Gemma Navarro1, Katia Varani2, Alejandro Lillo3, Fabrizio Vincenzi2, Rafael Rivas-Santisteban4, Iu Raïch4, Irene Reyes-Resina4, Carlos Ferreiro-Vera5, Pier Andrea Borea6, Verónica Sánchez de Medina5, Xavier Nadal7, Rafael Franco8. 1. Department of Biochemistry and Physiology. School of Pharmacy and Food Sciences, Universitat de Barcelona, Spain; Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CiberNed), Spain. 2. Department of Morphology, Surgery and Experimental Medicine, Ferrara University, Ferrara, Italy. 3. Department of Biochemistry and Physiology. School of Pharmacy and Food Sciences, Universitat de Barcelona, Spain; Department of Biochemistry and Molecular Biomedicine. Universitat de Barcelona, Spain. 4. Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CiberNed), Spain; Department of Biochemistry and Molecular Biomedicine. Universitat de Barcelona, Spain. 5. Phytoplant Research S.L., Córdoba, Spain. 6. Ferrara University, Ferrara, Italy. 7. Independent Researcher, Córdoba, Spain. 8. Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CiberNed), Spain; Department of Biochemistry and Molecular Biomedicine. Universitat de Barcelona, Spain. Electronic address: rfranco@ub.edu.
Abstract
BACKGROUND: Recent approved medicines whose active principles are Δ9Tetrahidrocannabinol (Δ9-THC) and/or cannabidiol (CBD) open novel perspectives for other phytocannabinoids also present in Cannabis sativa L. varieties. Furthermore, solid data on the potential benefits of acidic and varinic phytocannabinoids in a variety of diseases are already available. Mode of action of cannabigerol (CBG), cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabidivarin (CBDV) and cannabigerivarin (CBGV) is, to the very least, partial. HYPOTHESIS/ PURPOSE: Cannabinoid CB1 or CB2 receptors, which belong to the G-protein-coupled receptor (GPCR) family, are important mediators of the action of those cannabinoids. Pure CBG, CBDA, CBGA, CBDV and CBGV from Cannabis sativa L. are differentially acting on CB1 or CB2 cannabinoid receptors. STUDY DESIGN: Determination of the affinity of phytocannabinoids for cannabinoid receptors and functional assessment of effects promoted by these compounds when interacting with cannabinoid receptors. METHODS: A heterologous system expressing the human versions of CB1 and/or CB2 receptors was used. Binding to membranes was measured using radioligands and binding to living cells using a homogenous time resolved fluorescence resonance energy transfer (HTRF) assay. Four different functional outputs were assayed: determination of cAMP levels and of extracellular-signal-related-kinase phosphorylation, label-free dynamic mass redistribution (DMR) and ß-arrestin recruitment. RESULTS: Affinity of cannabinoids depend on the ligand of reference and may be different in membranes and in living cells. All tested phytocannabinoids have agonist-like behavior but behaved as inverse-agonists in the presence of selective receptor agonists. CBGV displayed enhanced potency in many of the functional outputs. However, the most interesting result was a biased signaling that correlated with differential affinity, i.e. the overall results suggest that the binding mode of each ligand leads to specific receptor conformations underlying biased signaling outputs. CONCLUSION: Results here reported and the recent elucidation of the three-dimensional structure of CB1 and CB2 receptors help understanding the mechanism of action that might be protective and the molecular drug-receptor interactions underlying biased signaling.
BACKGROUND: Recent approved medicines whose active principles are Δ9Tetrahidrocannabinol (Δ9-THC) and/or cannabidiol (CBD) open novel perspectives for other phytocannabinoids also present in Cannabis sativa L. varieties. Furthermore, solid data on the potential benefits of acidic and varinic phytocannabinoids in a variety of diseases are already available. Mode of action of cannabigerol (CBG), cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabidivarin (CBDV) and cannabigerivarin (CBGV) is, to the very least, partial. HYPOTHESIS/ PURPOSE: Cannabinoid CB1 or CB2 receptors, which belong to the G-protein-coupled receptor (GPCR) family, are important mediators of the action of those cannabinoids. Pure CBG, CBDA, CBGA, CBDV and CBGV from Cannabis sativa L. are differentially acting on CB1 or CB2 cannabinoid receptors. STUDY DESIGN: Determination of the affinity of phytocannabinoids for cannabinoid receptors and functional assessment of effects promoted by these compounds when interacting with cannabinoid receptors. METHODS: A heterologous system expressing the human versions of CB1 and/or CB2 receptors was used. Binding to membranes was measured using radioligands and binding to living cells using a homogenous time resolved fluorescence resonance energy transfer (HTRF) assay. Four different functional outputs were assayed: determination of cAMP levels and of extracellular-signal-related-kinase phosphorylation, label-free dynamic mass redistribution (DMR) and ß-arrestin recruitment. RESULTS: Affinity of cannabinoids depend on the ligand of reference and may be different in membranes and in living cells. All tested phytocannabinoids have agonist-like behavior but behaved as inverse-agonists in the presence of selective receptor agonists. CBGV displayed enhanced potency in many of the functional outputs. However, the most interesting result was a biased signaling that correlated with differential affinity, i.e. the overall results suggest that the binding mode of each ligand leads to specific receptor conformations underlying biased signaling outputs. CONCLUSION: Results here reported and the recent elucidation of the three-dimensional structure of CB1 and CB2 receptors help understanding the mechanism of action that might be protective and the molecular drug-receptor interactions underlying biased signaling.
Authors: Lyndsey L Anderson; Marika Heblinski; Nathan L Absalom; Nicole A Hawkins; Michael T Bowen; Melissa J Benson; Fan Zhang; Dilara Bahceci; Peter T Doohan; Mary Chebib; Iain S McGregor; Jennifer A Kearney; Jonathon C Arnold Journal: Br J Pharmacol Date: 2021-09-30 Impact factor: 9.473
Authors: Ayat Zagzoog; Kawthar A Mohamed; Hye Ji J Kim; Eunhyun D Kim; Connor S Frank; Tallan Black; Pramodkumar D Jadhav; Larry A Holbrook; Robert B Laprairie Journal: Sci Rep Date: 2020-11-23 Impact factor: 4.379
Authors: Rafael Franco; Gemma Navarro; Rafael Rivas-Santisteban; Alejandro Lillo; Jaume Lillo; Joan-Biel Rebassa; Joan S Contestí; Carlos A Saura Journal: Alzheimers Res Ther Date: 2021-11-08 Impact factor: 6.982