Novel Synthesis and Pharmacological Characterization of NOP Receptor Agonist 8-[(1S,3aS)-2,3,3a,4,5,6-Hexahydro-1H-phenalen-1-yl]-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (Ro 64-6198).

The nociceptin/orphanin FQ opioid peptide (NOP) receptor is a widely expressed GPCR involved in the modulation of pain, anxiety, and motor behaviors. Dissecting the functional properties of this receptor is limited by the lack of systemically active ligands that are brain permeant. The small molecule NOP receptor-selective, full agonist 8-[(1S,3aS)-2,3,3a,4,5,6-hexahydro-1H-phenalen-1-yl]-1-phenyl-1,3,8-triazaspiro[4.5]decan-4-one (Ro 64-6198) hydrochloride is an active, brain penetrant ligand, but its difficult and cost-prohibitive synthesis limits its widespread use and availability for animal studies. Here, we detail a more efficient and convenient method of synthesis, and use both in vitro and in vivo pharmacological assays to fully characterize this ligand. Specifically, we characterize the pharmacodynamics of Ro 64-6198 in cAMP and G-protein coupling in vitro and examine, for the first time, the effects of nociceptin/orphanin FQ and Ro 64-6198 in arrestin recruitment assays. Further, we examine the effects of Ro 64-6198 on analgesia, anxiety, and locomotor responses in vivo. This new synthesis and pharmacological characterization provide additional insights into the useful, systemically active, NOP receptor agonist Ro 64-6198.

[1]  M. Bruchas,et al.  Quantitative Signaling and Structure-Activity Analyses Demonstrate Functional Selectivity at the Nociceptin/Orphanin FQ Opioid Receptor , 2015, Molecular Pharmacology.

[2]  W. Spooren,et al.  Further characterization of the prototypical nociceptin/orphanin FQ peptide receptor agonist Ro 64-6198 in rodent models of conflict anxiety and despair , 2012, Psychopharmacology.

[3]  N. Murphy.  The nociceptin/orphanin FQ system as a target for treating alcoholism. , 2010, CNS & neurological disorders drug targets.

[4]  Christopher A. Podlesnik,et al.  The effects of nociceptin/orphanin FQ receptor agonist Ro 64-6198 and diazepam on antinociception and remifentanil self-administration in rhesus monkeys , 2010, Psychopharmacology.

[5]  J. Woods,et al.  Behavioral Effects of a Synthetic Agonist Selective for Nociceptin/Orphanin FQ Peptide Receptors in Monkeys , 2009, Neuropsychopharmacology.

[6]  E. Kostenis,et al.  Pharmacological profile of NOP receptors coupled with calcium signaling via the chimeric protein Gαqi5 , 2009, Naunyn-Schmiedeberg's Archives of Pharmacology.

[7]  David G. Lambert,et al.  The nociceptin/orphanin FQ receptor: a target with broad therapeutic potential , 2008, Nature Reviews Drug Discovery.

[8]  B. Kieffer,et al.  Effects of nociceptin/orphanin FQ receptor (NOP) agonist, Ro64-6198, on reactivity to acute pain in mice: comparison to morphine. , 2008, European journal of pharmacology.

[9]  L. Luttrell,et al.  Beta-arrestins 1 and 2 differentially regulate LPS-induced signaling and pro-inflammatory gene expression. , 2007, Molecular immunology.

[10]  G. Bakalkin,et al.  The Nociceptin/Orphanin FQ Receptor Agonist Ro 64-6198 Reduces Alcohol Self-Administration and Prevents Relapse-Like Alcohol Drinking , 2007, Neuropsychopharmacology.

[11]  D. Jessop,et al.  The nociceptin receptor antagonist [Nphe1,Arg14,Lys15]nociceptin/orphanin FQ-NH2 blocks the stimulatory effects of nociceptin/orphanin FQ on the HPA axis in rats , 2006, Neuroscience.

[12]  N. Maidment,et al.  The effect of a systemically active ORL-1 agonist, Ro 64-6198, on the acquisition, expression, extinction, and reinstatement of morphine conditioned place preference , 2005, Neuropharmacology.

[13]  Walter A. Korfmacher,et al.  Characterization of the nociceptin receptor (ORL-1) agonist, Ro64-6198, in tests of anxiety across multiple species , 2005, Psychopharmacology.

[14]  J. Felger,et al.  Nociceptin/Orphanin FQ Increases Anxiety-Related Behavior and Circulating Levels of Corticosterone During Neophobic Tests of Anxiety , 2004, Neuropsychopharmacology.

[15]  L. Terenius,et al.  Acquisition, Expression, and Reinstatement of Ethanol-Induced Conditioned Place Preference in Mice: Effects of Opioid Receptor-Like 1 Receptor Agonists and Naloxone , 2003, Journal of Pharmacology and Experimental Therapeutics.

[16]  J. Costentin,et al.  Nociceptin/orphanin FQ and related peptides reduce the increase in plasma corticosterone elicited in mice by an intracerebroventricular injection. , 2002, Life sciences.

[17]  J. Moreau,et al.  The orphanin receptor agonist RO 64-6198 does not induce place conditioning in rats , 2002, Neuroreport.

[18]  D. Lambert,et al.  Characterisation of the non-peptide nociceptin receptor agonist, Ro64-6198 in Chinese hamster ovary cells expressing recombinant human nociceptin receptors. , 2002, Life sciences.

[19]  G. Pasternak,et al.  The molecular and behavioral pharmacology of the orphanin FQ/nociceptin peptide and receptor family. , 2001, Pharmacological reviews.

[20]  H. Akil,et al.  Nociceptin/orphanin FQ regulates neuroendocrine function of the limbic–hypothalamic–pituitary–adrenal axis , 2001, Neuroscience.

[21]  G. Adam,et al.  Synthesis of (1S,3aS)-8-(2,3,3a,4,5, 6-hexahydro-1H-phenalen-1-yl)-1-phenyl-1,3,8-triaza-spiro[4. 5]decan-4-one, a potent and selective orphanin FQ (OFQ) receptor agonist with anxiolytic-like properties. , 2000, European journal of medicinal chemistry.

[22]  R. Houghten,et al.  Orphanin FQ/Nociceptin receptor binding studies , 2000, Peptides.

[23]  G. Adam,et al.  A synthetic agonist at the orphanin FQ/nociceptin receptor ORL1: anxiolytic profile in the rat. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Rees,et al.  G16 as a universal G protein adapter: implications for agonist screening strategies. , 1996, Trends in pharmacological sciences.

[25]  D. Grandy,et al.  Orphanin FQ: A Neuropeptide That Activates an Opioidlike G Protein-Coupled Receptor , 1995, Science.

[26]  Marc Parmentier,et al.  Isolation and structure of the endogenous agonist of opioid receptor-like ORL1 receptor , 1995, Nature.

[27]  C. Kozak,et al.  Molecular cloning, tissue distribution and chromosomal localization of a novel member of the opioid receptor gene family , 1994, FEBS letters.

[28]  T. Miyata,et al.  cDNA cloning and regional distribution of a novel member of the opioid receptor family , 1994, FEBS letters.

[29]  G Vassart,et al.  ORL1, a novel member of the opioid receptor family , 1994, FEBS letters.

[30]  M. Mortrud,et al.  Molecular cloning and tissue distribution of a putative member of the rat opioid receptor gene family that is not a mu, delta or kappa opioid receptor type. , 1994, FEBS letters.