effects on CB1 Inverse Agonists Erin Cochran Endocannabinoid System Pharmacology Terms Basal or Constitutive activity physiological response in the absence of an agonist or inverse ID: 640920
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Cannabinoid-Receptor Interacting Protein 1a (CRIP1a) effects on CB1 Inverse Agonists
Erin CochranSlide2Slide3
Endocannabinoid SystemSlide4
Pharmacology Terms
Basal
or
Constitutive
activity- physiological response
in the absence of an agonist or inverse agonistAgonist-
ligand
that increases basal activity
Inverse
Agonist- a ligand that inhibits or eliminates basal activitySlide5
CB1 CRIP1a
Ahmed et al. 2015Slide6
Current Research
Smith et al. 2015
Full agonist: WIN
Partial agonists: MAE and THC
Inverse agonist: RIMSlide7
What Triggers CRIP1a Modulation?
Extreme ends of stimulation are modulated (full agonists and inverse agonists)
Moderate stimulation is not modulated (partial agonists)
Is CRIP1a modulation triggered by the intrinsic efficacy of a ligand or if
stimulation caused by the ligand
exceeds a range?Slide8
G-protein Coupled Receptor Activation
Harrison and
Traynor
2003Slide9
[35S]GTP
g
S
Binding Assay
Harrison and
Traynor
2003Slide10
Background Experiments
Transfect
Human Embryonic Kidney (HEK)
cell lines
hCB1-HEK
hCB1-HEK-CRIP1aReceptor Density[
3H]SR141716A binding assayControl
Basal binding
[
35
S]
GTP
gS binding assay without ligands presentBaselineSlide11
Inverse Agonists
Pertwee
2005Slide12
ExperimentCell Line Reactions
hCB1-HEK + inverse agonists +
[
35
S]GTP
gShCB1-HEK-CRIP1a + inverse agonists + [
35S]GTPg
S
Reaction stopped by vacuum filtration
Measure CB1 receptors bound with
[
35
S]GTPgS with scintillation counterSlide13
Data AnalysisNet Stimulation = Ligand Stimulation – Basal Binding
Percent Stimulation = (Net Stimulation / Basal Binding) X 100
Compare percent stimulation between hCB1-HEK and hCB1-HEK-CRIP1a
Compare percent stimulation between the ligands in hCB1-HEK-CRIP1a Slide14
Triggering CRIP1a Modulation
Intrinsic efficacy of ligand
All three inverse agonists stimulation will be reduced to the same degree
Reduction of Stimulation
SR141716A = AM251 = AM281
Extreme ends of range
Stimulation caused by the less potent inverse agonists will be reduced less than the more potent inverse agonists
Reduction of Stimulation
SR141716A > AM251 > AM281Slide15
Questions?