Receptors as Drug Targets

Compiled by:

Chikowe, I.

Basic Medical Sciences

College of Medicine
Malawi
1

Receptors

Receptors: are specific areas of certain proteins and

glycoproteins that are found either embedded in cellular
membranes or in the nuclei of living cells.

Cell surface/membrane receptor: receptor embedded in the cell

membrane and transfers chemical information from the extracellular
compartment to the intracellular compartment.
Nuclear receptor: receptor that exists in the intracellular compartment
and upon activation binds to regulator regions in the DNA and
modulates gene expression.

Ligand: any endogenous (messenger) or exogenous chemical

agent that binds to a receptor.
Binding domain: the general region on a receptor where a
ligand binds.
2

Receptors and Messengers

Receptors and their chemical messengers are crucial to the

communication systems of the body.
When the communication goes wrong, the body does not work
normally and this can lead to ailments like:

The problems could be 2 ways:

Depression
Heart problems
Schizophrenia
Muscle fatigue and many more.
Too many messengers being released leading to overheating of target
cells (metaphorically)
Too few messengers being released making the cell sluggish

3
So drugs can either increase messengers or block messengers.

Nerve

Nerve
Signal

Messenger
Receptor

Response

Nucleus

Cell

Cell

Examples of Chemical messengers

Simple molecular neurotransmitters:

Complex molecular chemical messengers:

Monoamines; e.g. acetylcholine, noradrenaline, dopamine, serotonin.

Amino acids; -aminobutylic acid (GABA), glutamic acid, glycine.
Calcium ion
Lipids; prostaglandins, purines (adenosine or ATP).
Neuropeptides; endorphins, enkephalins
Peptide hormones; angiotensin, bradykinin
Enzymes; thrombin.

Receptors are identified by specific neurotransmitter or

hormone that activates them: e.g. receptor activated by dopamine is
called dopaminergic receptor; cholinergic receptor for aceytlcholine;
adrenergic receptor or adrenoceptor for adrenaline or noradrenaline.

Nerve 1

Neurotransmitters do not undergo

reaction when they bind receptor.Blood
They leave receptor unchanged supply
after passing on their message. Hormone
Nerve 2
The binding of messenger induces
change in shape which causes the
opening of ion channel.
Neurotransmitters
A target cell may have various receptors specific to different
types of messengers.
Not all receptors activated by same chemical messenger are
exactly the same throughout the body. E.g.

adrenergic receptors in lungs slightly different from adrenergic

receptors in heart; due to variations in amino acid composition.

Cell surface receptor

Nuclear receptor

Structure and Function of

Receptors

Most receptors are proteins with various post-translational

modifications like covalent attachments of carbohydrate, lipid
and phosphate.

Responses to extracellular environment involve receptors that modulate

cellular components which generate, amplify, coordinate and terminate
post-receptor signaling via (cytoplasmic) second messengers. E.g. cyclic
adenosinemonophosphate (cAMP).

These secondary messengers promote a sequence of

biochemical events that result in an appropriate physiological
response
Signal transduction: the mechanism by which any message
carried by the ligand is translated through the receptor system
into a tissue response.
9

Examples of common bonding forms in drug receptor

interactions (minus van der waals forces)

10

Structure and FunctionMechanism

Receptors contain a binding site (hollow or cleft in the receptor

surface) that is recognised by the chemical messenger
Binding of the messenger involves intermolecular bonds
Binding results in an induced fit of the receptor protein
Change in receptor shape results in a domino effect
Domino effect is known as Signal Transduction, leading to a
chemical signal being received inside the cell
Chemical messenger does not enter the cell. It departs the
receptor unchanged and is not permanently bound

11

Illustration of Mechanism
Induced fit

Messenger

Messenger

Messenger

Cell
Membrane

Receptor

Receptor

Cell

Cell

Receptor
Cell
message
Message

Binding site

Binding site

Binding site:

A hydrophobic hollow or cleft on the

receptor surface - equivalent to
the active site of an enzyme
Accepts and binds a chemical messenger
Contains amino acids which bind the messenger

ENZYME

12

Receptor/Messenger Binding
M

M
M
RE

RE

Signal transduction

Binding site is nearly the correct shape for the messenger

Binding alters the shape of the receptor (induced fit)
Altered receptor shape leads to further effects - signal
transduction
Bonding forces:

Ionic, H-bonding, van der Waals.

13

Example
vdw
interaction

H-bond

Binding site

Ser

ionic
bond

Phe

CO2

Asp

Receptor

14

Induced fit - Binding site alters shape to maximize

intermolecular bonding
Phe

Phe

Ser

CO2

Asp

Intermolecular bonds not

optimum length for
maximum binding strength

Induced
Fit

Ser

H
CO2

Asp

Intermolecular bond
lengths optimised

15

Transmembrane signaling of
cell surface receptor

This is accomplished by only a few mechanisms:

Transmembrane ion channels: open or close upon
binding of a ligand or upon membrane depolarization
G-protein-coupled receptors: Transmembrane receptor
that stimulates a GTP-binding signal transducer protein (Gprotein) which then generates intracellular 2nd messenger
Nuclear receptors: Lipid soluble ligand that crosses the
cell membrane and acts on an intracellular receptor
Kinase-linked
receptors: Transmembrane receptor
proteins with intrinsic or associated kinase activity which is
allosterically regulated by a ligand that binds to the
receptors extracellular domain.
16

Summary of receptors

Ion Channels

Rapidly acting (milliseconds) transmembrane ion channels:

Multi-unit complexes with central aqueous channel. Upon
binding of a ligand, channel opening allows a specific ion travel
down its concentration gradient.

18

Control of Ion Channels

Receptor

Binding
site

Cell
membrane

Five glycoprotein subunits

traversing cell membrane

Messenger

Induced
fit

Cell
membrane

Gating
(ion channel
opens)

Cationic ion channels for K+, Na+, Ca2+ (e.g. nicotinic) =

excitatory.
19
Anionic ion channels for Cl- (e.g. GABAA) = inhibitory.

MESSENGER

ION
CHANNEL
(closed)

Cell
membrane
Cell

Ion
channel

RECEPTOR
BINDING
SITE

Lock
Gate

Ion
channel

ION
CHANNEL
(open)
Induced fit
and opening
of ion channel

Cell
membrane

Cell
membrane

Ion
channel

MESSENGER

Ion
channel

Cell
membrane

Cell

20

Voltage-gated ion channels:

Gating: controlled by membrane polarization/depolarization

Not controlled by binding of ligands, rather they sense the potential
difference across the cell membrane.
Selectivity: Na+, K+ or Ca+ ions
Important drug targets for local anaethetics

Intracellular ligand-gated channels:

Consist of 5 protein subunits with receptor binding site being present on

one or more of the subunits
Binding of neurotransmitter to ion channel receptor causes a
conformational change in protein subunits so that the second
transmembrane domain of each subunit rotates to open the channel
Ca+ controlled K+ channel
21
ATP-sensitive K+ channel.

Responsible for

neurotransmission
cardiac conduction
muscle contraction etc...

E.g: Cholinergic nicotinic

receptors is an example
to these type of
receptors.

22

G-Protein-coupled Receptors

GPCR: Large family of receptors with a probable

common evolutionary precursor. Transmembrane protein that
is serpentine in shape, crossing the lipid bilayer seven times.
The G-protein-coupled receptors are membrane-bound
proteins with 7 transmembrane
sections. The c-terminal chain lies
within the cell and the N-terminal
chain is extracellular.
They activate signal proteins called
G-proteins.
Location of binding sites differs
between different G-protein-coupled receptors.
23

Binding of messenger leads to opening of binding site for

signal protein. The latter binds and fragments, with one of the
subunits departing to activate a membrane-bound enzyme.

The rhodopsin-like family of G-protein-coupled receptors

includes many receptors that are targets for currently
24
important drugs.

Second messengers

Essential in conducting and amplifying signals from G-protein

coupled receptors.

cAMP

cGMP

Ca

DAG

IP3

25

Illustration of G-Protein-coupled Receptors Activation

messenger
induced
fit

closed

open

G-protein
split

Here, receptor binds messenger leading to induced fit; opens a binding site
for signal protein (G-protein) and the G-Protein is destabilised then split.
26

G-Protein-Enzyme-linked receptors

Spans the membrane once and may form dimers.

These receptors also have cytosolic enzyme activity as an
integral component of their structure.
Metabolism
important functions controlled by these receptors.
Growth
Differentiation
Most common Enzyme-linked receptors are:

EGF
PDGF
ANP
Insulin

tyrosine kinase activity

27

Illustration of enzyme linked receptor

In some cases: G-Protein subunit activates membrane bound

enzyme; binds to allosteric binding site; induced fit results in
opening of active site and Intracellular reaction catalysed.
Enzyme

Enzyme

active site
(closed)

active site
(open)

Intracellular
reaction
28

In other cases: Protein serves dual role - receptor plus

enzyme; receptor binds messenger leading to an induced fit;
protein changes shape and opens active site; reaction
catalysed within cell.
messenger
messenger
induced
fit

closed

active site
open

intracellular reaction

closed

29

Kinase-linked Receptors

Receptors directly linked to kinase enzymes.

Messengers binding leads to opening of kinase active site,
allowing a catalytic reaction to take place.
A good example of Kinase-linked receptors is tyrosine-linked
receptor:
Tyrosine kinase receptors have an extracellular binding site for
a chemical messenger and an intracellular enzymatic active site
which catalyzes the phosphorylation of tyrosine residues in
protein substrates. E.g. receptor for insulin and growth factor.

Insulin receptor is preformed heterotetrameric structure that acts as a

tyrosine kinase receptor.
Growth hormone receptor dimerises on binding its ligand, then binds
30
and activates tyrosine kinase enzymes from the cytoplasm.

31

Intracellular Receptors

Receptor is entirely intracellular.

Ligand must have sufficient lipid
solubility.
Primary targets of these ligandreceptor complexes are
transcription factors.

Steroid hormones exert their effects by

this receptor mechanism.

DNA

RNA

proteins

32

Illustration

33

Regulation of Receptors

Receptors not only initiate regulation of physiological and

biochemical function but are themselves subject to many
regulatory and homeostatic controls.
Controls include

regulation of synthesis and

degradation of the receptor by multiple mechanisms;

covalent modification,

association with other regulatory proteins, and/or

relocalization within the cell.

Modulating inputs may come from other receptors.

Receptors are always subject to feedback regulation by their
own signaling outputs.

Reduced responsivity: Chronic use of an agonist can result

in the receptor-effector system becoming less responsive
eg. alpha-adrenoceptor agents used as nasal decongestants
Myasthenia gravis: decrease in number of functional
acetylcholine nicotinic receptors at the neuromuscular junction.
Increased responsivity: Chronic disuse of a receptoreffector system can result in an increased responsiveness upon
re-exposure to an agonist.

Denervation super sensitivity at skeletal muscle acetylcholine nicotinic

receptors
Thyroid induced upregulation of cardiac beta-adrenoceptors
Prolonged use of many antagonists (pharmacological as well as
functional) can result in receptor upregulation.
35

Receptor upregulation

Most receptors are internalized and degraded or recycled with

age and use.
Antagonists slow use-dependent internalization
Inverse agonists stabilize the receptor in the inactive state to
prevent internalization.
The cell continues to produce receptors.

36

37

Drug Designing from

Ligand-Receptor
M

M
M
RE

RE

Signal transduction

Agonists: drugs designed to mimic the natural messenger

Agonists should bind and leave quickly - number of binding interactions

is important

Antagonists: drugs designed to block the natural messenger

Antagonists tend to have stronger and/or more binding interactions,

resulting in a different induced fit such that the receptor is not activated
38

Design of agonists

Agonists: bind reversibly to binding site and produce same

induced fit as the natural messenger - receptor is activated
Similar IMF bonds formed as with natural messenger
Agonists often similar in structure to the natural messenger

must have the correct binding groups

binding groups must be correctly positioned
must have the correct shape and size to fit the binding site
Agonist

Agonist

Agonist

Induced fit

RE

RE
Signal transduction

39

Design of agonist-Binding groups

Binding groups

van der Waals

binding region

H-bond
binding region
O

H
O2C

OH
NH2Me

Neurotransmitter

Ionic binding region

Binding site

Receptor

Know the structure of the natural chemical messenger and

identify the functional groups involved in the bonding with
receptor.

In the hypothetical neurotransmitter shown above, important binding

groups and respective interactions are: aromatic ring (van der waals),
alcohol (H-bonding), ammonium ion (ionic bonds).
40

O
NH2Me

O C
2

Receptor

O
NH2M e

Binding site

O C
2

INDUCED
FIT

Binding site
Receptor

Induced fit allows stronger binding

interactions

Design of Agonist
Ionic
binding
group

H-bonding
group

HO
van der Waals
-bonding
group

NH2Me
H

H2N

NH2Me

HO
NHMe

HO
H
H

H
Me

Hypothetical
neurotransmitter

NH2Me

Possible agonists with similar binding groups

Compare Binding groups:

Identify important binding interactions in natural messenger
Agonists are designed to have functional groups capable of
the same interactions
Usually require the same number of interactions
42

CH2Me

NH2Me

II

O
O

CH2Me

Binding site

Receptor

Structure I has one weak binding

group - negligible activity

NH2Me

Binding site

Receptor

Structure II has 2 of the 3 required

binding groups - weak activity

No interaction
OH
OH

O
N H 2M e

H
NH M e
2

O 2C

H
Binding site

2 Interactions only

Binding groups must be positioned such that they can interact

with complementary binding regions at the same time
Example has three binding groups, but only two can bind
simultaneously
Example will have poor activity.
44

Mirror
H

O
H

H
O

H
O

Binding site

3 interactions

M eH2N

NH2Me

Enantiomers of
a chiral
molecule

O
NH2M e

NH2Me

OH

Binding site

2 interactions

One enantiomer of a chiral drug normally binds more

effectively than the other
Different enantiomers likely to have different biological
properties.

45

H
Steric block
O
H

N
H2

CH 3
No Fit

H
O

Me
Me
Steric block

O2 C

Binding site

Agonist must have correct size and shape to fit binding site
Groups preventing access are called steric shields or steric
46
blocks.

Design of Antagonists

Antagonists bind to the binding site through IMF but fail to

produce the correct induced fit - receptor is not activated
Normal messenger is blocked from binding
Perfect Fit
(No change in shape)

Me
H

O C
2

Binding site

Level of antagonism depends on strength of antagonist binding and

47
conc. and increasing the messenger concentration reverses antagonism.

OH
O C
2

Receptor binding site

Extra binding regions

A binding site can have extra binding regions

48

Antagonists can form binding interactions with extra binding

regions neighboring the binding site for the natural messenger
Hydrophobic
binding region

Extra hydrophobic
binding region

HO

H-bond
binding region

HO
H
NH2Me

Hypothetical
neurotransmitter

Ionic binding
region

O
Asp

49

Induced fit resulting from binding of the normal messenger

Hydrophobic
region

Hydrophobic
region

HO

HO

HO

Induced fit

HO
H
NH2Me

H
NH2Me

Asp

O
Asp

50

Different induced fit resulting from extra binding interaction

Hydrophobic
region

Hydrophobic
region
Hydrophobic
region
HO

HO

HO

HO

HO
H

NHMe

NHMe

Asp

O
Asp

Initial binding

Different induced fit

51

Competitive (Reversible)
Antagonists
M
An
An
RE

Antagonist binds reversibly to the binding site

Intermolecular bonds involved in binding

Different induced fit means receptor is not activated

No reaction takes place on antagonist

Level of antagonism depends on strength of antagonist binding

and concentration

Messenger is blocked from the binding site

Increasing the messenger concentration reverses antagonism.

52

Irreversible Antagonists

Non-Competitive (Irreversible) Antagonists

X

Covalent Bond
X

OH

OH

Irreversible antagonism

Antagonist binds irreversibly (covalent) to the binding site

Messenger is blocked from the binding site

Different induced fit means that the receptor is not activated

Increasing messenger conc. does not reverse antagonism

Often used to label receptors.

53

O
O

HO

Cl

Cl

Cl
Cl

Propylbenzilylcholine mustard

Irreversible
binding

Cl Nu

Nu
Nu

Nu
Antagonist
binding site
Receptor

Agonist
binding site

Cl

Receptor

Non-competitive (Reversible) Allosteric

Antagonists
Binding site
unrecognisable

Binding site

ACTIVE SITE
(open)
Receptor
ENZYME

Allosteric
binding site

Induced
fit
(open)
Receptor
ENZYME

Antagonist

Antagonist binds reversibly to an allosteric binding site

IMF bonds formed between antagonist and binding site
Induced fit alters the shape of the receptor
Binding site is distorted and is not recognized by messenger

Increasing messenger concentration does not reverse antagonism.

55

Antagonists by the umbrella effect

Antagonist binds reversibly to a neighbouring binding site

IMF bonds formed between antagonist and binding site

Antagonist overlaps the messenger binding site

Messenger is blocked from the binding site

messenger

Binding site
for antagonist
Binding site
for messenger

Receptor

Antagonist

Receptor
56

Partial Agonist

Agents which act as agonists but produce a weaker effect.

Partial
agonist
H

H
O

1
H

NHM e

O 2C

Receptor

Slight shift
NHM e

O 2C

2
Partial opening
of an ion channel

O
H

Receptor

Possible explanations

Agent binds but does not produce ideal induced fit for maximum effect
Agent binds to binding site in two different modes, one where the agent
acts as an agonist and one where it acts as an antagonist
Agent binds as an agonist to one receptor subtype but as an antagonist
57
to another receptor subtype.

Inverse Agonists

Properties shared with antagonists

Bind to receptor binding sites with a different induced fit
from the normal messenger
Receptor is not activated
Normal messenger is blocked from binding to binding site
Properties not shared with antagonists
Block any inherent activity related to the receptor (e.g.
GABA receptor)
Inherent activity = level of activity present in the absence of
a chemical messenger
Receptors are in an equilibrium between constitutionally
58
active and inactive forms.

Explanation of how drugs affect receptor

equilibria
A) Resting state

Agonist binding site

Inactive conformations
B) Addition of agonist

C) Addition of antagonist

D) Addition of inverse agonist

E) Addition of partial agonist

Active conformation

Desensitization

Receptors become desensitized on long term exposure to

agonists
Prolonged binding of agonist leads to phosphorylation of receptor

Phosphorylated receptor changes shape and is inactivated

Dephosphorylation occurs once agonist departs

1
O2C

Ion channel
O2C (closed)

O
Agonist

Receptor

NH3

O
Agonist

O2C

NH3

Agonist

NH3
P

Receptor

Receptor

Induced fit alters protein shape Phosphorylation alter shape

Ion channel closes
Opens ion channel
60
Desensitization

Sensitization

Receptors become sensititized on long term exposure to

antagonists
Cell synthesises more receptors to compensate for blocked
receptors
Cells become more sensitive to natural messenger
Can result in tolerance and dependence
Increased doses of antagonist are required to achieve same
effect (tolerance)
Cells are supersensitive to normal neurotransmitter
Causes withdrawal symptoms when antagonist withdrawn
Leads to dependence
61

Sensitization

Neurotransmitter

Normal response

Antagonist

No response

Receptor
synthesis

No response
Receptor
synthesis
Sensitization

Response

Increase
antagonist

Excess response

Dependence

No response

Stop
antagonist

Tolerance

No response

Design of an antagonist for the

estrogen receptor
His 524
Me OH
H
H
Glu353
H

H
O

H2O

Hydrophic skeleton

Arg394
Oestradiol

Phenol and alcohol of estradiol are important binding groups

Binding site is spacious and hydrophobic
Phenol group of estradiol is positioned in narrow slot
Orientates rest of molecule
Acts as agonist

Action of the oestrogen receptor

Binding
site
AF-2
regions

H12

Coactivator

Coactivator

Oestradiol

DNA

Oestrogen
receptor

Dimerisation &
exposure of
AF-2 regions

Nuclear
transcription
factor

Transcription

Design of an antagonist for the

estrogen receptor
Asp351
N

Side
chain

His 524

Glu353

O
OH
H
O

Arg394
Raloxifene

Raloxifene is an antagonist (anticancer agent)

Phenol groups mimic phenol and alcohol of estradiol
Interaction with Asp-351 is important for antagonist activity
Side chain prevents receptor helix H12 folding over as lid
AF-2 binding region not revealed
Co-activator cannot bind

Tamoxifen as an antagonist
for the estrogen receptor
O
Me2N

CH2CH3

Anticancer agent