Section 1 Pharmacology

Pharmacodynamics
Pharmacodynamic principles
 The drug-receptor interaction model describes a receptor as being in a state of equilibrium between its inactive form (R i)
and active (Ra) form. Receptor agonists change this equilibrium when they
bind and create a receptor-drug (R-D) complex.
o Full agonists have a high affinity for the receptor when in its
active state only
o Partial agonists have affinity for the receptor in both its active
and inactive state, but always greater affinity for Ra than Ri
o (Conventional) Antagonists have equal affinity for the receptor
in its active and inactive states, so that they encourage the receptor
to act exactly as it would in the absence of any bound ligand
o Inverse agonists have a greater affinity for the receptor in an
inactive state, resulting in the receptor spending even more time
than it otherwise would in its inactive state (e.g. benzodiazepines
acting allosterically on GABAA receptors to reduce Cl- flow
through this receptor complex)
 Drugs can either directly or indirectly bring about an effect. Direct effects
include activation of an ion channel or binding to an enzyme and altering
its activity. Indirect effects include the initiation of 2nd-messenger
signalling
 Drugs can be agonists, competitive inhibitors, allosteric activators or
allosteric inhibitors.
 Drug binding can be through covalent, electrostatic or hydrophobic
bonds.
o Covalent bonds (e.g. aspirin to platelet enzyme) are very strong
and usually irreversible,
o Electrostatic bonds can be of variable strength but are usually quite weak and readily reversible.
 Inert binding sites: drugs can also bind to molecules which do not have regulatory functions (‘inert binding sites’). Affects:
1. Distribution of drug in the body
2. The amount of free drug in the circulation
o Example: Albumin
 Stereo-isomerism (chirality) is also important; often there are differences in potency, toxicity, duration of action etc.
o Example: Ketamine: (+) enantiomer is more potent and less toxic than the (-) enantiomer

Receptors
 Receptors are key to the effect of drugs on the body because they:
1. Determine dose-response relationship
2. Result in selectivity of drug action
3. Mediate drug effects (both therapeutic and toxic)
 Many orphan receptors, whose binding ligand is current unknown, have been identified
 Common types of receptors are regulatory proteins (see below), enzymes (e.g. dihydrofolate reductase, the target of
antineoplastic methotrexate), transport proteins (e.g. Na/K ATPase, the target of digoxin) and structural proteins (e.g.
tubulin, the target of anti-inflammatory colchicine).

Dose-response curves
 In idealized (in vitro) systems, the relation between concentration and effect is described by a hyperbolic curve according to
the following equation:

E = effect at concentration C
Emax = maximal response that can be produced by the drug
EC50 = the concentration of drug that produces 50% of maximal effect

 This equation can be represented graphically as below, which closely resembles the mass action law:
  Receptor-effector coupling: The link between a receptor and the actual pharmacological response
A - The agonist response in the absence of
antagonist.

B - The result of treatment with a low

concentration of antagonist, where the curve is
shifted to the right but maximal responsiveness
is reserved because remaining available
receptors are in excess of the number required

C - A higher concentration of antagonist, at the

exact point where there are no more ‘spare’
receptors

D and E - Progressively higher antagonist

concentrations

Note the apparent EC50 in curves ‘D’ and ‘E’

approximates Kd (the binding affinity of the
agonist for the receptor), according to the Schild
equation.

 Spare receptors: Available receptors even when the agonist has elicited a maximal response.
o When spare receptors are available, adding a competitive antagonist (in the presence of an agonist) will have no
effect until all receptors, including those that were previously spare, are bound. T
o This phenomenon can be the result of
 A prolonged downstream effect that outlives the actual binding of agonist (temporal effects)
 An excess of the total number of receptors (quantal effects)

Types of antagonists
 Competitive antagonist
o With fixed presence of agonist and increasing dose of antagonist →
progressive inhibition of the agonist
 High antagonist concentrations prevent agonist response
completely
 High concentrations of agonist can completely surmount the effect
of a given concentration of antagonist → Emax remains the same
 Example: Naloxone vs. opiates
o Implications:
 Degree of inhibition by competitive antagonist depends on the
concentration of the agonist
 Clinical response to a competitive antagonist depends on the
concentration of agonist that is competing for binding to receptors
o The Schild equation can use this principle to determine the dissociation constant (Ki) for an antagonist:
[]

C’ = concentration of agonist required to produce effect

[I] = concentration of competitive antagonist
C = concentration of agonist normally required to produce effect (in the absence of
I)
Ki = dissociation constant for I
 Irreversible antagonist (or noncompetitive antagonist)
o Receptor antagonist that binds to the receptor in a functionally irreversible
fashion → Receptor unavailable to the agonist → Emax is decreased
 If there are ‘spare receptors’, Emax is decreased only at higher
concentrations of the antagonist!
  Example: Phenoxybenzamine, an irreversible α-adrenoreceptor antagonist used to control the
hypertension caused by catecholamines released by a pheochromocytoma
 Effect is irrespective of elimination of antagonist, and often can only be overcome by turnover of new
receptors
 Bad in overdose situations

 Partial agonist
o Produce a lower response at full receptor occupancy than do full
agonists → Emax is decreased
o Produce a concentration-effect curve that resembles that
observed with a full agonist in the presence of an antagonist that
irreversibly blocks some of the receptor sites
 Example: Pindolol (β-adrenoreceptor partial agonist),
compared to isoproterenol (full agonist)
 Chemical antagonist
o Antagonism by which one drug makes another drug unavailable
for receptor binding by rendering it chemically inactive
(Example: Protamine ionically binds heparin)
 Physiologic antagonist
o An antagonist effect that is mediated through an unrelated pathway (Example: Glucocorticoids increasing BSL in
opposition to insulin)

Signalling mechanisms
 Five basic mechanisms of transmembrane signalling are well understood:

1. Lipid-soluble ligand that crosses the membrane and acts on an intracellular receptor
o Example: NO → Stimulates intracellular guanylyl cyclase → ↑ cGMP
o Example: Glucocorticoids → Act on intracellular protein HSP90 → Relieves inhibition on transcription of specific
genes (this transcription takes some time, this in part explains the time lag between drug administration and clinical
effect; also explains why effect lasts after drug is gone)
2. Transmembrane receptor protein, whose intracellular enzymatic activity is allosterically regulated by a ligand that binds to
a site on the protein’s extracellular domain
3. Transmembrane receptor that binds and stimulates a protein tyrosine kinase
o Receptor is polypeptide with extracellular hormone-binding domain and intracellular enzyme domain (protein
tyrosine kinase, serine kinase or guanylyl cyclase).
1. Ligand binds to extracellular domain
2. Change in receptor conformation
3. Intracellular domain comes together → Becomes enzymatically active
4. Catalyses phosphorylation of tyrosine residues on different target signalling proteins → Modulates a
number of different biochemical processes with a single receptor and initiates a complex program of
cellular events.
o Examples: Insulin, PDGF and many other growth factors
4. Ligand-gated transmembrane ion channel that can be induced to open or close by the binding of a ligand
 Examples: Acetylcholine, serotonin, GABA, excitatory amino acids (Gly, Asp, Glu) → These are all
synaptic transmitters
 Work by increasing transmembrane conduction of a relevant ion → Altered electrical potential across the
membrane
 More specific example: ACh works on the nicotinic AChR → Opens Na+ channels → Na+ flows
down its concentration gradient into the cells → Depolarisation
 Time between binding of agonist and cellular response is usually milliseconds
 5. G-protein coupled receptor, a transmembrane receptor protein that stimulates a G- protein, which in turn modulates
production of an intracellular second messenger
o See below for details

G-proteins
 After the binding of an extracellular ligand to a membrane-bound receptor, there is a change in function intracellularly. The
intracellular mediator of this response is called a second messenger (see below). There are three things that occur to activate
the second messenger system:
1. Ligand binding to a specific receptor
2. Activation of G-protein (on intracellular membrane surface) in response to ligand binding
3. G-protein interaction with an effector element (usually an enzyme or ion channel), that changes the intracellular
concentration of a second messenger
 G-proteins
o These are proteins that, upon ligand binding, exchange GDP for GTP to
become active as a second messenger (G-GTP).
o After a delay, the GTP-protein complex autocatalyses the GTP to GDP
and is inactivated (G-GDP).
o Larger, heterotrimeric G-proteins (the first to be identified) are often
bound to receptors.
 Upon ligand binding, the α subunit dissociates from the βδ
subunit and both subunits (though mostly α) can act as second
messengers. GTP catalysis to GDP results in inactivation and
reassociation as a heterotrimer.
o G-proteins can act on phospholipase C, adenylyl cyclase, or directly on
ion channels.
o There are a number of different classes of G proteins.
o After a very brief (milliseconds) occupancy of a receptor by the ligand, the activity of the G protein may last for
tens of seconds → Amplification of the original signal (this also explains the phenomenon of ‘spare receptors’)

Second messenger systems

 cAMP
o Created by adenylyl cyclase acting on ATP.
 Adenylyl cyclase can be activated or inhibited by
heterotrimeric G-proteins after ligand binding. Gs α-
subunits will ↑ activity, Gi α-subunits will ↓ activity. The
response depends on which G-protein the receptor is
linked to.
o Activates protein kinase A (cAMP-dependent protein kinases),
which acts similarly to protein kinase C (on a different collection
of proteins)
 Activation involves cAMP binding to a regulatory dimer
(R2), releasing 2 catalytic chains.
 Involved in regulating many hormonal processes,
including:
 Release of stored energy (Example:
Isoprenaline, a β-agonist that stimulates
carbohydrate and fat metabolism)
 Conservation of water by the kidney (Example:
Vasopressin)
 Ca2+ homeostasis (Regulated by PTH)
 The rate and contraction force of the heart
(Example: Dobutamine, another β-
adrenomimetic catecholamine)
o Inactivated to 5’AMP by phosphodiesterase. Methylxanthines (Example: Theophylline, caffeine) inhibit
phosphodiesterases → ↑cAMP
o Cholera toxin prolongs Gs activity by preventing GTP autocatalysis. Pertussis toxin inhibits Gi activity. Both result
in increased intracellular cAMP.
 Calcium and phosphoinositides
o Normally, there is large differential between extracellular and intracellular calcium (Ca2+ECF is 12,000× Ca2+ICF).
o Ligand binding activates phospholipase C, either directly or via the Gq subtype of G-proteins, to catalyse
phosphatidylinositol (4,5)-disphosphate (PIP2) via this reaction:
 →
1. IP3 acts on ER to release Ca2+
2. DAG stays in the cell membrane and activates protein
kinase C, an enzyme that phosphorylates a number of
proteins (changing their conformation and activity)
o Calcium binds to many intracellular proteins, most notably
calmodulin
 Ca2+-calmodulin complex acts on Ca2+-dependent
protein kinases and other enzymes
o Removal of calcium from the cytosol is against its
electrochemical gradient. Through mechanisms:
 Ca2+ ATPase, an ATP dependent calcium transporter
 Na+, Ca2+ antiport
 SERCA pump back into ER
 cGMP
o Created by guanylyl cyclase, in response to NO.
o cGMP regulated ion channels and protein kinases have been
discovered.
 Example: Causes myosin light chain dephosphorylation
leading to smooth muscle vasodilation
o Broken down by phosphodiesterase

Efficacy vs. potency

 Potency describes the concentration of a drug required to give its full effect.
It is defined formally as the dose at which EC50 occurs. It depends upon the
affinity of the drug for the receptor to which it binds, and the efficiency with
which the drug-receptor interaction is coupled to a response.
o Potency of B > A
o Example: Fentanyl is much more potent than morphine
 Efficacy, in contrast, describes the strength of the maximal possible
response of a drug, irrespective of the dose required to produce this response.
o Efficacy of A > B
o Example: Buprenorphine is less efficacious than morphine (it is a
partial agonist)
 Shape of D-R curve is also important, as drugs with steep curves are likely
to have a narrow therapeutic index.
o Therapeutic index of C > D (most likely)

Quantal dose-effect curves

 A curve which shows the dose at which 50% of individuals exhibit a
specified quantal effect:
o ED50 (median effective dose): Dose of a drug at which 50% of
subjects exhibit a specific therapeutic effect.
o TD50 (median toxic dose): Dose of a drug at which 50% of
subjects exhibit the specified toxic effect.
o LD50 (median lethal dose): Dose of a drug at which 50% of
subjects exhibit the specified lethal effect.