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DRUG ANTAGONISM
Competitive and Non-Competitive Antagonism

Antagonist: a drug that binds to the receptor without causing activation and therevy prevents the
agonist from binding.

Why are Antagonists so useful?

useful tools in research about natural hormone action and NT action
some key experiments in hormone understanding requires antagonists to identify receptors and
see which mediate different action in the body
used to identify things like enzymes or other molecules involved in signal transduction
pathways

MECHANISM OF ANTAGONIST ACTION

The action of an antagonist can be defined more generally into two different types:
mechanisms involving receptor (the receptor that agonist acts through)
Mechanisms involving receptor, more indirect.

Mechanism not involving the receptor

Physiological/ Functional Antagonism

The antagonist is not a blocker but is an agonist that works by producing an opposite biological effect
to the substance being antagonized. Each substance acts through its own receptors.

Example: isoprenaline dilates the airways

Beta2 agonist isoprenaline acts at 2 adrenoreceptor.
Muscarinic agonist- carbicol acts at mAChR
Isoprenaline dilates airways BUT carbicol antagonizes this doesn't affect beta receptor but
muscarinic receptor transduction mechanism inhibits this system that the beta receptor works
Beta receptor acts to raise cAMP whereas muscarininc receptor inhibitis adenylate cyclase,
reducing amount of cAMP in cells

Works in many aspects of physiology: consider parasympathetic and sympathetic system

which often counter each other
E.g. histamine causing bronchoconstriction
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Physiological antagonism

Dose Response Curve shows: The effect of carbachol at 2 concentrations, 1M and 10M, on the
relaxations of tracheal smooth muscle caused by isoprenaline. The responses are plotted as a
fraction of the maximum to isoprenaline.

Response: relaxation of airways by smooth muscle as concentration of isoprenaline is increased.

Adding 10 M carbachol shifts curve to the right familiar to competitive antagonists
observe similar kind of effect
Also observed with: Propranolol- non selective beta blocker put propranolol on this tissue,
the curve also moves to the right

Mechanisms involving the receptor

Competitive antagonism: binding of agonist and antagonist are mutually exclusive: if agonist is bound
to receptor, the antagonist cannot bind and vice versa. Only one or the other can be bound at any one
time.

Example: atropine competitively blocks the action of Ach on mAChRs

Noncompetitive Antagonism: agonist and antagonist can be bound at the same time, to different
regions of the the receptor molecule. (rarer)

Example: ketamine (an anaesthetic) reversibly blocks NMDA receptors in the brain by blocking the
NMDA receptor ion channel.

Widely used as horse anesthetic (dissociative get feeling of being high)

Blocks Ion channel of NMDA receptor separate to glutamate agonist binding site. Not
mutually exclusive
E.g. atropine and ach - ocompete for same binding site hence mutually exclusive
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REVERSIBLE COMPETITIVE ANTAGONISM

John Gaddum quantified competitive antagonism idea:

A: agonist, B: blocker, R: receptor

A + R AR with equilibrium constant KA allows us to derive Hill Langmuir equation to

describe occupancy of receptor by agonist

Can also derive it if it forms BR blocker binding to receptor, can calculate occupancy using
Hill-Langmuir equation

Gaddum Equation: accounts for agonist and blocker being present in the solution simultaneously.

This has the effect of making KA bigger agonist seems to have lower affinity higher
concentration of agonist needed to effect the same response
Gaddum equation is used to work out KB for the antagonist.
Using the antagonist on tissue only will show no visible change. But using agonist in the
presence of antagonist will produce different effects.

Note that increasing the agonist concentration overcomes the effect of a reversible antagonist
(ie. The block is surmountable).
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Increasing the antagonist concentration Dose-resposne curve shifts further to the right.
Will choose some response level for agonist e.g. 40% - what concentration of agonist is
needed to make that response alone/ what concentration is needed in presence of antagonist?
Amount you need to multiply agonist response by tot get initial response level is called dose
ratio, r
Assume: equal response to equal agonist occupancy.

DERIVATION OF SCHILD EQUATION

Assumption of equal responses in presence and absence of antagonist in absence of

antagonist, PAR is A/ KA + a
In presence of antagonist PAR = [A] * dose ratio / ka . Gaddum equation
Key assumption occupancy of receptor by agonist is same at theset wo points on curve. Can
make the Reponses equal look at equations, divide top and bottom line by r, r appears
underneath the term on slide.
That whole part under must equal 1, work out r.

[]
Schild Equation: = (1 + )

[]
1=

SCHILD PLOT: log( 1) = log[]

Schild plot take logs of either side of the equation

Gives straight line, where constant is log KB

Just need to know dose ratio
Identity of affinity of agonist (or conc) doesn't appear nothing about agonist in there.
We measure dose ratio from DR curve, we know concentration of agonist, we work out KB
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The Shcild plot

Example
Slope is close to 1
Intercept on x ais gives log of kb
E.g. 1.04 x 10^-9 molar
Antagonist atropine has kb of 10^-9 molar form muscarinic receptors
Response I these guinea pigs are involved in muscarinic receptor.
Useful as you study the antagonist and learn about agonist
ACH can also innervate ACH

Reversible competitive antagonism

The action of the antagonist can be overcome by a sufficient increase in the concnentration of
agonist (i.e. the antagonism is surmountable)
In the presence of the antagonist, the curve relating the log of the agonist concentration to the
size of the response is shift to the right in a parallel fashion.
The relationship between the magnitude of the shift (as expressed by the concentration
ratio) and the antagonist concentration obeys the Schild equation.
o Linear relationship between agonist dose ratio and antagonist concentration.

Practical Applications of Reversible Competitive Antagonism

The characterization and classification of receptors
The assessment of new competitive antagonists
The classification of agonists
o The dose-ratio is independent of the nature of the agonist 1 = []/
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Parallel shift with

Pyridylethylamine is less potent than histamine by 100-fold.
But the shift in the dose-response curve caused by antagonist is exactly the same- dose-response ratio
is independent of specific agonist.

New drugs on hormones

Developed propranolol and ciketidine
Rpopranalol is beta blocker, histamine or cimetidine is h2 antagonists
Mved from hormone 9higher )
By hving similar strucute found drugs which are competitive

Agonist KB
If antagonist is produced in same effect against different agonist, all agonists are working on
same receptor
Guinea pig atrium h2 response
Slope of schild plot = 1 = competitive action
Burnamide low affinity for H1, high affinity for h2 drug selectivity
Difference in kb points to fact that difernet receptors
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NON-COMPETITIVE ANTAGONISM

Binding of antagonist is independent of binding of agonist. They do not compete for a binding site.
Sometimes, when agonist is bound, it can disturb the protein structure. There will be a change
in protein conformation.
The change in conformation might change the affinity of the antagonist

If we suppose binding is independent, Kb for free receptor= Kb for agonist-bound receptor

KA for receptor = KA for antagonist bound receptor.
If KB or KA were not the same, might be an allosteric effect
Allosteric think about meaning about conformational changes propagated through protein
between agonist and agonist binding site, there is communication
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Reversible non-competitive antagonism

Dose response curve for agonist with increasing concentration of antagonist most top is
control.

Left: Non-competitive antagonist: Maximum of dose response curve is decreasing

Competitive antagonist, have parallel shift, for noncompetitive dose goes down

Important for exam questions draw sketches of dose response curve and point out these
characteristics

Complications
Right: noncompetitive antagonist- big receptor reserve (large proportion of spare receptors )
agonist can still produce maximum response even though a lot f receptors are blocked by
non-competitive antagonist
In most tissues, at first dose response curve moves parallel, when receptor reserve is blocked,
maximum decreases
Caused confusion with parallel shift of competitive antagonists- but it is not! \it is just acting on
a tissue with spare receptors
Plot this on schild plot, slope is STEEPER than one whereas comp antagonist have slope of
one.
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Distinction between agonist binding and receptor activation

The del Castillo and Katz mechanism

Katz encapsulated idea that the agonist can bind to the receptor.
After agonist is bound, there is a conformational change in the protein, which results in the
active receptor to give the response.
KA: binding.
E: Efficacy- how well the receptor activates upon binding

Now antagonist can compete with agonist binding to receptor or antagonist can interfere with E
so that receptor doesn't activate so well.

Non-Competitive Antagonism

If blocker binds to receptor, there can be agonist or blocker binding

After both agonist and blocker are bound receptor may or may not be able to activate.
o Value fo E may be different on each side
Or affinity of receptor may change (KA change)
Some blockers bind to active state of receptor (do this more preferentially).

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