PM1PCOL1

Reading School of Pharmacy

ANTAGONISTS

Receptor Theory 2: Antagonists

Copyright University of Reading LIMITLESS POTENTIAL | LIMITLESS OPPORTUNITIES | LIMITLESS IMPACT

Learning objectives
You will be able to
• Explain the differences between agonists and antagonists
• Define the types of antagonism, i.e. competitive and non-competitive
(allosteric or irreversible)
• Explain how concentration-response curves can be used to investigate modes
of antagonism and derive IC50 (antagonist potency)

2
Not all ligands are agonists…

Binding to Receptor 1 Concentration Response

Drug A is an agonist
of receptor 1

Drug B is an antagonist
of receptor 1

Drug A and B are ligands of Receptor 1 3

What is an antagonist?
• A drug that binds to a receptor but does not activate a response
• zero efficacy (Emax = 0)
• A drug that blocks the action of an agonist when bound

R + L LR LR*

4
Testing for antagonism
Agonist Concentration Response Antagonist Concentration Response
140

100 EC90 120

100 IC50: concentration

Response
Response

80 of antagonist to
50 60 reduce response by
40
50%
20
IC50
0
0
Cntl -10 -9 -8 -7 -6 -5 -4
-10 -8 -6 -4
log[Agonist]
Log [drug] (M) (M) Loglog [drug] (M) (M)
[Antagonist]

Select a concentration of agonist that Stimulate with the selected

given near maximal response (e.g. concentration of agonist and titrate
EC90) the antagonist against it
5
The IC50
• concentration of antagonist to inhibit 50% an agonist-induced response
• dependent on the assay conditions used
• IC50 can be referred to as antagonist potency

Which drug is
the most
potent
antagonist?

6
Modes of antagonism
Antagonist

Competitive Non-Competitive

Allosteric Irreversible

7
Competitive Antagonism
Agonist The higher the concentration of antagonist the
greater the chance they will bind unoccupied
Antagonist receptors before an agonist does

Reversible 8
ligand binding
Competitive
surmountable antagonism
R + A RA AR*
+ +
B B
A = agonist
B = antagonist
RB +A

9
Competitive Antagonism
• Competitive antagonists can be overcome by increasing the concentration
of agonist
• We say that the antagonist is ‘surmountable’

Agonist Concentration Response

• Competitive antagonists
decrease agonist 100 0 nM antagonist

potency (EC50) 3 nM Antagonist

Response

10 nM Antagonist
50 30 nM Antagonist
• Agonist efficacy (Emax) 100 nM Antagonist
remains unchanged
0
-10 -8 -6 -4 -2
log [Agonist] (M) 10
Not all antagonists are surmountable/competitive and/or reversible this is
termed insurmountable antagonism, two main types:

NON-COMPETITIVE
ANTAGONISM
IRREVERSIBLE AND ALLOSTERIC ANTAGONISTS
11
 Irreversible antagonists
Agonist If an antagonist binds a receptor irreversibly, it does
not compete with the agonist and receptors remain
Antagonist blocked even at high [agonist]

Reversible Irreversible 12
binding binding
Irreversible antagonists
R + A AR AR*
+ +
B B
A = agonist
B = antagonist
RB + A
non-competitive
13
Allosteric Antagonism
Agonist Allosteric antagonists bind to a different site (allosteric
site) than the agonist binding site (orthosteric site) and
Antagonist therefore bind non-competitively

Orthosteric
site

Allosteric site

Allosteric ligand 14
binding
Non competitive
allosteric antagonism
R + A AR AR*
+ +
B B
A = agonist
B = antagonist
RB + ARB
A
Non –competitive/
Allosteric antagonist 15
Non-Competitive Antagonism
(a.k.a insurmountable antagonism)

• Non-competitive antagonists cannot be overcome by increasing the

concentration of agonist
• We say that the antagonist is ‘insurmountable’

Non-Competitive Antagonism
• Agonist efficacy (Emax)
is reduced by non- 0 nM Antagonist
100
3 nM Antagonist
competitive antagonists EC50 10 nM Antagonist
Response

30 nM Antagonist
• EC50 is unchanged by 50 Emax 100 nM Antagonist

allosteric antagonists
0
• Irreversible antagonists -10 -8 -6 -4 -2
log [Agonist] (M)
effect EC50 and Emax 16
What do functional studies
(bioassays)
tell us about antagonists?
• Type of Antagonism
• Competitive/surmountable
• Non-competitive /insurmountable
• Allosteric?
• Irreversible?

Competitive
Competitive antagonism
antagonism
Agonist Concentration Response Non-competitive antagonism
Non-Competitive Antagonism

0 nM Antagonist
100 0 nM antagonist
100
3 nM Antagonist
3 nM Antagonist
Response

10 nM Antagonist
Response

10 nM Antagonist
30 nM Antagonist
50 30 nM Antagonist
50 100 nM Antagonist
100 nM Antagonist

0
0
-10 -8 -6 -4 -2
-10 -8 -6 -4 -2
log [Agonist] (M) log [Agonist] (M) 17
Other types of antagonism
• Agonists are ligands with efficacy

• Antagonists are ligands with zero efficacy

• Do some ligands have the opposite effect to agonists?

18
Constitutive activity
Agonist-induced receptor activation

R + A AR
AR*
Constitutive receptor activation

R R*

19
 Inverse agonists
Antagonist concentration responses in the
ABSENCE of agonist
100
90 Antagonist
80
Inverse Agonist If y-axis is ‘change in
70 response’ the graph will
Response

100
60 Constitutive 90 include negative values

Change in Response
80
70
50 activity 60
50
40 40
30
20
30 10
Inverse 0
-10
20 -20
agonism -30
10 -40
-14 -12 -10 -8 -6 -4 -2 0
0 log [drug] (M)
-14 -12 -10 -8 -6 -4 -2 0
20
log [drug] (M)
Constitutive activity
Constitutive receptor activation

Inverse agonism

21
Summary major types of agonists and
antagonists
120
100 Full Agonist
80
Response

60
Partial Agonist
40
20
0 Antagonist (neutral)
-20
Inverse Agonist
-40
-14 -12 -10 -8 -6 -4 -2 0
log [drug] (M)

Simple explanation of these types of ligand: 22

https://www.youtube.com/watch?v=_sM8KBZ9k4Q
Summary
• Antagonists are drugs that block the effect of agonists
• Antagonists have zero efficacy
• Competitive antagonists compete for the orthostatic binding site and are
surmountable
• Irreversible antagonists permanently block the receptor binding site
• Allosteric antagonists bind to a site distinct from the agonist binding site
• Inverse agonists are a type of antagonist that can inhibit constitutive receptor
activation (receptor activation in the absence of an agonist)

23
Questions – What kind of ligand is
this?

Saturation Binding Concentration Response Curve

150

100
Specific Binding

100

Response
50
50

0 0
-10 -8 -6 -4 -2 0 -10 -8 -6 -4 -2
log [drug] (M) log [Drug] (M)

24
Questions – What kind of ligand is
this?
Agonist Concentration Response

100 0 nM antagonist
3 nM Antagonist
Response

10 nM Antagonist
50 30 nM Antagonist
100 nM Antagonist

0
-10 -8 -6 -4 -2
log [Agonist] (M)

25
Questions – Which ligand has the
highest affinity?
150

Drug A
Specific Binding

100 Drug B

50

0
-10 -8 -6 -4 -2 0
log [drug] (M)
26
Questions – What is the IC50 of
this
antagonist?
Antagonist Concentration Response
140
120
• In log Molar (logIC50(M))?
100
Response

80 • In nano molar (nM)?

60
• In micro molar (μM)
40
20
0
Cntl -10 -9 -8 -7 -6 -5 -4
log [drug] (M) 27
Units
Numerical value Factor Prefix (symbol) Log 10
0.001 1x10-3 milli (m) -3
0.000001 1x10-6 micro (μ) -6
0.000000001 1x10-9 nano (n) -9
0.000000000001 1x10-12 pico (p) -12

28
Further study
• Rang and Dale’s Pharmacology (9th edition) Chapter 2
• Bear in mind this textbook goes beyond the level expected from this lecture
• Simples youtube video about types of ligand (
https://www.youtube.com/watch?v=_sM8KBZ9k4Q)

29