In vitro functional characterization of novel nociceptin/orphanin FQ receptor agonists in recombinant and native preparations

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• 作者：Federica Ferrari^a ; Maria Camilla Cerlesi^a ; Davide Malfacini^a ; Laila Asth^b ; Elaine C. Gavioli^b ; Blair V. Journigan^c ; Uma Gayathri Kamakolanu^c ; Michael E. Meyer^c ; Dennis Yasuda^c ; Willma E. Polgar^d ; Anna Rizzi^a ; Remo Guerrini^e ; Chiara Ruzza^a ; Nurulain T. Zaveri^c ; Girolamo Calo^a ; ^{g.calo@unife.it}
• 关键词：NOP and classical opioid receptors ; Bioluminescence resonance energy transfer ; Calcium mobilization and mouse colon vas deferens and assays ; Receptor and [35S]GTPγS binding
• 刊名：European Journal of Pharmacology
• 出版年：2016
• 出版时间：15 December 2016
• 年：2016
• 卷：793
• 期：Complete
• 页码：1-13
• 全文大小：1315 K
• 卷排序：793

文摘

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.