Drug Testing and Analysis Return of the lyserga mides. Part I: Analytical and behavioral characterization of 1-propionyl-d-lysergic acid diethylamide (1P-LSD) For Peer Review Journal: Drug Testing and Analysis Manuscript ID: DTA-15-0251.R1 Wiley - Manuscript type: Research Article Date Submitted by the Author: n/a Complete List of Authors: Brandt, Simon; School of Pharmacy & Biomolecular Sciences , Liverpool John Moores University Kavanagh, Pierce; Trinity Centre for Health Sciences, Department of Pharmacology and Therapeutics Westphal, Folker; State Bureau of Criminal Investigation Schleswig- Holstein, Section Narcotics/Toxicology Stratford, Alexander; Synex Ltd, Elliott, Simon; ROAR Forensics, Hoang, Khoa; University of the Sciences, Department of Chemistry and Biochemistry Wallach, Jason; University of the Sciences, Department of Pharmaceutical Sciences Halberstadt, Adam; University of California San Diego, Department of Psychiatry New psychoactive substances, LSD, psychedelics, 5-HT2A receptor, head- Keywords: twitch response 1-Propionyl-d-lysergic acid diethylamide hemitartrate (1P-LSD) has become available as a ‘research chemical’ in form of blotters and powdered material. This non-controlled derivative of d-lysergic acid diethylamide (LSD) has previously not been described in the published literature despite being closely related to 1-acetyl-LSD (ALD-52), which was developed in the 1950s. This study describes the characterization of 1P-LSD in comparison with LSD using various chromatographic, mass spectrometric methods and nuclear magnetic resonance spectroscopy. An important feature common to LSD and other serotonergic hallucinogens is that they produce 5-HT2A- Abstract: receptor activation and induce the head-twitch response (HTR) in rats and mice. In order to assess whether 1P-LSD displays LSD-like properties and activates the 5-HT2A receptor, male C57BL/6J mice were injected with vehicle (saline) or 1P-LSD (0.025‒0.8 mg/kg, IP) and HTR assessed for 30 min using magnetometer coil recordings. It was found that 1P-LSD produced a dose-dependent increase in HTR counts, and that it had ~38% (ED50 = 349.6 nmol/kg) of the potency of LSD (ED50 = 132.8 nmol/kg). Furthermore, the HTR was abolished when 1P-LSD administration followed pre-treatment with the selective 5-HT2A receptor antagonist M100907 (0.1 mg/kg, SC), which confirms that the behavioral response is mediated by http://mc.manuscriptcentral.com/dta Page 1 of 30 Drug Testing and Analysis 1 2 3 4 activation of the 5-HT2A receptor. These results indicate that 1P-LSD 5 produces LSD-like effects in mice, consistent with its classification as a serotonergic hallucinogen. Nevertheless, the extent to which 1P-LSD might 6 show psychoactive effects in humans similar to LSD remains to be 7 investigated. 8 9 10 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 30 1 2 3 Return of the lysergamides. Part I: Analytical and behavioral 4 characterization of 1-propionyl-d-lysergic acid diethylamide (1P- 5 6 LSD) 7 8 Simon D. Brandt, a,* Pierce V. Kavanagh, b Folker Westphal, c Alexander 9 d e f g 10 Stratford, Simon P. Elliott, Khoa Hoang, Jason Wallach, Adam L. h 11 Halberstadt, 12 13 14 a School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom 15 Street, Liverpool L3 3AF, UK 16 17 b 18 Department ofFor Pharmacology Peer and Therapeutics, Review School of Medicine, Trinity Centre for 19 Health Sciences, St. James Hospital, Dublin 8, Ireland 20 c 21 State Bureau of Criminal Investigation Schleswig-Holstein, Section Narcotics/Toxicology, 22 Mühlenweg 166, D-24116 Kiel, Germany 23 24 d Synex Ltd, 67-68 Hatton Garden, N13 4BS, London, UK 25 26 e ROAR Forensics, Malvern Hills Science Park, Geraldine Road, WR14 3SZ, UK 27 28 f Department of Chemistry and Biochemistry, University of the Sciences, Philadelphia, PA 29 19104, USA 30 31 g 32 Department of Pharmaceutical Sciences, Philadelphia College of Pharmacy, University of 33 the Sciences, Philadelphia, PA 19104, USA 34 h 35 Department of Psychiatry, University of California San Diego, La Jolla, CA 92093-0804, 36 USA 37 38 39 * Correspondence to: Simon D. Brandt, School of Pharmacy and Biomolecular Sciences, 40 Liverpool John Moores University, Byrom Street, Liverpool, L3 3AF, UK. E-Mail: 41 [email protected] 42 43 44 45 Running title: Analytical and behavioral characterization of 1-propionyl-LSD 46 47 48 Keywords: New psychoactive substances; LSD; 5-HT 2A receptor; lysergamides; 49 psychedelics 50 51 52 53 54 55 56 57 58 59 60 1 http://mc.manuscriptcentral.com/dta Page 3 of 30 Drug Testing and Analysis 1 2 3 Abstract 4 5 1Propionyl-d-lysergic acid diethylamide hemitartrate (1P-LSD) has become available 6 7 as a ‘research chemical’ in form of blotters and powdered material. This non- 8 controlled derivative of d-lysergic acid diethylamide (LSD) has previously not been 9 described in the published literature despite being closely related to 1acetyl-LSD 10 (ALD-52), which was developed in the 1950s. This study describes the 11 characterization of 1P-LSD in comparison with LSD using various chromatographic, 12 13 mass spectrometric methods and nuclear magnetic resonance spectroscopy. An 14 important feature common to LSD and other serotonergic hallucinogens is that they 15 produce 5-HT 2A -receptor activation and induce the head-twitch response (HTR) in 16 rats and mice. In order to assess whether 1P-LSD displays LSD-like properties and 17 activates the 5-HT 2A receptor, male C57BL/6J mice were injected with vehicle (saline) 18 or 1P-LSD (0.025‒0.8For mg/kg, Peer IP) and HTR Reviewassessed for 30 min using magnetometer 19 20 coil recordings. It was found that 1P-LSD produced a dose-dependent increase in 21 HTR counts, and that it had ~38% (ED 50 = 349.6 nmol/kg) of the potency of LSD 22 (ED 50 = 132.8 nmol/kg). Furthermore, the HTR was abolished when 1P-LSD 23 administration followed pre-treatment with the selective 5-HT 2A receptor antagonist 24 M100907 (0.1 mg/kg, SC), which confirms that the behavioral response is mediated 25 by activation of the 5-HT receptor. These results indicate that 1P-LSD produces 26 2A 27 LSD-like effects in mice, consistent with its classification as a serotonergic 28 hallucinogen. Nevertheless, the extent to which 1P-LSD might show psychoactive 29 effects in humans similar to LSD remains to be investigated. 30 31 Introduction 32 33 [1] 34 It is perhaps fair to consider that the synthesis and discovery of the psychoactive 35 properties of d-lysergic acid diethylamide (LSD) [2] (Figure 1) in 1943 triggered an 36 avalanche of investigations that continue to capture the imagination of researchers 37 across all disciplines.[3-10] Although the pharmacology and properties of LSD have 38 been investigated in many studies, major questions still remain unanswered and are 39 [11-16] 40 expected to occupy the attention of researchers in the future. 41 42 Reports have been published indicating LSD may possess therapeutic efficacy in 43 patients suffering from disorders such as anxiety, alcoholism, cluster headaches, and 44 autism, but unfortunately most of this evidence is anecdotal in nature or confounded 45 [17-19] 46 by methodological shortcomings. Importantly, although most clinical work with 47 LSD ceased in the late 1960s, human trials have cautiously resumed during the last 48 few years.[20-23] 49 50 A range of lysergamide derivatives have been prepared to explore their molecular 51 [24-32] pharmacology (e.g. ) but the extent to which these show psychoactive properties 52 [7] 53 in humans is not always clear. In recent years, several lysergamide derivatives 54 have been distributed as new psychoactive substances or “research chemicals” in 55 the UK and Europe.[33] For example, lysergic acid 2,4-dimethylazetidide (LSZ)[31] and [24,30] 56 N6-allyl-6norlysergic acid diethylamide (AL-LAD) are two lysergamide 57 derivatives with LSD-like effects in animals that originated from academic research 58 59 60 2 http://mc.manuscriptcentral.com/dta Drug Testing and Analysis Page 4 of 30 1 2 3 and have been available for purchase in powdered and blotter form. Another closely 4 related derivative with modification at the indole nitrogen is 1acetyl-LSD (ALD-52) 5 (Figure 1). Synthesis of ALD-52 was first reported in 1957 (e.g. [34] ) and it was found 6 to be psychoactive in humans [35-38] but it is not clear whether ALD-52 was also sold in 7 8 the UK. Recent changes in UK legislation, however, precluded the open sale of [39] 9 several lysergamides, including ALD-52, LSZ and AL-LAD. In response to those 10 legal restrictions, 1-propionyl-LSD (Figure 1), also known as 1P-LSD, became 11 available as a “research chemical” either as powdered material or on blotters. 12 Although an assortment of LSD derivatives substituted at the 1-position have been 13 described (e.g. [34,37,40-42] ), it is noteworthy that chemical, analytical or pharmacological 14 15 data related to 1-propionyl-LSD appear to be absent from the literature. 16 17 Because of the difficulty associated with studying hallucinogens in humans, animal 18 behavioral modelsFor are often Peer used to investigate Review the pharmacology of hallucinogenic 19 drugs. One behavioral model that has been widely adopted is the head twitch 20 response (HTR), a paroxysmal side-to-side head movement induced by 5-HT 21 2A 22 agonists in rats and mice.