13_14 MD

Dr. Collonese
9/5/14
10:00-11:00 AM
Notetaker 35


Pharmacodynamics Workshop
Note: There were a few additional slides and questions added to the presentation that
were not given in the hard-out or notes posted on Blackboard.

Slide 21: Compound N acts at the MC 4 receptor as a full agonist. You more or less call
something a full agonist because it more or less as the same effect as the endogenous
ligand. Compound O acts at the MC4 receptor as a partial agonist because it has a lower
efficacy than the endogenous ligand (alphaMSH) Compared to compound N, compound
O has a lower efficacy. N and O have the same potency because you can calculate the
half-way point from where it asymptotes because both compounds bind fully to the
receptor, but its just not fully activating the receptor. In this case, we measure the
halfway points for both O and N and trace it down to the X axis. They both have the
same potency. You cannot measure affinity from this graph, but under the same
conditions you would assume O and N have the same binding affinities. You are unable
to determine from this graph if Compound P acts at the MC4 receptor as a competitive
or noncompetitive antagonist. Compound P has no effect on this cell, at any
concentration. Compound P does bind to the receptor, but it does so without any
change on the cell. A competitive antagonist binds to the receptor site and hinders the
endogenous ligand (e.g. alphaMSH) from binding. Compound Q dropped below the
baseline, so based on this, Q acts at the MC4 receptor as an inverse agonist. Inverse
agonists are annoying, and can happen when you have a baseline production of the
product you want, like in this case with 200 M cAMP. It turns out in the case of
inverse agonists that this baseline production was actually due to the MC4 receptor. So
compound Q bound somewhere on the GPCR and induced a conformational change
which hindered the MC4 receptor and reduced the amount of baseline cAMP produced.
This is a hard concept to grasp. You dont call it an antagonist, because you arent
blocking anything, it is just that when nothing is bound, it is making a baseline amount
of material- but when you bind the receptor (with compound Q), some of the
production is shut down.

Question; is alphaMSH present in these graphs?
alphaMSH is present in the black line, but under totally different conditions we have
tested N, O, P, and Q. With that being said, you are unable to determine whether any of
the compounds are a competitive or uncompetitive antagonist.



Slide 22: not covered

Slide 23: These graded-dose response curves are generated using inhibitors with
alphaMSH being present. This study shows drugs that bind the receptor but do not
have any effect on the receptor themselves. We want to see how they affect the
endogenous signal using the following graphs.

You measure with MSH alone, MSH with inhibitor R, and MSH with inhibitor S and you
get the response curves. Graded dose response curves use a concentration, or
something with units- not a percentage. Based on the data, Compound R is a
competitive antagonist and a full antagonist. You are measuring a response to different
concentrations of alpha MSH not of the inhibitor itself. If you were measuring the
inhibitor itself, this graph would actually show compound R being an agonist. When
looking at these graphs determine if they are measuring the effectiveness of the ligand,
or the effectiveness of the thing with the inhibitor. Its a competitive antagonist
because it can eventually be overcome and the curve shifted to the right. The inhibitor
has affected the potency of the main drug. Based on this Data, Compound S is a non-
competitive antagonist. This is because it decreased the efficacy of the primary drug.


Slide 24: Competitive antagonists are drugs that look like the original compound and
bind at the same spot but just dont have an effect on the cell. These antagonists
basically just reduce the bound concentration of the drug at the receptor. Non-
competitive antagonist are irreversible and can either bind at the same site, or they
can bind to another site and cause a conformational change at the receptor, in turn
hindering ligand binding. When non-competitive antagonists bind to the receptor, they
cannot be outcompeted by high concentrations of endogenous ligand, therefore they
cannot be removed from the receptor. They take the receptors out of commission so
you can no longer get the same size of an effect. Inhibitors do not change the affinity of
the ligand for the receptor. However, according to the graph the potency was reduced,
but the important thing to consider is that the efficacy was reduced (he seemed to be
confused by this concept).


Slide 25: not covered

Slide 26: not covered

Slide 27: The last thing we are going to cover is the third type of response curve is the
Exponential- Dose response curve, which measures how closely receptors bind to their
targets. You need to review that there are both physiologic and nonphysiologic
receptors. The question is asked: Which of the following could serve as a receptor for a
drug? This question includes physiologic and non-physiologic receptors. All the
options can serve as a receptor because they are proteins or endogenous peptides. The
examples were: COX1, K Channel in beta cells, Dopamine reuptake pump in neurons,
and Dopamine D2 in neurons. The dopamine D2 receptor is the only one that can
serve as a physiologic receptor. This is because the dopamine D2 receptor is the only
one that is a receptor for a ligand, while the others are nonphysiologic receptors
including voltage-gated channel pumps and non-ligand gated ion channels.


Slide 28: not covered

Slide 29: Basic binding equation, you can get a measurement of the tightness you have
bound between the ligand and the receptor. He then read the equation out loud. With
that you are able to measure the tightness that you have.

Slide 31: In this next case we want to take the MC4 receptor and measure how much
ligand is bound, so we add radioactive alphaMSH and you get a curve that looks like
this (see slide 32). We want to measure bMAX ( which is the maximum bound) for the
alphaMSH to this receptor.


Slide 32: Based on the graph what is the maximal binding ( bMAX) for alphaMSH
binding to the brain membrane MC4 receptors? The answer is 65. It is at the maximal
binding line so where the line asymptotes. The Kd (affinity) for alphaMSH binding to
brain membranes is estimated as 3 if you find the asymptote which we agreed was 65,
half of that would be in the thirties, cross to the right and drop down to the
concentration, so we end up at 3 nM for the Kd.


Slide 33/34: Now I am going to test your knowledge of affinity by giving you an
inverted view. This slide shows a Competitive Receptor binding study. We have the
alphaMSH at one concentration at the bMAX and give increasing concentrations of
known competitors (competitive antagonists). What is measured is the amount of
alphaMSH bound to the receptors in the presence of different concentrations of these
receptors. We have people graph it out on graph paper and then we can also show you
in color.


Slide 35: Z does not compete with alpha MSH, but Y and X both hinder alphaMSH from
binding to the receptor and they do so with different efficacies. According to the
binding graphs, competitor X has a higher affinity than competitor Y, as it binds more
tightly to the MC4 receptor. Competitor Z is not a competitive antagonist because it is
not affecting the binding of alphaMSH on the MC4 receptor. Competitor X has a higher
affinity is all we can say for certain, and we know this because there is an inferred
LOWER Kd. At a lower concentration it displaces more alphaMSH, therefore it has a
higher affinity.