BU48: A Novel Buprenorphine Analog That Exhibits -Opioid- Mediated Convulsions but Not -Opioid-Mediated Antinociception in Mice 1 DANIEL C. BROOM, LI GUO, ANDREW COOP, 2 STEPHEN M. HUSBANDS, JOHN W. LEWIS, JAMES H. WOODS, and JOHN R. TRAYNOR Departments of Pharmacology (D.C.B., J.H.W., J.R.T.) and Psychology (J.H.W.), University of Michigan Medical School, Ann Arbor, Michigan; Department of Chemistry, University of Bristol, Bristol, United Kingdom (A.C., S.M.H., J.W.L.); and Department of Chemistry, Loughborough University, Loughborough, United Kingdom (L.G., J.R.T.) Accepted for publication May 25, 2000 This paper is available online at http://www.jpet.org ABSTRACT N-Cyclopropylmethyl-[7,8,2' ,3' ]-cyclohexano-1' [ S]-hy- droxy-6,14-endo-ethenotetrahydronororipavine (BU48) is a novel, ring-constrained analog of buprenorphine. In vivo, BU48 (0.1–10 mg/kg s.c.) produced brief, nonlethal convulsions in mice followed by brief Straub tail and a short period of cata- lepsy characteristic of BW373U86 and other nonpeptidic -re- ceptor agonists. BU48-induced convulsions were sensitive to antagonism by naltrindole (10 mg/kg s.c.) and were also pre- vented by administration of the putative 1 antagonist 7-ben- zylidenenaltrexone and the putative 2 antagonist naltriben, with the latter being more potent. In the abdominal stretch assay in the mouse, only low-efficacy antinociceptive activity of BU48 (0.1–10 mg/kg) was seen. This was reversed by the -opioid antagonist norbinaltorphimine (32 mg/kg s.c.) but not by the -opioid antagonist naltrindole (10 mg/kg s.c.). BU48 (10 mg/kg s.c.) acted as a -antagonist in this assay. In mouse brain homogenates, BU48 had high (nanomolar) binding affinity for all three opioid receptors in the order = . In vitro, the compound acted as a potent (EC 50 = 1.4 nM) -opioid agonist in the guinea pig ileum and a potent (EC 50 = 0.2 nM) -opioid agonist in the mouse vas deferens but showed partial agonist activity at the rat cloned -opioid (40%) and human cloned -opioid (59%) receptors with very low efficacy at the rat cloned -opioid receptor (10%); findings consistent with its in vivo profile. BU48 is the first described compound that produces -opioid-mediated convulsions without any evidence of -opi- oid-mediated antinociception and will be a useful tool in inves- tigations of the -opioid receptor. In the search for narcotic analgesic agents without the unwanted complications of -opioid receptor agonists such as morphine, research has focused on the -opioid receptor as a target for antinociceptive agents. This has culminated in the suggestion of -opioid receptor subtypes (for review, see Traynor and Elliot, 1993) and the discovery of the nonpeptide -opioid agonist BW373U86 and related compounds (e.g., Chang et al., 1993). BW373U86 produces antinociception without any apparent respiratory complications (Negus et al., 1994), although studies in mice showed a consistent and reproducible dose-dependent convulsive effect associated with BW373U86 administration (Comer et al., 1993). The convulsive activity of this compound has also been noted in squirrel monkeys (Dykstra et al., 1993; Pakarinen et al., 1995) and rhesus monkeys (Hong et al., 1998). The synthesis of SNC80, the methyl ether of the (+)-isomer of BW373U86 (Calderon et al., 1994), produced a compound more potent than BW373U86 as an antinociceptive agent. Received for publication January 14, 2000. 1 This work was supported by U.S. Public Health Service Grants DA00254, DA07315, and GM07767. Portions of this work were presented in abstract form at the 1999 meeting of the College on Problems of Drug Dependence, Acapulco, Mexico, June 12–17 (Broom et al., 2000). 2 Present address: Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 N. Pine St., Baltimore, MD 21201. ABBREVIATIONS: BW373U86, ()-[1(S*),2,5]-4-[[2,5-dimethyl-4-(2-propenyl)-1-piperazinyl](3-hydroxyphenyl)methyl]-N,N-diethylbenzamide hydrochloride; BNTX, 7-benzylidenenaltrexone; BU48, N-cyclopropylmethyl-[7,8,2',3']cyclohexano-1'[S]-hydroxy-6,14-endo-ethenotetrahy- dronororipavine; CI977, 5R-(5,7,8)-N-methyl-N-[7-(1-pyrrolidinyl-1-oxaspiro[4,5]dec-8-yl]-4-benzofuranacetamide; C6, C6 glioma cells transfected with the rat -receptor; CHO, Chinese hamster ovary; EEG, electroencephalographic; CNS, central nervous system; CHO-hkor, Chinese hamster ovary cells transfected with the human -receptor; C6, C6 glioma cells transfected with the rat -receptor; DAMGO, [D-Ala 2 ,N-Me-Phe 4 ,Gly 5 -ol]-enkephalin; DPDPE, [D-Pen 2 ,D-Pen 5 ]-enkephalin; GPI, myenteric plexus-longitudinal muscle of the guinea pig ileum; GTPS, guanosine-5'-O-(3-thio)triphosphate; MVD, mouse vas deferens; norBNI, norbinaltorphimine; NTB, naltriben; NTI, naltrindole; SNC80, (+)-4-[(R)--((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide; (-)-TAN-67, 2-methyl-4a-(3-hydroxyphen- yl)-1,2,3,4,4a,5,12,12a-octahydroquinolino[2,3,3-g]isoquinoline; U69593, 5,7,8-(+)-N-[7-(1-pyrrolidinyl)-1-oxaspiro[4,5]dec-8-yl]benzeneac- etamide; DMSO, dimethyl sulfoxide. 0022-3565/00/2943-1195$03.00/0 THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS Vol. 294, No. 3 Copyright © 2000 by The American Society for Pharmacology and Experimental Therapeutics 2493/846872 JPET 294:1195–1200, 2000 Printed in U.S.A. 1195 at ASPET Journals on October 26, 2016 jpet.aspetjournals.org Downloaded from