PHC62004 Basic Pharmacology and Toxicology with Health

Informatics

Answer Sheet - Practical 2:

Computer Assisted Learning (CAL) -

Effects of Antagonists on Acetylcholine
[6% of total marks (30%) for Practical component]

Name of student (Student ID): Date: 10 May 2021

Christine Liew (0349225)
Ng Zhi Wei (0348618)
Choo Shi Ying (0344403)
Low Jing Yie (0344203)

Part I: Concepts of Pharmacodynamics - Antagonists

1. Describe pharmacokinetics antagonism. (3 marks)

Pharmacokinetics antagonism is the result of one drug affecting another drug to reduce their
effect by reducing its absorption, altering its distribution, or increasing its rate of
elimination.

2. Discuss the mode of action of functional antagonist and chemical antagonist. Include
an example for each type. (2, 1, 2 x 2 marks)

The mode of action of functional antagonist is it binds to a different receptor molecule,

producing an effect opposite to that produced by the drug it antagonizes. For example, the
antagonism of the bronchoconstrictor action of histamine by epinephrine’s bronchodilator
action. In contrast, the mode of action of chemical antagonist is it interacts directly with the
drug being antagonized to remove it or to prevent it from reaching its target. By example,
dimercaprol, a chelator of lead and some other toxic metals.

Page 1 of 3
3. State the differences between a competitive antagonist and an irreversible antagonist by
completing the table below. Some columns are filled as examples. (6 x 1 mark)

Type of antagonist
Difference
Competitive Irreversible
It changes the receptor to decrease the
Bind to the receptor site in a
efficacy of the agonist or irreversibly
Mode of binding reversible way without activating the
blocks the agonist from combining
effector system for that receptor.
with the receptor.

Effects on agonist Reduces potency of agonist Decreases the efficacy of the agonist.

The agonist if given in a high enough

Effects of concentration, can displace the Cannot be overcome by adding
antagonist antagonist and fully activate the more agonist
receptors.

The concentration-response curve Causes a downward shift of the

Changes in shifted to higher dose which maximum, with no shift of the curve
concentration- horizontally to the right on the on the dose axis unless spare
response curve dose axis, but the same maximal receptors are present, and the potency
effect is reached. is the same.

Part II: OBSim Simulation - Data Interpretation and Discussion

NOTE: Attach all data tables (1 to 5) and both the semi-log and linear graphs plotted in this
worksheet.

1. What does the pA2 value indicate? (1 mark)

…………………………………………………………………………………………
…………………………………………………………………………………………

2. Based on Schild plot analysis, what is your pA2 value? Show how this value is
obtained. (1, 1 mark)

pA2 = 9.15
Plot log(dose ratio-1) against negative log molar concentration of the antagonist(B) used(or directly against B) •

In summary, Schild plot analysis indicated that atropine is a potent competitive

acetylcholine antagonist characterized by a pA2 value of .
 3. Marks allocated for hand-drawn semi-log graph. (8 marks)
a. Appropriate title (1 m)
b. Clear legend (1 m)
Acetylcholine Maximum contractile force (gms)
Acetylcholine Maximum contractile force (gms)
Stock Vol. to add to Final 1 2 3 4 5 6 Average % Emax
Stock Vol. to add to Final % Emax
Normalized
conc. 10 mL organ conc. 6 Average Normalized
conc.
[M] 10bath
mL (uL)
organ conc. 1 2 3 4 5 (I)
[nM] (I)
[M] bath (uL) [nM]
1.00E-08 100 uL 1.00E-10 -0.059 -0.049 0.059 -0.020 0.195 -0.020 0.018 0.122
1.00E-08 100 uL 1.00E-10 0.410 0.391 0.449 0.420 0.381 0.469 0.420 2.989
1.00E-07 100 uL 1.00E-09 0.371 0.098 0.186 0.332 0.283 0.303 0.262 1.814
1.00E-07 100 uL 1.00E-09 2.999 2.980 3.175 3.107 3.205 3.283 3.125 22.241
1.00E-06 100 uL 1.00E-08 2.501 2.609 2.404 2.628 2.560 2.472 2.529 17.503
1.00E-06 100 uL 1.00E-08 10.787 10.532 10.513 10.611 10.699 10.542 10.614 75.544
1.00E-05 100 uL 1.00E-07 9.673 9.331 9.428 9.595 9.526 9.468 9.504 65.773
1.00E-05 100 uL 1.00E-07 13.933 13.991 13.874 13.952 13.923 13.942 13.936 99.187
1.00E-04 100 uL 1.00E-06 14.499 14.421 14.372 14.411 14.411 14.362 14.413 99.749
1.00E-04 100 uL 1.00E-06 14.050 14.069 14.060 14.021 14.060 14.040 14.050 100.000
1.00E-03 100 uL 1.00E-05 14.382 14.470 14.353 14.626 14.411 14.450 14.449 100.000

c. x- and y- axis correctly labeled (2 m)

d. EC50 values clearly indicated (4 m)

4. Marks allocated for hand-drawn linear-graph. (5 marks)

a. Appropriate title (1 m)
b. Clear legend (1 m)
c. x- and y- axis correctly labeled (2 m)
d. Schild slope drawn with X-intercept and slope value clearly indicated (1 m)

5. Marks allocated for Tables 1 - 5. (5 x 1 mark)

a. Consistency in data collection and correct calculations.

Total:

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 TABLE 3:
Results for Acetylcholine in the presence of 100nM Atropine

Acetylcholine Maximum contractile force (gms)

Stock Vol. to add to Final % Emax

conc. 10 mL organ conc. 1 2 3 4 5 6 Average Normalized
[M] bath (uL) [nM] (I)

1.00E-08 100 uL 1.00E-10 -0.195 0.391 0.186 0.009 -0.117 0.489 0.127 0.894

1.00E-07 100 uL 1.00E-09 -0.078 -0.059 0.127 0.009 0.098 0.029 0.021 0.148

1.00E-06 100 uL 1.00E-08 0.439 0.244 0.342 0.225 0.127 0.303 0.280 1.968

1.00E-05 100 uL 1.00E-07 2.706 2.774 2.892 2.843 2.745 2.735 2.783 19.558

1.00E-04 100 uL 1.00E-06 9.516 9.457 9.516 9.565 9.496 9.496 9.508 66.828

1.00E-03 100 uL 1.00E-05 12.594 12.574 12.721 12.594 12.653 12.672 12.635 88.808

1.00E-02 100 uL 1.00E-04 14.030 14.118 14.148 14.255 14.362 14.157 14.178 99.658

1.00E-01 100 uL 1.00E-03 14.235 14.030 14.372 14.196 14.167 14.362 14.227 100.000
 TABLE 4:
Results for Acetylcholine in the presence of 1000nM Atropine

Acetylcholine Maximum contractile force (gms)

Stock Vol. to add to Final % Emax

conc. 10 mL organ conc. 1 2 3 4 5 6 Average Normalized
[M] bath (uL) [nM] (I)

1.00E-08 100 uL 1.00E-10 0.049 0.668 0.029 0.059 0.098 0.088 0.165 1.184

1.00E-07 100 uL 1.00E-09 0.254 0.137 0.322 0.127 0.178 0.205 0.204 1.462

1.00E-06 100 uL 1.00E-08 0.166 0.342 0.234 0.186 0.127 0.147 0.200 1.437

1.00E-05 100 uL 1.00E-07 0.322 0.430 0.234 0.166 0.186 0.401 0.290 2.078

1.00E-04 100 uL 1.00E-06 2.423 2.667 2.472 2.350 2.335 2.394 2.440 17.498

1.00E-03 100 uL 1.00E-05 8.598 8.657 8.578 8.627 8.705 8.569 8.622 61.830

1.00E-02 100 uL 1.00E-04 11.207 11.363 11.255 11.265 11.197 11.275 11.260 80.746

1.00E-01 100 uL 1.00E-03 13.415 13.425 13.483 13.503 13.532 13.571 13.488 96.722

1.00E+00 100 uL 1.00E-02 13.845 13.903 13.962 14.060 14.089 13.813 13.945 100.000
 Table 5:
Calculation

Conc. ratio (cr)

Atropine concentration [B] EC50 value of Acetylcholine (cr-1) log (cr-1)
[A']/[A]
in the presence of
in Molar (M) in Log [B] Control, [A] (M)
atropine, [A'] (M)

0 - 3.162 x 10-9 3.162 x 10-9 1.000 0 -

1.0 x 10-8 -8 3.162 x 10-9 47.863 x 10-9 15.137 14.137 1.150

1.0 x 10-7 -7 3.162 x 10-9 416.869 x 10-9 131.837 130.837 2.117

1.0 x 10-6 -6 3.162 x 10-9 4073.802 x 10-9 1288.362 1287.362 3.110

Hand – Drawn concentration response curve

Log [B] Log (cr-1)

-8 1.15
-7 2.117
-6 3.11