title: Pharmacological studies on the NOP and opioid receptor agonist PWT2-[Dmt1]N/OFQ(1-13).
abstract: An innovative chemical strategy named peptide welding technology (PWT) has been developed for the facile synthesis of tetrabranched peptides. [Dmt1]N/OFQ(1-13)-NH2acts as a universal agonist for nociceptin/orphanin FQ (N/OFQ) and classical opioid receptors. The present study investigated the pharmacological profile of the PWT derivative of [Dmt1]N/OFQ(1-13)NH2(PWT2-[Dmt1]) in several assays in vitro and in vivo after spinal administration in monkeys subjected to the tail withdrawal assay. PWT2-[Dmt1] mimicked the effects of [Dmt1]N/OFQ(1-13)-NH2displaying full agonist activity, similar affinity/potency and selectivity at human recombinant N/OFQ (NOP) and opioid receptors in receptor binding, stimulation of [35S]GTPγS binding, calcium mobilization in cells expressing chimeric G proteins, and BRET studies for measuring receptor/G-protein and receptor/β-arrestin 2 interaction. In vivo in monkeys PWT2-[Dmt1] elicited dose-dependent and robust antinociceptive effects being more potent and longer lasting than [Dmt1]N/OFQ(1-13)-NH2. The analgesic action of PWT2-[Dmt1] was sensitive to the NOP receptor antagonist J-113397, but not naltrexone. Thus, the present study demonstrated that the tetrabranched derivative of [Dmt1]N/OFQ(1-13)-NH2obtained with the PWT technology maintains the in vitro pharmacological profile of the parent peptide but displays higher potency and longer lasting action in vivo.
authors.affiliation: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors.forename: Maria Camilla
authors.initials: MC
authors.lastname: Cerlesi
authors.affiliation: Department of Physiology & Pharmacology, Wake Forest University School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Huiping
authors.initials: H
authors.lastname: Ding
authors.affiliation: Department of Cardiovascular Sciences, University of Leicester, Division of Anaesthesia, Critical Care and Pain Management, Leicester Royal Infirmary, Leicester LE2 7LX. UK.
authors.forename: Mark F
authors.initials: MF
authors.lastname: Bird
authors.affiliation: Department of Physiology & Pharmacology, Wake Forest University School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Norikazu
authors.initials: N
authors.lastname: Kiguchi
authors.affiliation: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Ferrari
authors.affiliation: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Malfacini
authors.affiliation: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Rizzi
authors.affiliation: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Ruzza
authors.affiliation: Department of Cardiovascular Sciences, University of Leicester, Division of Anaesthesia, Critical Care and Pain Management, Leicester Royal Infirmary, Leicester LE2 7LX. UK.
authors.forename: David G
authors.initials: DG
authors.lastname: Lambert
authors.affiliation: Department of Physiology & Pharmacology, Wake Forest University School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Mei-Chuan
authors.initials: MC
authors.lastname: Ko
authors.affiliation: Section of Pharmacology, Department of Medical Sciences, and National Institute of Neurosciences, University of Ferrara, Italy.
authors.forename: Girolamo
authors.initials: G
authors.lastname: Calo
authors.affiliation: Department of Chemical and Pharmaceutical Sciences and LTTA, University of Ferrara, Italy.
authors.forename: Remo
authors.initials: R
authors.lastname: Guerrini
mesh_terms: D000818:Animals; D016466:CHO Cells; D060326:Chemistry Techniques, Synthetic; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006801:Humans; D008253:Macaca mulatta; D008297:Male; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D011957:Receptors, Opioid / Q000819:agonists*; D011994:Recombinant Proteins / Q000378:metabolism; D000094942:Nociceptin Receptor
chemical_list: C584786:(Dmt(1))N-OFQ(1-13)-NH(2); D009842:Oligopeptides; D011957:Receptors, Opioid; D011994:Recombinant Proteins; D000094942:Nociceptin Receptor
keywords: Calcium mobilization; Monkey spinal analgesia; NOP and opioid receptors; PWT2-[Dmt(1)]N/OFQ(1–13); Receptor binding; Stimulation of [(35)S]GTPγS binding