title: Identification of Novel GPR55 Modulators Using Cell-Impedance-Based Label-Free Technology.
abstract: The orphan G protein-coupled receptor GPR55 has been proposed as a novel receptor of the endocannabinoid system. However, the validity of this categorization is still under debate mainly because of the lack of potent and selective agonists and antagonists of GPR55. Binding assays are not yet available for GPR55 screening, and discrepancies in GPR55 mediated signaling pathways have been reported. In this context, we have designed and synthesized novel GPR55 ligands based on a chromenopyrazole scaffold. Appraisal of GPR55 activity was accomplished using a label-free cell-impedance-based assay in hGPR55-HEK293 cells. The real-time impedance responses provided an integrative assessment of the cellular consequence to GPR55 stimulation taking into account the different possible signaling pathways. Potent GPR55 partial agonists (14b, 18b, 19b, 20b, and 21-24) have been identified; one of them (14b) being selective versus classical cannabinoid receptors. Upon antagonist treatment, chromenopyrazoles 21-24 inhibited lysophosphatidylinositol (LPI) effect. One of these GPR55 antagonists (21) is fully selective versus classic cannabinoid receptors. Compared to LPI, the predicted physicochemical parameters of the new compounds suggest a clear pharmacokinetic improvement.
authors.affiliation: Instituto de Química Médica, CSIC , Calle Juan de la Cierva, 3, 28006 Madrid, Spain.
authors.forename: Paula
authors.initials: P
authors.lastname: Morales
authors.affiliation: Department of Pharmacology and Toxicology, Medical Sciences Building, University of Toronto , 1 King's College Circle, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Lauren S
authors.initials: LS
authors.lastname: Whyte
authors.affiliation: Instituto de Química Médica, CSIC , Calle Juan de la Cierva, 3, 28006 Madrid, Spain.
authors.forename: Roberto
authors.initials: R
authors.lastname: Chicharro
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Complutense de Madrid , 28040 Madrid, Spain.|Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED) , 28040 Madrid, Spain.|Instituto Ramón y Cajal de Investigaciones Sanitarias (IRYCIS) , 28034 Madrid, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Gómez-Cañas
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Complutense de Madrid , 28040 Madrid, Spain.|Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED) , 28040 Madrid, Spain.|Instituto Ramón y Cajal de Investigaciones Sanitarias (IRYCIS) , 28034 Madrid, Spain.
authors.forename: M Ruth
authors.initials: MR
authors.lastname: Pazos
authors.affiliation: Instituto de Química Médica, CSIC , Calle Juan de la Cierva, 3, 28006 Madrid, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Goya
authors.affiliation: School of Biomolecular and Biomedical Science, University College Dublin , Dublin D4, Ireland.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Irving
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Complutense de Madrid , 28040 Madrid, Spain.|Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED) , 28040 Madrid, Spain.|Instituto Ramón y Cajal de Investigaciones Sanitarias (IRYCIS) , 28034 Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Fernández-Ruiz
authors.affiliation: Department of Pharmacology and Toxicology, Medical Sciences Building, University of Toronto , 1 King's College Circle, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Ruth A
authors.initials: RA
authors.lastname: Ross
authors.affiliation: Instituto de Química Médica, CSIC , Calle Juan de la Cierva, 3, 28006 Madrid, Spain.
authors.forename: Nadine
authors.initials: N
authors.lastname: Jagerovic
mesh_terms: D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D057075:Enzyme Assays / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D015394:Molecular Structure; D011720:Pyrazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D043882:Receptors, Cannabinoid; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship
chemical_list: C524885:GPR55 protein, human; D011720:Pyrazoles; D043882:Receptors, Cannabinoid; D043562:Receptors, G-Protein-Coupled