Synthetic cannabinoids (SCs), mimicking the psychoactive effects of cannabis, consist of a vast array of structurally diverse compounds. A novel compound belonging to the SC family, (1-(cyclohexylmethyl)-1H-indol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone (named TMCP-CHM in this article) contains a cyclopropane ring that isomerizes during the smoking process, resulting in a ring-opened thermal degradant with a terminal double bond in its structure. Metabolites of TMCP-CHM were tentatively identified in vitro (after incubation of the parent substance with S9 pooled human liver fraction) and in vivo (rat experimental model) studies by accurate-mass liquid chromatography–tandem mass spectrometry (LC–MS/MS). For the identification of the degradant metabolites, and to study biotransformation of parent substance in the human, urine and hair samples from patients, who had ingested the compound and were subsequently admitted to hospital with drug intoxications, were analyzed. Products of mono-, di-, trihydroxylation, carboxylation, and carboxylation combined with hydroxylation of TMCP-CHM and its degradant were detected in human urine. Metabolism of the degradant included addition of water to the terminal double bond followed by dehydration and formation of a cyclic metabolite. Degradant metabolites prevailed in comparison with metabolites of the parent substance in each metabolite group examined, except carboxylation. N-Dealkylated metabolites found in human urine originated only from the degradant. Most of the hydroxy metabolites were detected in human urine in both the free form and as glucuronides. The detection of monohydroxylated (M1.1-M1.3, M/A1.10) and carboxylated/hydroxylated (M4.2, M/A4.3) metabolites of TMCP-CHM and the hydrated form of the monohydroxylated metabolite of the degradant was found to be convenient for routine analysis.

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Figure S1.

1H NMR spectrum of TMCP-CHM

Figure S2. ¹H NMR spectrum of TMCP-CHM (expansions)

Figure S3. ¹³C NMR spectrum of TMCP-CHM

Figure S4. GC-MS spectra of TMCP-CHM (a) and TMCP-CHM degradant (b)

Figure S5. Proposed scheme of fragmentation pathways of the TMCP-CHM and its degradant (GC-MS, EI)

Figure S6. LC-QTOF extracted ion chromatograms of the monohydroxylated metabolites of TMCP-CHM and its thermal degradant (MS², mass shift < 5 mDa). Ion transitions are shown on the right.

Table S1, LC-QTOF features of TMCP-CHM in vitro mono- and dihydroxylated metabolites

Figure S7. Dehydration of metabolites monohydroxylated on TMCP moiety. LC-QTOF extracted ion chromatograms (in vitro model, MS¹, mass shift < 5 mDa) (a), (b). Mass spectrum of V1.5 (c). Presumable structures of monohydroxylated metabolites and artificial dehydrated forms (d). Metabolites detected only in vitro are denoted as ‘V’

Figure S8. LC-QTOF spectra of monohydroxylated metabolites originated from TMCP-CHM degradant (M/A1.9), hydrated form (M/A1.10) and dihydrofuran form (M/A1.12)

Figure S9. LC-QTOF (MS², mass shift < 5 mDa) extracted ion chromatograms of the dihydroxylated metabolites of TMCP-CHM and its thermal degradant. Ion transitions are shown on the right. Metabolites detected only in vitro are denoted as ‘V’

Figure S10. Dehydration of dihydroxylated metabolites. LC-QTOF (in vitro model, MS¹, mass shift < 5 mDa) extracted ion chromatograms (a), (b). Mass spectra of V2.8 (c) and V2.9 (d). Presumable structures of dihydroxylated metabolites and artificial dehydrated forms (e). Metabolites detected only in vitro are denoted as ‘V’.

Figure S11. LC-QTOF (MS², mass shift < 5 mDa) extracted ion chromatograms of TMCP-CHM metabolites (carboxylated group). Ion transitions are shown on the right.

Figure S12. LC-QTOF (MS¹, mass shift < 5 mDa) extracted ion chromatograms of TMCP-CHM, its thermal degradant and their dealkylated metabolites. Values of ions m/z are shown on the right.

Figure S13. LC-QTOF (MS², mass shift < 5 mDa) extracted ion chromatograms (EICs) of TMCP-CHM metabolites (dealkylated with hydroxylation group). Ion transitions are shown on the right.

Figure S14. LC-QTOF (MS², mass shift < 5 mDa) extracted ion chromatograms (EICs) of TMCP-CHM metabolites (dealkylated with dihydroxylation group). Ion transitions are shown on the right.

Figure S15. LC-QTOF spectra of dealkylated metabolites

Table S2. LC-QTOF features of common N-dealkylated metabolites originating from TMCP-CHM (rat model) and XLR-11 (human urine)

Figure S16. LC-QTOF (MS², mass shift < 5 mDa) extracted ion chromatograms (EICs) of TMCP-CHM metabolites (trihydroxylation, M8 group) (a). Ion transitions are shown on the right. LC-QTOF spectra of trihydroxylated metabolites (b)

Table S3. LC-QTOF features of trihydroxylated metabolites of TMCP-CHM

Figure S17. Metabolites of M8 group (trihydroxylation) in the in vitro model. Dehydrated forms of trihydroxylated metabolites LC-QTOF (in vitro model, MS², mass shift < 5 mDa) extracted ion chromatograms (a). Ion transitions are shown on the right. LC-QTOF spectra of V8.1 (b) and V8.3 (c).

Figure S18. Metabolites of M/A9 group (hydration with oxidation to carboxyl and additional hydroxylation) in human urine. LC-QTOF (in vitro model, MS², mass shift < 5 mDa) extracted ion chromatograms (a). Ion transitions are shown on the right. LC-QTOF spectra of metabolites M/A9 group (b).

Table S4. LC-QTOF features of hydrated with oxidation to carboxyl and additional hydroxylation metabolites of TMCP-CHM degradant

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REFERENCES

1 PH Reggio (Ed). The cannabinoid receptors. New York, NY: Humana Press; 2009 10.1007/978-1-59745-503-9.
2Banister SD, Connor M. The chemistry and pharmacology of synthetic cannabinoid receptor agonists as new psychoactive substances: origins. Handb Exp Pharmacol. 2018; 252: 165-190. https://doi.org/10.1007/164_2018_143
3Banister SD, Connor M. The chemistry and pharmacology of synthetic cannabinoid receptor agonist new psychoactive substances: evolution. Handb Exp Pharmacol. 2018; 252: 191-226. https://doi.org/10.1007/164_2018_144
4 EMCDDA. European drug report: trends and developments. 2018. http://www.emcdda.europa.eu/system/files/publications/8585/20181816_TDAT18001ENN_PDF.pdf. Accessed Jan 30, 2019.
5Shevyrin V, Melkozerov V, Nevero A, Eltsov O, Morzherin Y, Shafran Y. Identification and analytical properties of new synthetic cannabimimetics bearing 2,2,3,3-tetramethylcyclopropanecarbonyl moiety. Forensic Sci Int. 2013; 226(1-3): 62-73. https://doi.org/10.1016/j.forsciint.2012.12.009
6Choi H, Heo S, Kim E, Hwang BY, Lee C, Lee J. Identification of (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone and its 5-pentyl fluorinated analog in herbal incense seized for drug trafficking. Forensic Toxicol. 2013; 31(1): 86-92. https://doi.org/10.1007/s11419-012-0170-5
7Frost JM, Dart MJ, Tietje KR, et al. Indol-3-ylcycloalkyl ketones: effects of N1 substituted indole side chain variations on CB(2) cannabinoid receptor activity. J Med Chem. 2010; 53(1): 295-315. https://doi.org/10.1021/jm901214q
8Banister SD, Stuart J, Kevin RC, et al. Effects of bioisosteric fluorine in synthetic cannabinoid designer drugs JWH-018, AM-2201, UR-144, XLR-11, PB-22, 5F-PB-22, APICA, and STS-135. ACS Chem Nerosci. 2015; 6(8): 1445-1458. https://doi.org/10.1021/acschemneuro.5b00107
9 WHO. Expert committee on drug dependence. UR-144 critical review report. 39th Meeting. Agenda Item 4.11. 2017. https://www.who.int/medicines/access/controlled-substances/CriticalReview_UR144.pdf. Accessed May 10, 2019.
10 WHO. Expert committee on drug dependence. XLR-11 critical review report. 39th Meeting. Agenda Item 4.12. 2016. https://www.who.int/medicines/access/controlled-substances/4.12_XLR-11_CritReview.pdf. Accessed May 10, 2019.
11Kavanagh P, Grigoryev A, Savchuk S, Mikhura I, Formanovsky A. UR-144 in products sold via the internet: identification of related compounds and characterization of pyrolysis products. Drug Test Anal. 2013; 5(8): 683-692. https://doi.org/10.1002/dta.1456
12Kaizaki-Mitsumoto A, Hataoka K, Funada M, Odanaka Y, Kumamoto H, Numazawa S. Pyrolysis of UR-144, a synthetic cannabinoid, augments an affinity to human CB1 receptor and cannabimimetic effects in mice. J Toxicol Sci. 2017; 42(3): 335-341. https://doi.org/10.2131/jts.42.335
13Grigoryev A, Kavanagh P, Melnik A, Savchuk S, Simonov A. Gas and liquid chromatography-mass spectrometry detection of the urinary metabolites of UR-144 and its major pyrolysis product. J Anal Toxicol. 2013; 37(5): 265-276. https://doi.org/10.1093/jat/bkt028
14Karinen R, Tuv SS, Øiestad EL, Vindenes V. Concentrations of APINACA, 5F-APINACA, UR-144 and its degradant product in blood samples from six impaired drivers compared to previous reported concentrations of other synthetic cannabinoids. Forensic Sci Int. 2015; 246: 98-103. https://doi.org/10.1016/j.forsciint.2014.11.012
15Adamowicz P, Lechowicz W. The influence of synthetic cannabinoid UR-144 on human psychomotor performance – a case report demonstrating road traffic risks. Traffic Inj Prev. 2015; 16(8): 754-759. https://doi.org/10.1080/15389588.2015.1018990
16Amaratunga P, Thomas C, Lemberg BL, Lemberg D. Quantitative measurement of XLR11 and UR-144 in oral fluid by LC-MS-MS. J Anal Toxicol. 2014; 38(6): 315-321. https://doi.org/10.1093/jat/bku040
17Adamowicz P, Zuba D, Sekuła K. Analysis of UR-144 and its pyrolysis product in blood and their metabolites in urine. Forensic Sci Int. 2013; 233(1-3): 320-327. https://doi.org/10.1016/j.forsciint.2013.10.005
18Kanamori T, Kanda K, Yamamuro T, et al. Detection of main metabolites of XLR-11 and its thermal degradation product in human hepatoma HepaRG cells and human urine. Drug Test Anal. 2015; 7(4): 341-345. https://doi.org/10.1002/dta.1765
19Sobolevsky T, Prasolov I, Rodchenkov G. Detection of urinary metabolites of AM-2201 and UR-144, two novel synthetic cannabinoids. Drug Test Anal. 2012; 4(10): 745-753. https://doi.org/10.1002/dta.1418
20Hutter M, Broecker S, Kneisel S, Franz F, Brandt SD, Auwarter V. Metabolism of nine synthetic cannabinoid receptor agonists encountered in clinical casework: major in vivo phase I metabolites of AM-694, AM-2201, JWH-007, JWH-019, JWH-203, JWH-307, MAM-2201, UR-144 and XLR-11 in human urine using LC-MS/MS. Curr Pharm Biotechnol. 2018; 19(2): 144-162. https://doi.org/10.2174/1389201019666180509163114
21Watanabe S, Kuzhiumparambil U. Fu S. Structural elucidation of metabolites of synthetic cannabinoid UR-144 by Cunninghamella elegans using nuclear magnetic resonance (NMR) spectroscopy. AAPS j. 2018; 20(2): 42. https://doi.org/10.1208/s12248-018-0209-6.
22Watanabe S, Kuzhiumparambil U, Nguyen MA, Cameron J, Fu S. Metabolic profile of synthetic cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans. AAPS j. 2017; 19(4): 1148-1162. https://doi.org/10.1208/s12248-017-0078-4
23Park M, Yeon S, Lee J, In S. Determination of XLR-11 and its metabolites in hair by liquid chromatography-tandem mass spectrometry. J Pharm Biomed Anal. 2015; 114: 184-189. https://doi.org/10.1016/j.jpba.2015.05.022
24Wohlfarth A, Pang S, Zhu M, et al. First metabolic profile of XLR-11, a novel synthetic cannabinoid, obtained by using human hepatocytes and high-resolution mass spectrometry. Clin Chem. 2013; 59(11): 1638-1648. https://doi.org/10.1373/clinchem.2013.209965
25Jang M, Kim IS, Park YN, et al. Determination of urinary metabolites of XLR-11 by liquid chromatography–quadrupole time-of-flight mass spectrometry. Anal Bioanal Chem. 2016; 408(2): 503-516. https://doi.org/10.1007/s00216-015-9116-1
26Nielsen LM, Holm NB, Olsen L, Linnet K. Cytochrome P450-mediated metabolism of the synthetic cannabinoids UR-144 and XLR-11. Drug Test Anal. 2016; 8(8): 792-800. https://doi.org/10.1002/dta.1860
27Nikitin EV, Grigoryev AM, Labutin AV, et al. Detection of metabolites of TMCP-CHMINACA, the new psychoactive substance, in rat urine and serum by liquid chromatography-mass spectrometry. Mass-Spektrometria. 2018; 15(3): 162-171 (in Russian). https://doi.org/10.25703/MS.2018.15.34

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Tentative identification of the metabolites of (1-(cyclohexylmethyl)-1H-indol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone, and the product of its thermal degradation, by in vitro and in vivo methods

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Tentative identification of the metabolites of (1-(cyclohexylmethyl)-1H-indol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone, and the product of its thermal degradation, by in vitro and in vivo methods

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