Receptors

as Drug Targets

Medicinal Chemistry-III
(Pharm 4123):
Section A

Ref.: An Introduction to Medicinal Chemistry, 3rd ed. 2005, G.L.Patrick, Oxford University press
 Assignment

An research/review article on

Medicinal Chemistry
Desensitization & sensitization

Some drugs – bind strongly to receptor, switch it on, but

block the receptor after certain period of time.
Thus, they act as agonists, then antagonists.

Mechanism is unclear but possibly prolonged binding

of agonist to receptor results in phosphorylation of
hydroxyl or phenolic groups in receptor.

This causes the receptor to alter shape to an inactive

conformation in spite of occupied binding site.
Desensitization & sensitization

This altered tertiary structure is maintained as long as

binding site is occupied by agonist.

When drug leaves receptor is dephosphorylated and

returns to original resting shape.
 Desensitization & sensitization

On longer exposure to drug, receptor-drug complex – removed

completely from cell membrane by endocytosis.

Portion of membrane is ‘nipped out’, absorbed into cell and

metabolized.

Prolonged activation of receptor can result in reduction of

synthesis of receptor proteins.

So, best agonists bind swiftly to receptor, pass on their message

and leave quickly. But best angagonists tend to be slow to
add and slow to leave.
 Tolerance & dependence

Depriving a target receptor may induce synthesis of more

receptors. By doing so, cell gains a greater sensitivity that can
explain phenomena of tolerance & dependence.

Tolerance - a situation where higher levels of a drug are required

to get same biological response.

If a drug is acting to suppress the binding of a neurotransmitter,

then cell may respond by increasing number of receptors.
This would require increasing dose to regain same level of
antagonism.
Tolerance & dependence
 Tolerance & dependence
Neurotransmitter

Normal response

Antagonist

No response

Receptor Receptor Response

synthesis synthesis
Sensitization

Increase Tolerance
antagonist

Stop
Excess response antagonist No response

Dependence
 Tolerance & dependence

If drug is suddenly stopped, all receptors become available.

There is now excess of receptors which makes the cell
supersensitive to normal levels of neurotransmitter.

The situation is equivalent to drug overdose. Resulting

biological effects explain distressing withdrawal symptoms.

Withdrawal symptoms continue until number of receptors return

to original level. During this period, patient tempt to take
drug again to ‘return to normal’ and acquire dependence on
drug.
 Cytoplasmic receptors

Not all receptors – in cell membrane.

Some receptors – within the cell. So, ligands have to pass

through cell membrane (fatty in nature).

Example, steroid hormones cross cell membrane to reach

target receptor in cytoplasm (oestrogen receptor).
 Receptor types and subtypes

Receptors: identified by specific ligand or hormone which

activates them.

Dopaminergic receptor activated by dopamine

Cholinergic receptor activated by acetylcholine

Adrenergic receptor or adrenoceptor activated by adrenaline

 Receptor types and subtypes

Not all receptors activated by same ligand are same throughout

the body.

Adrenergic receptors in lungs are slightly different from those

in heart.

These differences arise from slight variations in amino acid

composition. If variations are in binding site, drugs can be
designed which can distinguish between them e.g.,
adrenergic drugs can be ‘lung’ or ‘heart’ selective.
 Receptor types and subtypes

There are various types & subtypes of a particular receptor

identified by numbers or letters.

For historic reasons, types of cholinergic receptors named

after natural products.
Receptor types and subtypes
Receptor types and subtypes

Current emphasis is to design drugs as selective as

possible for receptor types & subtypes so that drugs
are tissue selective and have less side effects.

Examples:

All antipsychotic agents antagonize D2 & D3. but

blockade of D2 leads to some side effects, so a
selective D3 antagonist may have better properties.

M2 agonists for heart irregularities

Β2 agonists for obesity.

 Affinity, efficacy and potency
Affinity: a measure of how strongly a drug binds to receptor.

Efficacy: a measure of maximum biological effect that a

drug can produce as a result of receptor binding.

Compound with high affinity does not necessarily have high

efficacy e.g., an antagonist can bind with high affinity but
has no efficacy.

Potency: amount of drug required to achieve a defined

biological effect.

The smaller the dose, the more potent the drug.

Affinity, efficacy and potency
Measurement of Affinity: using a process, radioligand
labelling.

Known ligand for receptor is labelled with radioactivity

Added to cells or tissue for binding to receptors

After reaching equilibrium, unbound ligands removed by

washing, filtration or centrifugation

Extend of binding measured by detecting amount of

radioactivity present in cells or tissue, & amount of
radioactivity that was removed.
Affinity, efficacy and potency

Equilibrium constant for bound vs unbound radioligand

is dissociation binding constant (Kd)

[L] & [LR] obtained by measuring radioactivity of

unbound ligand and bound ligand respectively.
Affinity, efficacy and potency

Total no. of receptors equals no. of receptors occupied by

ligand [LR] & those that are unoccupied [R]

No. of receptors unoccupied by ligand is

Substituting [R] into previous equation & rearranging

leads to Scatchard equation, where [LR] & [L] are
measurable:
 Affinity, efficacy and potency
Kd & Rtot can be determined by drawing a graph based on
number of experiments where different conc. of known
radioligand are used (line A).

Scatchard plot

Slope is a measure of radioligand’s affinity for receptor & kd.

 Affinity, efficacy and potency
To determine affinity of a novel drug not radioactively labelled,
repeat radioligand experiments in presence of unlabelled test
compound (displacer) which competes with radioligand for
receptor’s binding site; resulting in different line (X).

Scatchard plot
 Affinity, efficacy and potency
Agents that bind to receptors at an allostaric binding site do not
compete with radioligand for same binding site, resulting in
lower total number of available receptors (Y).

Scatchard plot
 Affinity, efficacy and potency
Data from previous experiments – to plot a graph comparing %
bound radioligand vs conc. of test comp.

Resulting in a sigmoidal curve termed displacement or inhibition

curve from which IC50 of test comp. can be identified.
Affinity, efficacy and potency

Inhibitory or affinity constant (Ki) for test compound is

same as IC50 value in case of non-competetive
interactions.

For competetive interactions, Kj depends on level of

radioligand:

Kd = dissociation constant for radioligand

[L]tot = conc. of radioligand used in experiment.
 Affinity, efficacy and potency
Efficacy: maximum possible effect resulting from receptor-ligand binding.

Potency: conc. of drug to produce 50% of maximum possible effect (EC50)

The smaller the EC50, the more potent the drug.

pD2 is taken as measure of potency where, pD2 = -log EC50

 Affinity, efficacy and potency
Schild analysis: to determine Kd of competetive antagonists

Schild plot: compares reciprocal of dose ratio vs log-conc. of

antagonist

Dose ratio: agonist conc. required to produce a level of effect

without antagonist compared to agonist conc. required to
produce same level of effect with antagonist.