PCOL 1 Drug Receptors 2017

Drug Targets
(including drug receptors)

A/Professor Ken Rodgers
School of Life Sciences
Copyright UTS, 2017 Slide 1 of 53

Lecture content
uTo describe the major types of target
proteins on which drugs act
uTo describe structure and activity of
receptors and ion channels
uTo describe the various forms of receptor-
effector linkages
uReceptor classification
uOther drug targets (enzymes and
transporters)
References
u Rang HP, Dale MM, Ritter JM

& Flower RJ, Henderson G
(2015) Pharmacology, 8th
Edition, Churchill
Livingstone, Sydney
u Chapter 3. How drugs act:

molecular aspects

Targets for drug action 1
uPaul Ehrlich drug action must be

explicable in terms of conventional
interactions between drugs and tissues

Targets for drug action 2
u Receptors
u 1. Ligand-gated ion channels (ionotropic receptors)
u 2. G-protein coupled receptors (metabotropic receptors)
u 3. Kinase-linked receptors
u 4. Nuclear receptors (intracellular receptors)
u Ion channels
u Voltage-gated ion channels
u Enzymes
u Transporters (carriers)
u Symports and antiports

Drug targets: receptors

Molecular pharmacology
Chemical messengers
Cellular
Types
Sensing
of receptor
element response
?
RECEPTOR
EFFECTOR LINKAGE
(Signal transduction
Copyright UTS, 2017 mechanisms) Slide 7 of 53
Receptor structure
uReceptors: An intracellular or cell-surface
protein with which a drug or a signalling
molecule interacts, to initiate a chain of
biochemical events in the cell or organism
uReceptors are proteins, glycoproteins or

lipoproteins these proteins provide a complex
3D shape
uFour receptor SUPERFAMILIES are known

Types of receptor-effector linkage

Ion channels
uGateways in the cell membrane
that allow the passage of particular
ions
Ligand-gated: trigger is agonist binding
Voltage-gated: trigger is a change in

transmembrane potential

Structure of ligand-gated ion channels
eg. Nicotinic acetylcholine receptor

Ionotropic receptors
uLigand-gated ion channel
u Location: membrane
u Effector: ion channel
u Coupling: direct
u Examples:
uNicotinic acetylcholine (ACh)
u-Aminobutyric acid (GABA)
uExcitatory amino acids (eg. NMDA, aspartate)
uGlycine
u Timescale: extremely rapid cell activation with a time

scale of milliseconds
u (note binding to receptor site is known as orthosteric
binding as opposed to allosteric binding)
uAllows ions to cross membrane thereby

causing:
umembrane polarisation (Cl-, K+)
umembrane depolarization (Ca2+, Na+)
umuscle contraction (nicotinic AChR of

skeletal muscle)
usignal transduction (via Ca2+ mobilisation or
influx)

uExample - Nicotinic AChR
u288 kDa heteropentamer with 5 subunits (, ,
, , , each 50 to 58 kDa) plus associated non-
selective cation channel (for +ve charged ions)
uEach subunit contains 4 membrane spanning
-helixes to form pore ~0.7 nm in diameter
u2 ACh molecules must bind to 2 subunits for
full activation conformational change to open
ion channel (twisting of subunits )
uAllows passage of Na+, K+, Ca2+ but at normal
resting potentials, Na+ is major ion

Nicotinic ACh receptor
20 membrane-
spanning alpha
helices
ACh binding
ACh binding
Structure of the nicotinic acetylcholine receptor (a typical ligand-gated ion channel).
side view (upper) and plan view (lower). The five receptor subunits (!2, ", #, $) form a clust
Nicotinic ACh receptor
- -
Na+ or K+
- -
Influx causes
- -
- -
depolarisation of
the cell
(sometimes Ca++)

Summary: ligand-gated ion channels
sometimes called ionotropic receptors

involved mainly in fast synaptic transmission
several structural families exist, the commonest
being heteromeric assemblies of four or five
subunits, with transmembrane helices arranged
around a central aqueous channel
ligand binding and channel opening occur on a
millisecond timescale.
examples: nicotinic acetylcholine, GABA type A
(GABAA) and 5-hydroxytryptamine type 3 (5-HT3)
receptors.

Other ligand-gated ion channels
Receptor heterogeneity within families gives rise to subfamilies of

different receptors in different tissues (eg nAChR in different
regions of the brain

The importance of G-proteins
The Nobel Prize in

Nobel Prize in
Physiology or
Medicine 1994 Chemistry for 2012
"for their discovery of G-proteins and the role of these proteins in signal
transduction in cells"
Robert J. Lefkowitz
Brian K. Kobilka
"for studies of G-protein

coupled receptors"
Alfred G. Gilman Martin Rodbell
USA USA
University of Texas, Southwestern National Institute of Environmental

Medical Center Health Sciences
Dallas, TX, USA Research Triangle Park, NC, USA
Copyright UTS, 2017 1941 - 1925 - 1998 Slide 20 of 53

Structure of G protein coupled receptors
eg. muscarinic acetylcholine receptor

G-protein coupled receptors
u G-Protein Coupled Receptors (GPCRs) or
metabotropic receptors
u Effector: channel or enzyme
u Coupling: G-protein
(affinity for guanyl

nucleotides (GDP/GTP)
u Examples:
uMuscarinic ACh
uAdrenoceptors
u Timescale: slow cell activation with a time scale of

seconds
Structure of muscarinic AcH
seven
transmembrane
helix (heptahelical)
structure
Family A: monoamine, neuropeptide and

chemokine receptors
Family B: calcitonin and glucagon receptors
Family C: glutamate and GABA receptors

2. G-protein coupled receptors 2
See: http://www.youtube.com/watch?v=V_0EcUr_txk

u Amplificationof signal:
one receptor can activate PRIMARY EFFECTOR
many G-proteins
u Active G-proteins can
cause effector enzymes to
produce many intracellular
second messengers
u Principal second
messengers:
u 1. Cyclic adenosine SECOND
monophosphate (cAMP) MESSENGER
u 2. Ca2+
u 3. phosphoinositides (eg
IP3 and DAG)

2. G-proteins: general scheme
Signal
AGONIST
(eg. NT, hormone)
Receptor
Primary Effector Transducer

G-PROTEIN
Second Messenger
Cellular
Secondary Effector
response
u G-proteins
u Involves intermediary G-proteins (Guanyl nucleotide-
binding protein) present in receptor-membrane complex
u G-proteins serve 3 roles
u 1.
G-proteins bind guanosine triphosphate (GTP) and GDP
G-proteins exist in 2 states:
u active form - GTP bound
u inactive form - GDP bound
u 2. G-proteins provide link between ligand-activated receptor

and effector (enzyme/ion channel)
u 3. G-Proteins have intrinsic GTPase activity which spontaneously
hydrolyses bound GTP to bound GDP (switch themselves off)

2. The function of G-proteins

2. G-proteins offer specificity
u Gi ('i' for inhibitory) and Gs ('s', for stimulatory) are
heterotrimeric signal transduction complexes ()
u -subunit interacts with a specific receptor (ie
muscarinic, dopamine, noradrenaline) and a specific
enzyme (eg. adenylate cyclase)
u On Gi / Gs activation, subunit can dissociate from


2. Primary effectors for GPCRs 1
u Targets for G-
proteins
u Adenylate cyclase/
cAMP system
u Phospholipase C/ Primary effector
inositol phosphate
system
u Regulation of ion
channels
Second messenger
cAMP is hydrolysed by
phosphodiesterases (PDE)
Cell specific (11 subtypes)
PDEs Can be inhibited for
example by caffeine
Protein
phosphorylation
AA = arachidonic acid
DAG = diacylglycerol,
Copyright UTS, 2017 IP3 = inositol triphosphate
Slide 31 of 53
Calcium vs. cAMP
Endoplasmic
reticulum
IP3
PKA
(protein kinase A)


Structure of kinase-linked receptors
eg. Insulin receptor

3. Kinase-linked receptors 1
u Effector: protein kinases
u Coupling: direct
u Examples:
u Insulin
u Growth factors eg.
Epidermal growth factor
(EGF), nerve growth
factor, platelet derived
growth factor (PDGF)
u Cytokine receptors eg.
interferon-gamma (IFN-)

uTimescale: cell activation with a time scale of
minutes to hours
uThese receptors consist of an extracellular
hormone binding domain and a cytoplasmic
enzyme domain
uEnzyme is usually a protein tyrosine kinase, but
can be a protein serine kinase, a protein
threonine kinase or guanyl cyclase (activation
of receptor by phosphorylation)

u Ligand binding
u Conformational change in receptor
causes inactive monomeric receptor
molecules to bind (noncovalently) to
one another to form active dimer
u This brings together intracellular
protein tyrosine kinase domains that
become enzymatically active
u Tyrosine (Y) residues in cytoplasmic
domains become phosphorylated (by
each other)
u Enzymatic activity is activated to
catalyse phosphorylation of substrate
proteins
u Cross phosphorylation intensifies or
prolongs allosteric action of hormone

Binding of
SH2 domains
of adaptor
proteins
Activation of
transcription
factors
<b>Transduction mechanisms of kinase-linked receptors.</b> The firnline Medical Textbooks Library for Students plus USMLE Steps 123 5/03/13 1:58 PM

3. Insulin receptor
u Insulin receptor is an
exception - it exists
as a dimer
u Receptor is
phosphorylated by
cytosolic kinases
u Phosphatidylinositol
3-hydroxy kinase
{PI(3)K}, makes
PIP2,PIP3
u Grb2, Sos, activates
Ras
u Activation of PI-PLC

3. Insulin receptor


Structure of nuclear receptors

4. Nuclear receptors 1
u Location: intracellular
u Effector: gene transcription
u Coupling: via DNA
u Examples:
u Sex steroids
u eg. Testosterone
u Glucocorticoids
u eg cortisol
u Mineralocorticoids
u eg aldosterone
u Hormones and vitamins
u eg. Vitamin D and thyroid
hormone

u Timescale: cell activation with a
time scale of hours
u Agonist-receptor complex acting
on DNA resulting in
u 1. transcription and translation of
mediator proteins or
u 2. repression of expression of
certain genes with inhibition of
production of specific proteins


4. Glucocorticoid receptor
Glucocorticoid
Eg. hydrocortisone Extracellular
Intracellular
Glucocorticoid Conformational change mRNA eg. lipocortin

receptor in receptor blocks PLA2 synthesis
- Anti-inflammatory action
DNA
Nucleus Glucocorticoid Gene

Copyright UTS, 2017
Responsive Element Slide 47 of 53
Receptor classification
u Drugs are designed to bind to specific targets and
these targets will only recognise certain drugs
u No drugs are completely specific - range of targets and
actions at those targets (basis of adverse reactions)
u Drug receptors are receptors for endogenous mediators
u Identification and Classification can be:

u Based on effect of selective antagonists or
representative agonists
u Differences in nucleotide sequence - functional
differences? (molecular biology methodology)
u Orphan receptors exist

Drug targets: enzymes
u Many drugs are competitive inhibitors of specific
enzymes eg captopril inhibits ACE
u Some are non-competitive inhibitors eg aspirin
inhibits COX
u Some drugs may be converted into abnormal
products eg fluorouracil (replaces uracil and
blocks DNA synthesis)
u In some cases enzymes in the liver can activate
drugs eg codeine is converted to morphine

Drug targets: enzymes

Drug targets: transporters
u A transport protein can transport molecules
across membranes (this may be required if they
are not very lipid soluble)
u Hydrolysis of ATP can provide the energy for
transport of substances against their
electrochemical gradient eg the sodium pump
u In some cases the transport of organic molecules
is coupled to the transport of ions (usually Na+),
either in the same direction (symport) or in the
opposite direction (antiport),

Drug targets: transporters

Examples of drug targets

PCOL 1 Drug Receptors 2017

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Drug Targets

(including drug receptors)

Copyright UTS, 2017 Slide 1 of 53

u Rang HP, Dale MM, Ritter JM

u Chapter 3. How drugs act:

Copyright UTS, 2017 Slide 3 of 53

uPaul Ehrlich drug action must be

Copyright UTS, 2017 Slide 4 of 53

u 4. Nuclear receptors (intracellular receptors)

Copyright UTS, 2017 Slide 5 of 53

Copyright UTS, 2017 Slide 6 of 53

uReceptors are proteins, glycoproteins or

uFour receptor SUPERFAMILIES are known

Copyright UTS, 2017 Slide 8 of 53

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Ligand-gated: trigger is agonist binding

Voltage-gated: trigger is a change in

Copyright UTS, 2017 Slide 10 of 53

eg. Nicotinic acetylcholine receptor

uExcitatory amino acids (eg. NMDA, aspartate)

u Timescale: extremely rapid cell activation with a time

uAllows ions to cross membrane thereby

umuscle contraction (nicotinic AChR of

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sometimes called ionotropic receptors

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Receptor heterogeneity within families gives rise to subfamilies of

Copyright UTS, 2017 Slide 19 of 53

The Nobel Prize in

"for studies of G-protein

Alfred G. Gilman Martin Rodbell

University of Texas, Southwestern National Institute of Environmental

Copyright UTS, 2017 1941 - 1925 - 1998 Slide 20 of 53

eg. muscarinic acetylcholine receptor

(affinity for guanyl

u Timescale: slow cell activation with a time scale of

Family A: monoamine, neuropeptide and

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Primary Effector Transducer

u 2. G-proteins provide link between ligand-activated receptor

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eg. Insulin receptor

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Glucocorticoid Conformational change mRNA eg. lipocortin

Nucleus Glucocorticoid Gene

u Identification and Classification can be: