Workshop Cellular Pharmacology: Pharmacology

1. Draw a saturation curve for a ligand binding to its receptor.

Explain what the Kd is and how you can read it off the curve
What does the Bmax tell you and how can you estimate it from a curve?

concentration of binding
ligand (nM) (cpm)
10.00 280
7.00 301
3.00 240
1.80 205
1.50 171
1.20 127
0.60 70
0.00 0

2. Two drugs have been tested for their properties to cause a cellular response.
See the data below.

concentration (M) cellular cellular

response response
drug A drug B
2 x 10-10 2
5 x 10-10 5
1 x 10-9 0 7
2 x 10-9 3 13
5 x 10-9 10 18
1 x 10-8 16 25
2 x 10-8 25 30
5 x 10-8 38 35
1 x 10-7 53 36
2 x 10-7 66
5 x 10-7 78
1 x 10-6 79

a) Using graph paper, plot a log concentration response curves for drug A and B
b) Estimate the EC50 values for both drugs
c) Which drug is more potent and which is more efficacious? Explain your answer
3. This is the result for a saturation binding experiment. Plot a saturation curve.
Estimate Kd and Bmax for the ligand.

concentration of binding
ligand (nM) (cpm)
10.00 100.
7.00 98.
5.00 95.
3.00 85.
1.80 70.
1.50 60.
1.20 48.
0.60 30.
0 0

4. Two drugs have been tested for their properties to cause a cellular response.
See the data below. Which one is the more potent drug? Which one is more
efficacious?

concentration (M) cellular cellular

response response
drug A drug B
1 x 10-9 0 0
2 x 10-9 4 0
5 x 10-9 8 0
1 x 10-8 13 5
2 x 10-8 17 9
5 x 10-8 24 14
1 x 10-7 34 19
2 x 10-7 39 25
5 x 10-7 40 33
1 x 10-6 38
5 x 10-6 39
a) Using graph paper, plot a log concentration response curves for both drugs
b) Estimate the EC50 values for the drugs

5. Dose response data was collected during the preclinical testing of four drugs for
the treatment of acute heart failure. Which drug studied was the most efficacious?

6. Of the four drugs shown, which is the most potent?

7. Recent evidence indicates that receptors for many types of G-protein coupled
receptors (GPRCs) have constituitive (basal) activity independent of ligand activation.
In such systems, specific ligands (inverse agonists) can bind to these receptors,
producing effects that are opposite to those of a conventional agonist. In the graph
below, which drug displays a dose-response relationship consistent with this type of
behavior?

8. Drugs with low efficacy bind to receptors but do not fully activate them. Such
"partial agonists" can act as either as a weak agonist (in the absence of a full
agonist), or as a competitive antagonist (if a full agonist is present). Which curve best
reflects the effect produced by this type of agonist when it is administered alone?
9. Plot representative concentration-response curves for EACH of the following:

i) Plot a curve for drug A, estimate the EC50

ii) Plot a curve for the same agonist when you added in a competitive,
reversible antagonist with a dose ratio of 5
What is the EC50 of the agonist in the presence of the antagonist?

Concentration of drug Response of drug A

(M)
1 x 10-8 0
2 x 10-8 0
5 x 10-8 12
1 x 10-7 24
2 x 10-7 39
5 x 10-7 51
1 x 10-6 72
2 x 10-6 100
5 x 10-6 120
1 x 10-5 119

10. What type of ligand have you got here?

changes in receptor activation

150
A
100 B
50 C
0 D

-50
-100
-150
-200
-10 -9 -8 -7 -6 -5
ligand concentration log [M]
11. This graph illustrates the dose-response relationship for the effect of the beta
agonist isoproterenol on an isolated perfused heart, both alone and in the presence
of different fixed concentrations of Drug X. Based upon the data shown, Drug X is
most likely a(n):

12 This graph shows the concentration-dependent effects of norepinephrine on

arterial blood pressure, both alone, and in the presence of a fixed concentration of
Drug X. Which type of antagonist is Drug X?

13) Give a definition for the pA2 value and the dose ratio
14) In an experiment, an antagonist is added to drug A above at a concentration of
200nM. The dose ratio observed in this experiment is 2. What is the pA2 value of the
antagonist?

15) In further experiments, different antagonists are used which show different pA2
values.

Antagonist pA2 value

Beta 6
Delta 3
Epsilon 26
Gamma 24

Which of these is the most potent antagonist? Briefly explain your answer?

16) Below is a table with different examples of Kd, which drug has the highest affinity
for the receptor? Explain your answer

Drug Kd (nM)
A 3
B 2.5
C 30
D 45
E 60