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Concepts of Pharmaceutical Science
Competition experiment.
0.5 nM radioligand L* is bound to 1 ml samples of the same membrane samples used in the above experiments in the presence or absence of varying amounts of either compound A or compound B. After establishing equilibrium, the following counts were bound to the membranes (after subtracting nonspecific binding):
Concentration A (nM)
CPM bound with A
Concentration B (nM)
CPM bound with B
0
73001
0.05
72100
66365
0.1
71221
60835
0.3
67908
45626
0.5
64890
36501
1
58401
24334
3
41715
10429
5
32445
6636
10
20858
3476
30
8588
1197
100
2808
363
300
961
121
a). Which compound, A or B, is more a more potent displacer of the radioligand? Justify your answer why you think so.
b). Plot the fractional binding on a semilog plot and estimate the IC50s for both A and B.
c). Plot the results on a logit-log plot (pseudo-Hill plot) and determine the IC50s for both compounds A and B. Use these values for the next step. How close are the computed values to your estimate in part b?
d). Use the Cheng-Prusoff equation and the value of Kd for ligand L determined by the Scatchard plot in question 2 to calculate the Ki values for compounds A and B.
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