SUBJECT FORENSIC SCIENCE

Paper No. and Title PAPER No.9: Drugs of Abuse

Module No. and Title MODULE No. 23: Pharmacodynamics

Module Tag FSC_P9_M23

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 TABLE OF CONTENTS

1. Learning Outcomes

2. Introduction to Pharmacodynamics

3. Principle of Drug Action

4. Mechanism of Drug Action

5. The Transducer Mechanism

6. Summary

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 1. Learning Outcomes

After studying this module, you will be able to know about:

 Pharmacodynamics of Drugs
 Mechanism of Drug metabolism
 Identification of the receptor or enzyme which helps in drug action

2. Introduction to Pharmacodynamics

Pharmacodynamics is the study of biochemical and physiological effect of drug and

their mechanism of action at organ level as well as cellular level. Or, it may be
referred to as the association between drug concentration at the site of action and
subsequent effect, comprising the time and intensity of therapeutic and unfavorable
effects. The effect of a drug existing at the place of stroke is determined by the drug’s
binding with a receptor. Receptors may be present on neurons in the Central Nervous
System (e.g., Opiate receptors) to depress pain sensation, on cardiac muscle to affect
the intensity of contraction etc. For, most drugs, the concentration at the site of the
receptor determine the concentration of a drug’s effect.

Fig: Relationship of drug concentration to drug effect at the receptor site.

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 3. Principle of Drug Action

Drugs do not demonstrate latest functions on any system, organ or cell. It only alters
the pace of ongoing activity by different actions:

4. Mechanism of Drug Action

The Drug action depends on the biomolecules, typically a protein:

(1) Enzymes: All biological responses are taken out in catalytic influence of
enzymes. Drugs have capability to upsurge or reduce enzyme mediated reactions.
The enzyme stimulation is not common in drugs or generally by endogenous
substrates. Enzymes such as, Acetylcholine esterase, Choline Acetyltransferase,
cyclooxygenase, Xanthine oxidase, Angiotensin-converting enzymes, carbonic
anhydrase, HMG-CoA reductase, Dopa- decarboxylase, Monoamine oxidase,
Dihydrofolatereductase, DNA polymerase.

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 Enzyme inhibition- Common mode of drug action:
 Non-Specific Inhibition: Denaturation of proteins- strong acids, heavy
metals, alkalis, alcohol, phenol etc.
 Specific Inhibition: They are divided as per the effect of fluctuating
intensity of the enzyme's substrate on the inhibitor.

In competitive inhibition, the substrate and inhibitor will not bind to the enzyme at
the same time. This generally consequences from the inhibitor having an affinity for
the active site of an enzyme where the substrate also binds; the substrate and inhibitor
compete for access to the enzyme's active site.

This kind of inhibition can be resolved by appropriately high concentrations of

substrate (Vmax remains constant), i.e., by out-competing the inhibitor. Though, the
apparent Km (Michaelis Constant) will upsurge as it attains a higher concentration of
the substrate to reach the Km point, or half the Vmax.

Competitive inhibitors are generally same in organization to the actual substrate.

In mixed inhibition, the inhibitor can attach to the enzyme simultaneously as the
enzyme's substrate. Though, the binding of the inhibitor interrupts the binding of the
substrate, and vice versa. This kind of inhibition can be decreased, but cannot come
across by amplifying concentrations of substrate. Even though it is probable for
mixed-type inhibitors to join in the active site, this kind of inhibition usually falls out
from an allosteric effect where the inhibitor attach to various site on an enzyme.
Inhibitor binding to this allosteric site changes the conformation (i.e., tertiary structure
or three-dimensional shape) of the enzyme so that the affinity of the substrate for the
active site is reduced.

Non-competitive inhibition is a form of mixed inhibition where the binding of the

inhibitor to the enzyme reduces its activity but does not affect the binding of
substrate.

Due to this, the extent of inhibition depends only on the concentration of the inhibitor.
Vmax will decrease because the reaction proceeds efficiently, but Km will remain the
same as the actual binding of the substrate, by definition, will still function properly.

(2) Ion Channel

Proteins take part in Transmembrane Signaling and regulate ionic composition. Ions
like: receptor or voltage gated Na+, K+, Ca2+ and Cl- channels.

Drugs also target these channels:

 Ligand gate channels
 G-protein operated channels
 Direct action on channels

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 (3) Transporters

Drugs interact with these transport systems in which substrates gets translocated
through the membrane when they bind to specific transporters (carrier) - solute
Carrier Protein. They pump the metabolites or ions relative to the direction of
concentration gradient or against it.

Example: Probenecid (penicillin and uric acid), Furosmide (Na+, K+, Ca2+, Co
transport), Hemicholinium (choline uptake) and Vesamicol (Active transport to Ach
to vesicles).

(4) Receptors

It is described as a macromolecule or binding site situated on cell surface or within

the effector cell that act to distinguish the signal molecule/drug and initiates the
response to it, but itself has no other function, e.g. G-Protein Coupled Receptor. The
word receptor was introduced in 1909 by Paul Ehrlich. Drugs generally do not bind
directly with enzymes, channels, transporters or structural proteins, but act by
particular macromolecules.

John Langley has been credited with the theory of drug acting on receptors. While
studying the antagonistic effects of atropine against Pilocarpine- induced salivation,
he observed, ―There are several elements in the cells of gland and endings of nerve
cell with which both atropine and pilocarpine are competent of forming compounds‖.
Langley later stated these factors as ―receptive substance‖.

Fig: Diagram showing the receptor on the surface of the cell.

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 A receptor can exist in atleast two conformational states, active (Ra) and inactive (Ri).
If these conditions are in equilibrium and the stationary condition predominates in the
nonexistence of drug, then the basal signal results will be low. The extent from where
the equilibrium is fluctuating in the direction of the relative affinity of the drug for the
two conformations.

 Agonist: An agent that stimulates the receptor to create an effect analogous to

that of the physiological signal molecule, e.g. Muscarine and Nicotine.

 Antagonist: An agent that prevents the action of an agonist on a receptor or the

succeeding reaction, but does not have an effect of its own, e.g. atropine and
muscarine.

 Inverse agonist: An agent which turn on receptors to produce an effect in the

reverse direction to that of the agonist, example DMCM.

 Partial agonist: An agent which turns on a receptor to create sub-maximal effect

but antagonizes the turn on of a full agonist, example pentazocine.

 Ligand: Those molecules which attach selectively to specific receptors or sites

(only binding or affinity).

If the above mentioned terms are explained in terms of Affinity and Intrinsic
Activity, which means:

Affinity – It is a measure of propensity of a drug to bind receptor; the attractiveness

of drug and receptor

Intrinsic Activity (or Efficacy or IA) – It is defined as the property of a bound drug
to modify the receptor in such a way that creates an effect. Some drugs possess
affinity but not efficacy.

Then:

 Agonist : Affinity + IA (1)

 Antagonist : Affinity + IA (0)

 Partial agonist : Affinity + IA (0 - 1)

 Inverse agonist : Affinity + IA (0 to -1)

Drug- Receptor binding

D+R DR Complex
Affinity

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MODULE No. 23 : Pharmacodynamics
 DR Complex Effect (E)
 Covalent bonds are steady and fundamentally irreversible

 Electrostatic bonds may be strong or weak, but are typically reversible

Functioning of Receptor:

Two necessary roles of receptor are as follows:

 Recognition of specific Ligand molecule
 Transduction of signal into response

Fig: All domains are Ligand binding domain and then effectors domain undergoes functional
conformational change

Fig: Diagram showing Ligand binding domain

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MODULE No. 23 : Pharmacodynamics
  Cell surface receptors floats in cell membrane lipids.

 Roles are determined by the relation of lipophilic or hydrophilic sphere of the

peptide chain with the drug molecule.

 Non-polar hydrophobic piece of the amino acid stay hidden in membrane while
polar hydrophilic stay on cell surface.

 Hydrophilic drugs cannot cross the membrane and has to connect with the polar
hydrophilic piece of the peptide chain

 Binding of polar drugs in Ligand binding domain induces conformational

changes (alter distribution of charges and transmitted to coupling domain to be
pass on to effectors domain).

 Drugs have an effect on the physiological receptors and arbitrate reaction of

autacoids, hormones, transmitters and others such as cholinergic, adrenergic or
histaminergic etc.

 Drugs may proceed on true drug receptors - Benzodiazepine receptors.

5. Transducer mechanism
The majority trans-membrane signaling takes place by a minute quantity of diverse
molecular process (transducer mechanism). Large number of receptors shares these
handfuls of transducer mechanisms to produce a combined reaction. The four chief
categories are:
 GPCR ( G- Protein Coupled Receptor )
 Receptors with built-in ion channel
 Enzyme associated receptors
 Transcription factors (receptors for gene expression)

GPCR (G- Protein Coupled Receptor): Large family of cell membrane receptors
associated to the effectors enzyme/channel/carrier proteins by more than one GTP
turn on proteins (G-proteins). All receptors has common pattern of structural
organization. The particle has 7 α-helical membranes across hydrophobic amino acid
segments – 3 extra and 3 intracellular loops.

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MODULE No. 23 : Pharmacodynamics
 Fig: Diagram showing the GPC receptor

GPCR - 3 Major Pathways: There are three pathways-

1. Adenylcyclase: cAMP pathway

2. Phospholipase C: IP3-DAG pathway

3. Channel regulation:

 Stimulated G-proteins will on or off ion channels – Ca2+, Na+ or K+ etc.

 These effect may be present with no interference of any 2 messengers –

cAMP or IP/DAG

 Brings about depolarization, hyperpolarization or Ca2+ changes etc.

 Gs – Ca2+ present in medical drug proceed by impersonating or blocking the

actions of endogenous ligands that control the flow of ions by plasma
membrane channels in myocardium and skeletal muscles.

 Go and Gi – opens K+ channel of muscles and heart and closes Ca+ channel of
neurons.

 The natural ligands include acetylcholine, serotonin, aminobutyric acid

(GABA), and the excitatory amino acids (e.g. glycine, aspartate, and
glutamate).

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 Enzyme linked receptor: Two types of receptors:

1. Intrinsic enzyme associated receptors

 Protein kinase or guanylcyclase domain

2. JAK-STAT-kinase binding receptor

 Extracellular hormone has two-binding domain and a cytoplasmic enzyme

domain (mainly protein tyrosine kinase or serine kinase).

 On binding, the receptor changes from its inactive monomeric state to an

active dimeric state.

 Cytoplasmic area turns out to be phosphorylated on particular tyrosine

residues.

 Enzymatic activities are activated, catalyzing phosphorylation of substrate

proteins and then activated receptors catalyze phosphorylation of tyrosine
residues on different target signaling proteins, thus permitting a single
kind of turn on receptors to adapt a number of biochemical procedures.
Examples: Insulin - intake of glucose and amino acids and controls
metabolism of glycogen and triglycerides. Trastuzumab, antagonist of
such kind of receptor – used in breast cancer.

RECEPTOR REGULATION GENE EXPRESSION:

Lipid soluble biological indicator across the cell membrane and proceed on
intracellular receptors – NO works by exciting cGMP. Receptors for corticosteroids,
mineralocorticoids, thyroid hormones, sex hormones and Vitamin D etc. excite the
transcription of genes in the nucleus by binding with specific DNA sequence – called
- ―Responsive elements‖.

Hormones generate their effects after an interval period of 30 minutes to several

hours—the time needed for the production of latest proteins – gene active hormonal
drugs needs time become active (Bronchial asthma). A favorable or toxic effect
remains even after removal.

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics
 6. Summary

 The subdivision of Pharmacology that relates drug concentration to biological

effects in known as Pharmacodynamics.
 Fundamental principle of pharmacology is that drugs must interact with a
molecular target to exert an effect. Drug interaction with molecular targets is the
initiating event in a multistep process that ultimately alters tissue function.
 Agonists activate receptors to produce a response. Antagonists bind with
receptors but do not activate them or cause a response. They can essentially block
the activation of receptors.
 Partial agonists produce a response. However, this is less than would be expected
by a full agonistic drug.
 Inverse agonists are drugs which can reduce the normal activity of the cell.
 Competitive antagonists are drugs that prevent activation of the cell by their
normal agent.
 Non-competitive antagonists are drugs that may block the receptor but not in a
permanent way.

FORENSIC SCIENCE PAPER No. 9 : Drugs of Abuse

MODULE No. 23 : Pharmacodynamics