New Cannabinoid Therapeutics

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New Cannabinoid Therapeutics

  1. 1. NEW CANNABINOIDTHERAPEUTICSTamema ChoudhuryVincent LiDavid McNee
  2. 2. 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.
  3. 3. Orthosteric site ORTHOSTERIC BLOCKERS Endocannabinoids Phytocannabinoids Synthetic small molecules • Anandamide • Δ9THC • Agonists • 2-AG • Inverse Agonists Obesity & metabolic syndrome Nicotine addiction Mental illness Drug abuse Orthosteric binding site Suicide & depression CB1 Receptor ORTHOSTERIC ACTIVATORS α γ β Signals which Pain & inflammation control neuronal Multiple sclerosis excitability Euphoria, Memory loss, Hallucination
  4. 4. 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
  5. 5. 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
  6. 6. 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.
  7. 7. Results – JK263-2 1µm JK263-2 JK-263-2 100 DMSO 120 CP55940 100 % Stimulation [35S]GTPγS Binding % displacement of [3H]CP55940 80 80 60 60 40 20 40 0 -20 20 -40 -60 0 -80 -20 -100 -10 -9 -8 -7 -6 -5 -4 -11 -10 -9 -8 -7 -6 -5 -4 CP55940 log concentration (M) log concentration (M)
  8. 8. Results – JK263-2 (cont.) 140 DMSO 100nM JK_263-2 % Stimulation [35S]GTPγS Binding 120 100 80 60 40 20 0 -20 -10 -9 -8 -7 -6 -5 -4 AEA log concentration (M)
  9. 9. Results ORG-27569 DMSO ORG27569 60 120 1µm ORG27569 CP55940 100 % Stimulation [35S]GTPγS Binding % displacement of [3H]CP55940 80 40 60 40 20 20 0 -20 0 -40 -60 -80 -20 -100 -10 -9 -8 -7 -6 -5 -4 -10 -9 -8 -7 -6 -5 -4 CP55940 log concentration (M) log concentration (M)
  10. 10. Key findings• JK263-2: Increases both affinity and efficacy. Potential allosteric enhancer.• ORG-27569: Increases affinity whilst inhibiting efficacy. Potential allosteric inhibitor.
  11. 11. 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.

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