PHARMACOL

OGY PHL 313

Chapter 2
Drug receptors
  Receptors determine the quantitative relationship
between drug dose and pharmacologic effect
 Receptors are responsible for the selectivity of
drug action
What is  Receptors mediate the actions of pharmacological

Drug agonists and antagonists

Receptor
The existence of Receptors have Receptor proteins
receptors was first been isolated have been
identified from biochemically cloned and
observations of the sequenced.
chemical and
physiological
specificity of drug
effects
 The two theories explained the binding between
receptors and ligand are:
A. The lock and key theory (old
theory)
B. The two-state receptor model (new theory)

Receptor Lock and key:

theory • There is agonist and antagonist ligands
• Agonists have an affinity to the receptor and have
the efficacy to activate the receptor by doing
conformational changes
• Antagonists have an affinity to the receptor but
don't have the efficacy to activate the receptor by
doing the same conformational changes
 The two-state receptor model (new theory)
• Receptors (proteins, ion channels...etc) are present
in many different conformations at any point of
time they are called energy landscapes.
• One of the shapes is the most energetic favorable
Receptor conformation and most of the receptors will be in
that conformation. But still there is other less
theory energetic favorable confirmations present as well.
• Based on the amount of the receptors in the
conformational shape, the ligand action is
determined.
• Ligands are either full agonist, partial agonist,
antagonist, and inverse agonist.
What drugs target?
Drug targets transporters, enzymes, ion channels and receptors.
A) Regulatory proteins:
which mediate the actions of endogenous chemical signals such as neurotransmitters,
autacoids, and hormones. G-Protein Coupled Receptors (GPCR), Nuclear
hormones receptors
B) Enzymes:
may be inhibited (or, less commonly, activated) by binding a drug. Examples
include dihydrofolate reductase, the receptor for the antineoplastic drug
methotrexate; 3-hydroxy-3-methylglutaryl–coenzyme A (HMG-CoA) reductase,
the receptor for statins; and various protein and lipid kinases. Aspirin inhibits
the cyclooxygenase enzyme leading to increase in the substrate
(arachidonic acid) leading to a decrease in the prostaglandins PGH2
levels, and this is the wanted effect that induce the analgesic effect.
C) Transport proteins :
Na+/K+ ATPase is the cell surface receptor for cardiac glycosides such as
digitalis induce contractility. norepinephrine and serotonin transporter
proteins that are membrane receptors for antidepressant drugs

E) Other Structural proteins: extra/intra-cellular proteins inhibited (or

less commonly activated) by the binding of a drug. e.g. tubulin is the
receptor for colchicine (an anti-inflammatory agent)
Relation between drug • The relation between drug dose and the clinically
concentration and response observed response may be quite complex
• These curves represent the Michaelis-Menten
curves or rectangular hyperbolae (Biochemistry -
Enzyme kinetics)
• Emax: The maximum efficacy or effect of a drug.
Where the curve plateau and considered a measure
of the efficacy and should be measured on the Y
axis.
• EC50 or ED50: Effective concentration / dose that
gives half the maximal response. It is half the Emax
considered a measure of the potency and should be
plotted on the X axis.
Binding • It is a true and direct way to determine the affinity
to a receptor (Kd)
study • Bmax: The maximum number of receptor binding
sites, sites bound by drug at plateau
• The decrease in the Bmax means loss in binding
sites
• Equilibrium dissociation constant Kd : is the
concentration of the drug at which will occupy 50%
of the receptors at equilibrium.
• Equilibrium dissociation constant = drug
concentration with ½ max receptor occupancy
NOTE: Low Kd = High affinity
NOTE: Kd and EC50 may or may not be equal
B =(Bmax xC)/(C + Kd)
 Drug + Receptor -> Drug-receptor complex -> -> -> Effect
What happens here: -> -> -> ?
This is Coupling: What ties drug-binding to the end effect

• May be simple line: e.g. drug binds to a receptor -> opens

Coupling channel -> action potential (Effect).

The number of receptors bound is directly related to drug effect

Receptors • More often complex line: e.g. signaling cascades, many steps
involved like what we see in GPCR.

to Effects The effect increases disproportionately to receptor binding, the

pharmacological effect may long outlast drug-receptor complex
existence

“Coupling” is related to initial conformational change of a drug

binding a receptor:
• FULL agonists are more efficient couplers than
PARTIAL agonist
• Coupling efficiency is also impacted by the above
relationship (linear or pyramidal)
 Since the magnitude of the response of a “receptor” to a
drug must be proportional to the concentration of bound-
drug, it makes sense that:
1. Response and drug-binding curves are similar
2. EC50 is related to KD.

Relation This leads us to the concept of

between drug • Drug efficacy
• Drug potency

concentration
and response
 Drug efficacy: The maximum effect (Emax) that the drug
produce
Drug potency: A comparative measure refers to the different
doses of two drugs needed to produce the same effect

Drug Looking at the graph, we plot data as effect in the Y axis vs.
log of drug concentration [Drug] in the X axis. This makes it

Potency and easier to determine the EC50 and to compare drugs efficacy
and potency.

Efficacy
• Drug A and B have the same efficacy
• Drug A has greater potency than B or C, because the dose
needed to produce the same effect seen in drug A is
larger in Drug B and C.
• Drug C has lower efficacy than B
• Drug C is more potent than B at lower drug
concentrations
 Agonists may differ in how tightly they bind to their
receptors (potency) and in the effect they produce
(efficacy).

Some drugs may bind very tightly (are highly potent) but

Partial and produce only a modest effect (low efficacy).

Full agonists Therefore:

• Low efficacy drugs are termed (partial agonists)
• The structurally related drugs which produce a full effect
are called (full agonists)

• Partial agonists produce a lower effect at full receptor

occupancy than full agonists
• Not due to a difference in affinity; can more easily be
linked to coupling efficiency
 Partial agonists can competitively inhibit full agonist activity, How ?
 A is the endogenous ligand that produce certain effect.
 B usually is partial agonist as compared to A since they produce
the same effect
 But the sum of the two results in being B will lower the effect of

Partial and A in total (Blue dashed line)

Weak partial agonists are also competitive antagonists

Full agonists Clinically, drugs that increase heart rate as partial agonist are given
to people with tachycardia, because the total effect is lowering HR.
 • Chemical antagonist
One drug bind to another drug chemically, then it antagonizes and
inactivate its activity. E.g. Protamine inactivates Heparin so it will no
longer be able to dissolve blood clots

• Physiological antagonist

General One drug produce an effect that antagonizes the effect of another
drug physiologically. E.g. Insulin antagonizes the effect of

classes of
glucocorticoids hyperglycemia duo to adrenal cortex tumor or
glucocorticoids treatment resulting in lowering blood sugar levels.

antagonists • Pharmacological antagonist

One drug bind to a receptor in the body resulting in blocking the
endogenous agonist to bind to the same receptor and activate the
receptor pharmacologically. E.g. Propranolol blocks beta blockers in
the heart, so NEp will no longer be able to activate the receptor and
increase HR.
Pharmacological antagonists may be:
A. Competitive (reversibly displaced by agonists)
B. Noncompetitive (not reversibly displaced by agonists)
 Competes with the agonist at the same receptor, and reversibly
binds to receptors
• The Emax for the agonist remains the same for any fixed
concentration of antagonist
• The presence of antagonist will increase the ED50 of the
agonist,

Competitiv so the Drug-effect curve of agonist will shift to the

right.

e
antagonists
 Competes with the agonist at the same receptor, and irreversibly
binds to receptors
• The Emax for the agonist will be reduced duo to the permanent
damage of the receptors
• The antagonism can not be overcome by increasing the
concentration of the agonist

Non- • The presence of antagonist will will not effect the ED50 of the
agonist, so the Drug-effect curve will descend and the efficacy

competitiv will be lowered

e
antagonists
SIGNALING MECHANISMS & DRUG ACTION

How the drug (ligand) react with receptor to produce an

action

Most often, drugs act at a cellular

level
1. lipid soluble drug crosses
membrane, intracellular binding
2. drug binds trans-membrane
protein, catalyzes enzymatic
activity
3. drug binds trans-membrane
protein, signals phosphorylation
4. drug binds, conformational
change opens ion channel
5. drug binds, signals via second
messenger (GPCR) mechanism
Types of ligand reactions with
receptor
1. Lipid-soluble drugs: E.g. steroids (glucocorticoids, sex steroids, thyroid hormones)
Can cross cell membrane and act via intracellular receptors.
MOA: binding to DNA via response elements resulting in Gene expression changes (takes time), resulting in
transcription + translation.

2. Ligand-induced dimerization (Kinase receptor): E.g. trophic hormones; insulin, epidermal growth factor
receptor EGFR, transforming growth factor beta (TGFβ) receptors
Receptors are big transmembrane proteins with extracellular ligand binding domains and intracellular enzyme
domains
MOA: Ligand binds extracellular domain -> conformational change -> dimerization (homo- or hetero-) -> Kinase
domains together -> effect
Types of ligand reactions with
receptor
3. Ligand-induced phosphorylation: E.g. growth hormones, erythropoietin (EPO), interferons
Able to pass a signaling event from extra/intra-cellular. Similar to kinase receptor.
MOA: Ligand/Agonist binds to receptor -> dimerizes; activates JAK(s) -> R-P -> signals larger cascade through STAT-P -> STAT:STAT
dimerization -> dislocates from membrane -> travels to nucleus, and transcribes

4. Ion channels: E.g. Ca2+ channels. K+ channels

Simple coupling to effect. They are two types:
• Ligand-gated: L binds R, opens channel, ions flux down (electro) chemical gradients
• Natural ligands = NTs: ACh, GABA, 5-HT, Glutamate
• Ion channels are not all the same; multiple subtypes
• Voltage-gated Ion channels are similar in action, but not opened by agonist binding; triggered by changes
in membrane potential
Types of ligand reactions with
receptor
5. Second messenger system: E.g. G Protein coupled receptors
Most complicated of these coupling systems. Lead to great signal amplification
MOA: GPCR’s signaling in 3 parts:
1. Extracellular ligand binds to specific receptor
2. G-protein activated on cytoplasmic/intracellular surface
3. G-protein changes activity of some effector: Enzyme or Ion channel
Part of the ‘signal transduction’ pathways will increase intracellular second messenger like
• Cyclic AMP (cAMP)
• Phosphatidyl inositol (PI’s)
Types of ligand reactions with
receptor
List of the second messengers G-proteins, effectors and receptors
• cAMP
• Most famous/common intracellular 2nd messenger
• Mediates hormonal responses (e.g. moving stored energy from
fat cells), renal water conservation, calcium homeostasis, HR,
production of hormones, smooth muscle relaxation and many,
many more
• PI (hydrolysis)
• G-proteins stimulate the catabolism of membrane-bound PIP2
into DAG and IP3
• DAG stays at the membrane, activates kinases like PKC
• IP3 diffuses through the cytoplasm, binds calmodulin,
cascades further
• cGMP
• Only active in a few cell types, i.e. intestinal mucosa and
vascular smooth muscle
• Similar activity in those cells to cAMP
Receptor desensitization
Receptor desensitization
The decreased responsiveness that occurs with repeated or chronic exposure to agonist and is a general
feature of most signaling membrane receptors.

• Receptor-mediated responses to drugs and hormonal agonists often desensitize with time.
• After reaching an initial high response, the effect diminishes over seconds or minutes even in the
continued presence of the agonist.
• This desensitization is usually reversible.
• Thus, several minutes after removal of the agonist, a second exposure to agonist results in a similar
response.
In most instances, the molecular basis of desensitization is unknown. But one of the reason it will
be
caused by the delay in signaling cascade
A- Graded Dose-Response
Relations
• To choose among drugs and to determine appropriate
doses of a drug, the prescriber must know the relative
pharmacologic potency and maximal efficacy of the drugs in
relation to the desired therapeutic effect.
• These two important terms, often confusing to students
and clinicians.
Potency: refers to the concentration (EC50) or dose (ED50)
of
a drug required to produce 50% of that drug’s maximal
effect.

Maximal efficacy
This parameter reflects the limit of the dose-response relation
on the response axis. Drugs A, C, and D in Figure 2–15 have
equal maximal efficacy, and all have greater maximal efficacy
than drug B.
B- Shape of dose-response
curve
• While many drug dose-response curves approximate to the
shape of Michaelis-Menten relationships (e.g. A), some
clinical responses do not.
• Extremely steep dose-response curves (e.g. B) may have
important clinical consequences if the upper portion of the
curves represents and undesirable extent of response (e.g.
coma caused by a sedative-hypnotic).

• This steepness could result from cooperative interactions

of several different actions of a drug (e.g. on heart, brain
and peripheral vessels - all contributing to lower blood
pressure), they may also result from receptor effector
systems which require most receptors to be occupied
before a response is observed
C- Quantal Dose-Effect
Curves
• It is not always possible to construct graded dose-response
curves if the pharmacological response is quantal event
such as: prevention of convulsions, arrhythmia or death.
• One solution is to determine the quantity of drug required
to produce a specific magnitude of effect in a large number
of patients (or animals). The cumulative frequency
distribution of response is then plotted versus log dose.
 The quantal dese effect curve is characterized by the
median effective dose (ED50) - the dose at which 50% of
individuals show the specified quantal effect
 The dose required to produce a particular toxic effect in
50% of animals is called the median toxic dose (TD50).
 If the toxic effect is death of the animal, a median lethal
dose (LD50) may be defined.
Therapeutic
index
• Quantal dose effect curves permit an analysis of the
margin of safety (or selectivity in response) for a specific
drug.
• Therapeutic index (TI) is defined as the ratio of the dose
that produces the desired therapeutic (ED50) to the dose
that produces the toxic effect (TD50).
TI = TD50/ED50

• If TD50 = 500 mg and ED50 = 5 mg, the drug is 100-fold

more selective for the desired response and the
therapeutic index is 500/5 = 100, we call this wide TI
• If TD50 = 500 mg and ED50 = 250 mg, the drug is 2-fold
more selective for the desired response and the
therapeutic index is 500/250 = 2, we call this narrow TI

 The therapeutic index in humans is never known with

great precision.
 Clinically acceptable risk depends on the severity of
the
disease being treated
Variations in drug
responsiveness
Individuals may vary considerably in their responsiveness to a drug, the responses are:
1. hyporeactive -> reduced response
2. hyperreactive -> increased response
3. tolerance -> responsiveness decreases with continued drug administration
4. tachyphylaxis -> rapid development of tolerance
The mechanisms producing variation in drug responsiveness
A. Alteration in drug concentrations reaching a receptor: Patients may vary in the rate of absorption of a drug or
in clearing the drug from the circulation resulting in different responses
B. Variations in endogenous receptor-ligands concentration: propranolol markedly slows heart rate in patients
with elevated catecholamines. However, propranolol will be without effect on a well-trained marathon runner.
C. Alteration in receptor number or function: agonist can produce down-regulation of the receptor, while
antagonist can produce up-regulation of the receptors, which collectively results in different responses
D. Changes in components of response distal to the receptor When the diagnosis is correct and the drug is
appropriate, an unsatisfactory therapeutic response can often be traced to compensatory mechanisms in the
patient that respond to and oppose the beneficial effects of the drug. Compensatory increases in sympathetic
nervous tone and fluid retention by the kidney, for example, can contribute to tolerance to antihypertensive
effects of a vasodilator drug. In such cases, additional drugs may be required to achieve a useful therapeutic
result.
Clinical
Selectivity
No drug causes only a single specific effect. Each drug have various effects on the body due to:
A. beneficial and toxic effect that is mediated by the same receptor-effector mechanism: e.g. bleeding
caused by anti-coagulant therapy; hypoglycemic coma due to insulin. Toxicity may be avoided by careful
management of dosage.

B. beneficial and toxic effect that is mediated by the same receptor but in different tissues or by
different effector mechanisms: e.g. digitalis glycosides inhibit the Na-K ATPase. A beneficial effect is
augmentation of cardiac contractility. Toxicity effects include cardiac arrhythmias, Gl effects and
changes in vision (mediated by Na-K ATPase inhibition). The drug should be administered at lowest
dose that produces acceptable benefit. Also, anatomic selectivity in drug administration may assist.

C. beneficial and toxic effect that is mediated by different receptor-effector mechanism: e.g., Beta and
alpha-adrenoreceptor antagonists and agonists, the H1 and H2 antihistamines, nicotinic and muscarinic
blocking agents. All show selectivity for specific subclasses of receptor but nevertheless have
overlapping affinities for receptors within their category. The solution to increase the selectivity as
possible.
Homework
Questions
1. Explain what drugs target in the body?
2. Explain the relation between drug concentration and response?
3. What is the binding study, in brief, and what is the role of Kd?
4. What is the difference between efficacy and potency?
5. how can we distinguish between full and partial agonist?
6. What is the difference between competitive and non-competitive antagonists, explain with graph?
7. What are the types of antagonists? With examples
8. Name some clinical implication of competitive and non-competitive antagonists?
9. Name the five main methods used by a ligand to bind to a specific receptor and produce an action,
with
examples?
10. What is receptor desensitization?
11. What are narrow therapeutic index drugs and how they look like in a dose-response curve?
12. What is TD50 and LD50?
13. Why there are variations in drug responsiveness between people?