NEW CANNABINOIDTHERAPEUTICSTamema ChoudhuryVincent LiDavid McNee
The endocannabinoid system• Established cannabinoid receptors: CB1 and CB2.• Other orphan receptors (e.g. GPR55) as potential targets.• Endogenous ligands include Anandamide (AEA) and 2- arachidonoyl glycerol (2-AG).• Variety of potential clinical uses including pain management, obesity and schizophrenia.
Fatty acid amino hydrolase (FAAH) FAAH Inactive Endocannabinoids Metabolites Orthosteric binding site FAAH INHIBITORS CB1 Pain & inflammation Receptor Depression α β γ Signals which control neuronal No side effects? excitability
EndocannabinoidsAllosteric site • Anandamide • 2AG Orthosteric binding site Novel small molecules allosteric CB1 Receptor α γ β Signals which control neuronal Endogenous Agonist excitabilityEffect Time Allosteric inhibitorEffect Time
Method• Mouse Brain Membrane Preparation • Homogenised and centrifuged several times with different speeds at 4°C. • Protein concentration determined using spectrophotometry. Agonist• [35S]GTPγS Functional Assay GPCR • Measures the activity of G-protein through the accumulation of α γ β membrane-bound Gα[35S]GTPγS. GDP • Beta particles emitted are detected with a liquid scintillation counter. GTPγS• Equilibrium Binding Assay (Affinity) • Measures displacement of [3H]CP55940 at the CB1 receptors orthosteric site. • Beta particles emitted are detected with a liquid scintillation counter.
Summary• Cannabinoid compounds are an important area of pharmacological discovery. The compounds tested may be worthy of further investigation to produce novel clinical therapeutics. • JK263-2 as a treatment for chronic pain. • ORG27569 to assist combating obesity.• Special thanks to Gemma Baillie, Professor Ross and others in the Cannabinoid Group.