LJMU Research Online Colestock, C, Wallach, J, Mansi, M, Filemban, N, Morris, H, Elliott, SP, Westphal, F, Brandt, SD and Adejare, A Syntheses, analytical and pharmacological characterizations of the “legal high” 4-[1-(3-methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo) and analogues http://researchonline.ljmu.ac.uk/id/eprint/6391/ Article Citation (please note it is advisable to refer to the publisher’s version if you intend to cite from this work) Colestock, C, Wallach, J, Mansi, M, Filemban, N, Morris, H, Elliott, SP, Westphal, F, Brandt, SD and Adejare, A (2017) Syntheses, analytical and pharmacological characterizations of the “legal high” 4-[1-(3- methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo) and analogues. Drug LJMU has developed LJMU Research Online for users to access the research output of the University more effectively. Copyright © and Moral Rights for the papers on this site are retained by the individual authors and/or other copyright owners. 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For more information please contact [email protected] http://researchonline.ljmu.ac.uk/ http://researchonline.ljmu.ac.uk/ Drug Testing and Analysis Syntheses, analytical and pharmacological characterizations of the “legal high” 4-[1-(3- methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo) and analogues For Peer Review Journal: Drug Testing and Analysis Manuscript ID DTA-17-0089.R1 Wiley - Manuscript type: Research Article Date Submitted by the Author: 05-May-2017 Complete List of Authors: Colestock, Tristan; University of the Sciences in Philadelphia Wallach, Jason; University of the Sciences, Mansi, Matt; University of the Sciences in Philadelphia Filemban, Nadine; University of the Sciences in Philadelphia Morris, Hamilton; New School for Social Research, Department of Anthropology Elliott, Simon; Alere Forensics (Forensics Ltd) Westphal, Dr. Folker; State Bureau of Criminal Investigation Schlesing Holstein, Nacotics/Toxicology Brandt, Simon; School of Pharmacy & Biomolecular Sciences , Liverpool John Moores University Adejare, Adeboye; University of the Sciences in Philadelphia New psychoactive substances, Dissociative anesthetics, NMDA receptor, Keywords: PCP, Arylcyclohexylmorpholines New psychoactive substances (NPS) are commonly referred to as “research chemicals”, “designer drugs” or “legal highs.” One NPS class is represented by dissociative anesthetics, which include analogues of the arylcyclohexylamine phencyclidine (PCP), ketamine, and diphenidine. A recent addition to the NPS market was 4-[1-(3- methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo), a morpholine analogue of 3-MeO-PCP. Although suspected to have dissociative effects in users, information about its pharmacological profile is not available. From clinical and forensic perspectives, detailed analytical data are needed for Abstract: identification, especially when facing the presence of positional isomers, as these are frequently unavailable commercially. This study presents the analytical and pharmacological characterization of 3-MeO-PCMo along with five additional analogues including the 2- and 4-MeO- isomers, 3,4- methylenedioxy-PCMo (3,4-MD-PCMo), 3-Me-PCMo and PCMo. All six arylcyclohexylmorpholines were synthesized and characterized by chromatographic, mass spectrometric and spectroscopic techniques. The three positional isomers could be differentiated and the identity of 3-MeO- PCMo obtained from an internet vendor was verified. All six compounds were also evaluated for affinity at 46 central nervous system receptors http://mc.manuscriptcentral.com/dta Page 1 of 25 Drug Testing and Analysis 1 2 3 4 including the N-methyl-D-aspartate receptor (NMDAR), an important target 5 for dissociative anesthetics such as PCP and ketamine. In vitro binding studies using [3H]-MK-801 in rat forebrain preparations revealed moderate 6 affinity for NMDAR in the rank order of 3-Me > 3-MeO >PCMo > 3,4-MD > 7 2-MeO > 4-MeO-PCMo. 3-MeO-PCMo was found to have moderate affinity 8 for NMDAR comparable to that of ketamine, and had an approximate 12- 9 fold lower affinity than PCP. These results support the anecdotal reports of 10 dissociative effects from 3-MeO-PCMo in humans. 11 12 13 14 15 16 17 18 For Peer Review 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 http://mc.manuscriptcentral.com/dta Drug Testing and Analysis Page 2 of 25 1 2 3 4 Syntheses, analytical and pharmacological characterizations of the “legal 5 high” 4-[1-(3-methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo) and 6 analogues 7 8 9 Tristan Colestock,a Jason Wallach, a Matt Mansi, a Nadine Filemban, a Hamilton 10 b c d e 11 Morris, Simon P. Elliott, Folker Westphal, Simon D. Brandt, Adeboye Adejare 12 a,* 13 14 15 a Department of Pharmaceutical Sciences, University of the Sciences, 600 South 43rd Street, 16 Philadelphia, PA 19104, USA 17 18 b For Peer Review 19 The New School for Social Research, Department of Anthropology, 66 West 12th Street, New York, NY 20 10011, USA 21 22 c Alere Forensics (Forensics Ltd), Malvern Hills Science Park, Geraldine Road, WR14 3SZ, UK 23 24 d State Bureau of Criminal Investigation Schleswig-Holstein, Section Narcotics/Toxicology, Mühlenweg 25 166, D-24116 Kiel, Germany 26 27 e 28 School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom Street, 29 Liverpool, L3 3AF, UK 30 31 32 *Corresponding author: Adeboye Adejare, Department of Pharmaceutical Sciences, 33 University of the Sciences, 600 S. 43 rd Street, Philadelphia PA 19128. E-mail: 34 35 [email protected] 36 37 38 39 40 41 Running title : Synthesis, analytical and pharmacological characterization of PCMo analogues 42 43 44 Keywords : New psychoactive substances; dissociative anesthetics; NMDA receptor; PCP; 45 46 arylcyclohexylmorpholines 47 48 49 50 51 52 53 54 55 56 57 58 59 60 1 http://mc.manuscriptcentral.com/dta Page 3 of 25 Drug Testing and Analysis 1 2 3 4 Abstract 5 6 New psychoactive substances (NPS) are commonly referred to as “research chemicals”, 7 “designer drugs” or “legal highs.” One NPS class is represented by dissociative anesthetics, 8 which include analogues of the arylcyclohexylamine phencyclidine (PCP), ketamine, and 9 diphenidine. A recent addition to the NPS market was 4-[1-(3- 10 methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo), a morpholine analogue of 3-MeO-PCP. 11 12 Although suspected to have dissociative effects in users, information about its pharmacological 13 profile is not available. From clinical and forensic perspectives, detailed analytical data are 14 needed for identification, especially when facing the presence of positional isomers, as these 15 are frequently unavailable commercially. This study presents the analytical and pharmacological 16 17 characterization of 3-MeO-PCMo along with five additional analogues including the 2- and 4- 18 MeO- isomers, 3,4-methylenedioxy-PCMoFor Peer (3,4-MD-PCMo) Review, 3-Me-PCMo and PCMo. All six 19 arylcyclohexylmorpholines were synthesized and characterized by chromatographic, mass 20 spectrometric and spectroscopic techniques. The three positional isomers could be 21 22 differentiated and the identity of 3-MeO-PCMo obtained from an internet vendor was verified. All 23 six compounds were also evaluated for affinity at 46 central nervous system receptors including 24 the N-methyl-D-aspartate receptor (NMDAR), an important target for dissociative anesthetics 25 such as PCP and ketamine. In vitro binding studies using [3H]-MK-801 in rat forebrain 26 27 preparations revealed moderate affinity for NMDAR in the rank order of 3-Me > 3-MeO >PCMo 28 > 3,4-MD > 2-MeO > 4-MeO-PCMo. 3-MeO-PCMo was found to have moderate affinity for 29 NMDAR comparable to that of ketamine, and had an approximate 12-fold lower affinity than 30 PCP. These results support the anecdotal reports of dissociative effects from 3-MeO-PCMo in 31 32 humans. 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 2 http://mc.manuscriptcentral.com/dta Drug Testing and Analysis Page 4 of 25 1 2 3 4 Introduction 5 [1] 6 A high number of new psychoactive substances (NPS) continue to be available from online 7 vendors and are sold as “research chemicals”. These chemicals are largely designed to by-pass 8 governmental restrictions on existing psychoactive drugs. Dissociative agents that target the N- 9 methyl-D-aspartate receptor (NMDAR) represent one of many available classes of compounds 10 that are encompassed by the NPS term. Substances with dissociative profile (Figure 1A) 11 12 comprise structural analogues of the arylcyclohexylamines such as phencyclidine (PCP), 13 ketamine and methoxetamine.[2] More recently, 1,2-diarylethylamines such as diphenidine and 14 its 2-methoxy analogue 2-MXP have also appeared.[3,4] 15 16 Substances that target the NMDAR are of interest for the development of treatment options for 17 conditions such as depression, neuropathic pain, and a variety of neurodegenerative disorders 18 [5-9] For Peer Review 19 and dementias. At the same time, a number of these substances