title: In vitro functional characterization of novel nociceptin/orphanin FQ receptor agonists in recombinant and native preparations.
abstract: Nociceptin/Orphanin FQ (N/OFQ) regulates several biological functions via selective activation of the N/OFQ receptor (NOP). In this study novel nonpeptide NOP ligands were characterized in vitro in receptor binding and [35S]GTPγS stimulated binding in membranes of cells expressing human NOP and classical opioid receptors, calcium mobilization assay in cells coexpressing the receptors and chimeric G proteins, bioluminescence resonance energy transfer (BRET) based assay for studying NOP receptor interaction with G protein and arrestin, the electrically stimulated mouse vas deferens and the mouse colon bioassays. The action of the AT compounds were compared with standard NOP agonists (N/OFQ and Ro 65-6570) and the NOP selective antagonist SB-612111. AT compounds displayed high NOP affinity and behaved as NOP agonists in all the functional assays consistently showing the following rank order of potency AT-127≥AT-090≥AT-035>AT-004= AT-001. AT compounds behaved as NOP full agonists in the calcium mobilization and mouse colon assays and as partial agonists in the [35S]GTPγS and BRET assays. Interestingly AT-090 and AT-127, contrary to standard nonpeptide agonists that display G protein biased agonism, behaved as an unbiased agonists. AT-090 and AT-127 displayed higher NOP selectivity than Ro 65-6570 at native mouse receptors. AT-090 and AT-127 might be useful pharmacological tools for investigating the therapeutic potential of NOP partial agonists.
authors: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors: Federica
authors: F
authors: Ferrari
authors: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors: Maria Camilla
authors: MC
authors: Cerlesi
authors: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors: Davide
authors: D
authors: Malfacini
authors: Behavioral Pharmacology Laboratory, Department of Biophysics and Pharmacology, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
authors: Laila
authors: L
authors: Asth
authors: Behavioral Pharmacology Laboratory, Department of Biophysics and Pharmacology, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
authors: Elaine C
authors: EC
authors: Gavioli
authors: Astraea Therapeutics, LLC. 320 Logue Avenue, Mountain View, CA, USA.
authors: Blair V
authors: BV
authors: Journigan
authors: Astraea Therapeutics, LLC. 320 Logue Avenue, Mountain View, CA, USA.
authors: Uma Gayathri
authors: UG
authors: Kamakolanu
authors: Astraea Therapeutics, LLC. 320 Logue Avenue, Mountain View, CA, USA.
authors: Michael E
authors: ME
authors: Meyer
authors: Astraea Therapeutics, LLC. 320 Logue Avenue, Mountain View, CA, USA.
authors: Dennis
authors: D
authors: Yasuda
authors: SRI International, Biosciences Division, 333 Ravenswood Avenue, Menlo Park, CA, USA.
authors: Willma E
authors: WE
authors: Polgar
authors: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors: Anna
authors: A
authors: Rizzi
authors: Department of Chemical and Pharmaceutical Sciences and LTTA, University of Ferrara, Italy.
authors: Remo
authors: R
authors: Guerrini
authors: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors: Chiara
authors: C
authors: Ruzza
authors: Astraea Therapeutics, LLC. 320 Logue Avenue, Mountain View, CA, USA.
authors: Nurulain T
authors: NT
authors: Zaveri
authors: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy. Electronic address: g.calo@unife.it.
authors: Girolamo
authors: G
authors: Calo
mesh_terms: D000818:Animals; D016466:CHO Cells; D003106:Colon / Q000187:drug effects / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D003508:Cycloheptanes / Q000378:metabolism / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D008297:Male; D051379:Mice; D010880:Piperidines / Q000378:metabolism / Q000494:pharmacology*; D011957:Receptors, Opioid / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014649:Vas Deferens / Q000187:drug effects / Q000378:metabolism; D000094942:Nociceptin Receptor
chemical_list: D003508:Cycloheptanes; D008024:Ligands; D010880:Piperidines; D011957:Receptors, Opioid; D011994:Recombinant Proteins; C482404:cis-1-methyl-7-((4-(2,6-dichlorophenyl)piperidin-1-yl)methyl)-6,7,8,9-tetrahydro-5H-benzocyclohepten-5-ol; D000094942:Nociceptin Receptor
keywords: Bioluminescence resonance energy transfer; Calcium mobilization and mouse colon vas deferens and assays; NOP and classical opioid receptors; Receptor and [(35)S]GTPγS binding