are based on empirical
parameters, knowledge based on using rigorous
calculations
 Algorithms used while docking

• Fast shape matching (e.g., DOCK and Eudock),

• Incremental construction (e.g., FlexX, Hammerhead,
and SLIDE),
• Tabu search (e.g., PRO_LEADS and SFDock),
• Genetic algorithms (e.g., GOLD, AutoDock, and
Gambler),
• Monte Carlo simulations (e.g., MCDock and QXP),
Different programs use different algorithms

• Affinity • FlexX
• AutoDock • Glide
• Autodock Vina • GOLD
• BioMedCAChe • Hammerhead
• CAChe for Medicinal
Chemists • PRO_LEADS
• DOCK • SLIDE
• DockVision • VRDD
 Rigid Docking
• Shape-complementarity method:
find binding mode(s) without any
steric clashes
• Only 6-degrees of freedom
(translations and rotations)
• Move ligand to binding site and
monitor the decrease in the
energy
• Only non-bonded terms remain
in the energy term
• Try to find a good steric match
between ligand and receptor
 Flexible Docking

• Dock flexible ligands into binding pocket of

rigid protein
• Binding site broken down into regions of
possible interactions
hydrophobic

H-bonds

binding site from X- parameterised

ray binding site
• Then dock the molecule into pocket by matching
up interactions with ligand

parameterised binding site docked ligand

• Uses “random” translation, rotation, and torsion,
and look for a better binding mode.
• Even though we have
considered the ligand
to be flexible, the
active site was kept
as a rigid structure.
• The side chains of
the protein in the
vicinity of the active
site should be
flexible, but
computationally
more expensive.
• Receptor Mapping

The volume of the binding cavity is felt

from the ligands which are active or
inactive. This receptor map is derived by
looking at the localized charges on the
active ligands and hence assigning the
active site.
 Receptor Map
Proposed for Opiate
R3
Narcotics
(Morphine, Codeine, Heroin, etc.)

R2 R1
7.5-8.5Å

Anionic site
6.5Å
Focus of charge
*
Cavity for part of piperidine ring

Flat surface for aromatic ring

Structure modeling (Structure vs. Sequence)

- Homology modeling
- Fold recognition/ Threading
The 3D structures are used to understand protein
function and to design new drugs
 Homology modeling

Predicting the tertiary structure of an unknown

protein using a known 3D structure of a
homologous protein(s) (i.e. same family).
Assumption that structure is more conserved
than sequence

Can be used in understanding function, activity,

specificity, etc.
 Key step in Homology Modeling

•Alignment
–Multiple possible alignments
•Build model
•Refine loops
–Database methods
–Random conformation
–Score: best using a real force field
•Refine sidechains
–Works best in core residues
Structure Prediction by Homology Modeling
Structural Databases
SeqFold,Profiles-3D, PSI-BLAST, BLAST & FASTA

Reference Proteins
C Matrix Matching

Conserved Regions Protein Sequence

Sequence Alignment

Coordinate Assignment

Predicted Conserved Regions

Loop Searching/generation

MODELER

Initial Model

Structure Analysis
Sidechain Rotamers
and/or MM/MD
WHAT IF, PROCHECK, PROSAII,..
Refined Model
 Typical Contributions of CADD to Drug
Discovery Projects
Suggestions of structures which, when retrieved from a
compound collection or synthesized, were found upon
testing to be active or inactive as predicted
Development of structure-activity relationships
Visualization of receptor models, pharmacophoric
models, molecular alignments, or data models
Reanalyzing available data to achieve new insights
Creative search of available structures to find new leads
Identification of preferred sites for structure elaboration
Development of models to improve drug transport,
specificity, safety or stability
Development of mechanistic insights
Use of leads in one area to derive new leads in a related
assay
Establishment of useful databases of project structures
and properties
Computation of physical or chemical properties to
correlate with activities
 Pharmacokinetics play an extremely
important role in drug development

ADMET

•Absorption
•Distribution
•Metabolism
•Excretion
•Toxicity