Mechanisms of Drug Action
Workbook 2
Radioligand binding, Biologics, Drug
Metabolism and Elimination
Student number:
Peer marker student number:
Deadline: 30th November 2018
Peer Mark /100
Final Mark /100
Please sign below if you agree that you do not wish to contest the mark you have
been given from the peer marking session
Signed ……………………………………………… Dated…………………………
Please give details of the questions where you are NOT satisfied with the mark
awarded.
1
Radioligand binding
The table below shows data obtained from a radioligand binding experiment.
1. Calculate the specific binding of the radioligand for this experiment and fill in the
blanks in the table. (5 marks)
Total binding Non-specific Specific binding
Concentration of binding
radioligand
(nM) (3H-ligand bound, (3H-ligand bound, (3H-ligand bound,
dpm) dpm) dpm)
0 0 0
0.25 12 0
0.5 41 1
1 122 3
2.5 142 6
5 193 15
7.5 219 31
10 248 42
15 276 68
20 304 89
25 311 102
Mark: /5
2
2. Plot the non-specific binding (NSB) and the specific binding versus radioligand
concentration on the graph paper below (plot it so there are two curves/lines on
the graph) (10 marks).
Mark: /10
3
3. Determine the Bmax and the KB for the radioligand in this experiment (5 marks).
Bmax = _____________ KB = ______________
4. Explain why the two curves/lines that you have plotted are different shapes (4
marks).
Interpretation of saturation data
5. The saturation binding experiment below shows binding of the same radioligand
to membranes obtained from different parts of the brain. Briefly explain what
these data show (4 marks).
60
H-ligand binding (units)
50
40
30
20
10
Specific binding
3
0
0 10 20 30 40 50 60
[3H-Ligand] (nM)
Mark: /13
4
6. In the experiment below two different radioligands that bind to different receptors
are used to do saturation binding on membranes from the same brain region.
Briefly describe what these data show (4 marks).
250
200
Specific3H-ligad
binding (units)
150
100
50
Specific binding
0
0 5 10 15 20 25 30
[3H-Ligand] (nM)
Mark: /4
5
Competition binding assays
The graph below shows the results from a competition binding assay, showing how 4
different displacing (competing) ligands affected the specific binding of a radioligand.
In this experiment the concentration of the radioligand used was 2 nM, and the K D of
the radioligand is 3 x 10-9 M.
% of specific binding in
100
absence of inhibitor
80
60 Displacer 1
Displacer 2
40 Displacer 3
Displacer 4
20
0
-12 -11 -10 -9 -8 -7 -6 -5 -4 -3 -2
log[Displacer](M)
7. Write down the order of affinity of these four displacing ligands for the receptor,
beginning with the ligand with the highest affinity (2 marks).
8. Write down the equation that is used to calculate the KD of the displacing ligand
from this type of experiment (2 marks).
Mark: /4
6
9. Calculate the KD values of the 4 displacing ligands at the receptor (8 marks).
Displacer 1
Displacer 2
Displacer 3
Displacer 4
10. Are these ligands agonists or antagonists at the receptor? Justify your answer
(2 marks).
Mark: /10
7
Biologics
Biologics (also called biopharmaceuticals) are pharmaceutical agents that are
produced by a biological process.
11. Monoclonal antibodies are one type of biologic. What are 5 ways in which
monoclonal antibody administration can produce a therapeutic effect (5 marks; 1
example is provided for you).
a. Deliver another substance (e.g. radioactive ligand) to the site of action
b.
c.
d.
e.
f.
12. For each of the types of biologic listed below, state briefly what the aim of
treatment with that type of biologic is (3 marks).
a. Polypeptides
b. Viral vectors
c. Oligonucleotides
Mark: /8
8
13. The table below contains the names of four antibody therapies that are either
licensed for use in humans, or are currently in clinical trials. Complete the table
detailing the type of antibody (i.e. fusion protein, chimeric, humanized or human),
the target that the antibody is designed to bind to, and one disease that the
biologic is aimed at treating (12 marks).
Antibody name Type of antibody Target Disease
Etanercept
Palivizumab
Solanezumab
Golimumab
14. List 2 advantages and 2 disadvantages that biologics have compared to small
molecule drugs (4 marks).
Advantages
Disadvantages
Mark: /16
9
Drug metabolism and Elimination
15. What are the differences between phase 1 and phase 2 reactions that take place
during drug metabolism (4 marks)?
16. Why do phase 1 reactions often precede phase 2 reactions (2 marks)?
The table below shows the plasma levels in a single patient (in µg/ml) of the
anticoagulant drug warfarin after a single oral dose under two conditions. In the first
condition, the patient was taking warfarin only. In the second condition, the patient
was also taking daily doses of the antibacterial drug rifampicin.
Warfarin plus
Time (hours) Warfarin alone
daily rifampicin
12 5.2 4.5
24 4.2 3
48 2.6 1.5
72 1.6 0.7
96 1
120 0.63
Mark: /6
10
17. Plot the data for the two conditions on the graph paper below (8 marks).
18. What is the half-life of warfarin in the two conditions (4 marks)?
Warfarin alone
Warfarin plus rifampicin
Mark: /12
11
19. Describe the process that may explain these results (4 marks).
20. Draw and label a graph in the space below to show the relationship between the
plasma drug concentration and time for a drug administered as a single i.v. bolus
dose. (Assume 1st order kinetics).
Show on the graph the likely effects on the relationship between plasma
concentration and time of:
a. Increase plasma clearance
b. Increased half-life for drug elimination
4 marks
Mark: /8
12
21. If the drug is eliminated solely by the mixed function oxidase system, show
diagrammatically the likely effects of the following on the plasma concentration
versus time relationship:
a. Renal disease (2 marks)
b. Liver disease (2 marks)
Mark: /4
13
