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01-01-2021 | Cannabinoid | Original Article

In vitro metabolic profiles of adamantyl positional isomers of synthetic cannabinoids

Authors: Natsuki Kadomura, Tetsuro Ito, Hidenobu Kawashima, Takaya Matsuhisa, Tomoe Kinoshita, Midori Soda, Erina Kohyama, Takaharu Iwaki, Hiroyuki Nagai, Kiyoyuki Kitaichi

Published in: Forensic Toxicology | Issue 1/2021

Abstract

Purpose

Illegal use of synthetic cannabinoids (SCs) is a serious problem worldwide. Legal regulation of SCs requires fundamental analytical studies regarding the differentiation of potential structural isomers. Accumulation of SC metabolic profiles is also essential for forensic investigation because SCs are immediately metabolized after intake. Thus, we investigated the in vitro metabolism of N-adamantyl-1-(tetrahydropyran-4-ylmethyl)-1H-indazole-3-carboxamide isomers (ATHs) using human liver microsomes (HLMs). Moreover, we validated the applicability of the isomeric differentiation by investigation of N-adamantyl-1-(4-fluorobenzyl)-1H-indazole-3-carboxamide isomers (AFUs).

Methods

Metabolites were collected at designated time points during the incubation period with HLMs for up to 180 min. The structures of the metabolites were annotated on the basis of mass spectroscopic evidence obtained by liquid chromatography–ion trap–time of flight mass spectrometry.

Results

The secondary stage mass (MS2) spectra obtained from the protonated molecules revealed a clear difference in both ATHs and their major metabolites because of the stability of the adamantyl (AD) cation. In HLMs, ATHs were quickly metabolized, and hydroxylation of the AD ring was deduced as the major metabolic pathway. The major metabolites of ATH 1 and ATH 2 after 180 min showed dihydroxylation and monohydroxylation of the AD ring. The AFUs showed analytical and metabolic profiles similar to those of the ATHs described above.

Conclusions

We characterized the metabolism of ATHs for the first time and discriminated between the two isomers by mass spectrometric analysis of either the parent compounds or their major metabolites. Our investigation of AFUs also demonstrated a useful method for distinguishing between AD isomers.

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Diao X, Huestis MA (2017) Approaches, challenges, and advances in metabolism of new synthetic cannabinoids and identification of optimal urinary marker metabolites. Clin Pharmacol Ther 101:239–253. https://doi.org/10.1002/cpt.534CrossRefPubMed

Underwood E (2015) A new drug war. Science 347:469–473CrossRef

Diao X, Huestis MA (2019) New synthetic cannabinoids metabolism and strategies to best identify optimal marker metabolites. Front Chem 7:1–15. https://doi.org/10.3389/fchem.2019.00109CrossRef

Gandhi AS, Zhu M, Pang S, Wohlfarth A, Scheidweiler KB, Liu HF, Huestis MA (2013) First characterization of AKB-48 metabolism, a novel synthetic cannabinoid, using human hepatocytes and high-resolution mass spectrometry. AAPS J 15:1091–1098. https://doi.org/10.1208/s12248-013-9516-0CrossRefPubMedPubMedCentral

Asada A, Doi T, Tagami T, Takeda A, Sawabe Y (2017) Isomeric discrimination of synthetic cannabinoids by GC-EI-MS: 1-adamantyl and 2-adamantyl isomers of N-adamantyl carboxamides. Drug Test Anal 9:378–388. https://doi.org/10.1002/dta.2124CrossRefPubMed

European Monitoring Centre for Drugs and Drug Addiction (2016) EMCDDA-europol, 2015 annual report on the implementation of Council Decision 2005/387/JHA, implementation reports, Publications Office of the European Union, Luxembourg. http://www.emcdda.europa.eu/system/files/publications/2880/TDAS16001ENN.pdf. Accessed 7 Sept 2016

List of “Designated Substances” in Japan. https://www.mhlw.go.jp/content/11120000/000563116.pdf. Accessed 24 Nov 2019

Murakami T, Iwamuro Y, Ishimaru R, Chinaka S, Takayama N, Hasegawa H (2018) Differentiation of AB-FUBINACA and its five positional isomers using liquid chromatography–electrospray ionization-linear ion trap mass spectrometry and triple quadrupole mass spectrometry. Forensic Toxicol 36:351–358. https://doi.org/10.1007/s11419-018-0410-4CrossRefPubMedPubMedCentral

Diao X, Scheidweiler KB, Wohlfarth A, Zhu M, Pang S, Huestis MA (2016) Strategies to distinguish new synthetic cannabinoid FUBIMINA (BIM-2201) intake from its isomer THJ-2201: metabolism of FUBIMINA in human hepatocytes. Forensic Toxicol 34:256–267. https://doi.org/10.1007/s11419-016-0312-2CrossRefPubMedPubMedCentral

10.

Vikingsson S, Josefsson M, Gréen H (2015) Identification of AKB-48 and 5F-AKB-48 metabolites in authentic human urine samples using human liver microsomes and time of flight mass spectrometry. J Anal Toxicol 39:426–435. https://doi.org/10.1093/jat/bkv045CrossRefPubMed

11.

Erratico C, Negreira N, Norouzizadeh H, Covaci A, Neels H, Maudens K, van Nuijs ALN (2015) In vitro and in vivo human metabolism of the synthetic cannabinoid AB-CHMINACA. Drug Test Anal 7:866–876. https://doi.org/10.1002/dta.1796CrossRefPubMed

12.

De Brabanter N, Esposito S, Tudela E, Lootens L, Meuleman P, Leroux-Roels G, Deventer K, Van Eenoo P (2013) In vivo and in vitro metabolism of the synthetic cannabinoid JWH-200. Rapid Commun Mass Spectrom 27:2115–2126. https://doi.org/10.1002/rcm.6673CrossRefPubMed

13.

Baranczewski P, Stańczak A, Sundberg K, Svensson R, Wallin Å, Jansson J, Garberg P, Postlind H (2006) Introduction to in vitro estimation of metabolic stability and drug interactions of new chemical entities in drug discovery and development. Pharmacol Rep 58:453–472PubMed

14.

Holm NB, Pedersen AJ, Dalsgaard PW, Linnet K (2015) Metabolites of 5F-AKB-48, a synthetic cannabinoid receptor agonist, identified in human urine and liver microsomal preparations using liquid chromatography high-resolution mass spectrometry. Drug Test Anal 7:199–206. https://doi.org/10.1002/dta.1663CrossRefPubMed

15.

Takayama T, Suzuki M, Todoroki K, Inoue K, Min JZ, Kikura-Hanajiri R, Goda Y, Toyo’oka T (2014) UPLC/ESI-MS/MS-based determination of metabolism of several new illicit drugs, ADB-FUBINACA, AB-FUBINACA, AB-PINACA, QUPIC, 5F-QUPIC and α-PVT, by human liver microsome. Biomed Chromatogr 28:831–838. https://doi.org/10.1002/bmc.3155CrossRefPubMed

16.

Savchuk S, Appolonova S, Pechnikov A, Rizvanova L, Shestakova K, Tagliaro F (2017) In vivo metabolism of the new synthetic cannabinoid APINAC in rats by GC–MS and LC–QTOF-MS. Forensic Toxicol 35:359–368. https://doi.org/10.1007/s11419-017-0364-yCrossRef

17.

Kusano M, Zaitsu K, Yamanaka M, Hisatsune K, Asano T, Taki K, Hayashi Y, Tsuchihashi H, Ishii A (2016) Development of a mass spectrometric hydroxyl-position determination method for the hydroxyindole metabolites of JWH-018 by GC-MS/MS. J Mass Spectrom 51:350–357. https://doi.org/10.1002/jms.3761CrossRefPubMed

18.

Belal TS, Thaxton-Weissenfluh A, DeRuiter J, Smith F, Abiedalla Y, Abdel-Hay KM, Clark CR (2018) Differentiation of methylated indole ring regioisomers of JWH-007: GC–MS and GC–IR studies. Forensic Chem 7:1–9. https://doi.org/10.1016/j.forc.2017.11.001CrossRef

19.

Castaneto MS, Wohlfarth A, Pang S, Zhu M, Scheidweiler KB, Kronstrand R, Huestis MA (2015) Identification of AB-FUBINACA metabolites in human hepatocytes and urine using high-resolution mass spectrometry. Forensic Toxicol 33:295–310. https://doi.org/10.1007/s11419-015-0275-8CrossRef

20.

Andersson M, Diao X, Wohlfarth A, Scheidweiler KB, Huestis MA (2016) Metabolic profiling of new synthetic cannabinoids AMB and 5F-AMB by human hepatocyte and liver microsome incubations and high-resolution mass spectrometry. Rapid Commun Mass Spectrom 30:1067–1078. https://doi.org/10.1002/rcm.7538CrossRefPubMed

21.

Mardal M, Dalsgaard PW, Qi B, Mollerup CB, Annaert P, Linnet K (2018) Metabolism of the synthetic cannabinoids AMB-CHMICA and 5C-AKB48 in pooled human hepatocytes and rat hepatocytes analyzed by UHPLC-(IMS)-HR-MSE. J Chromatogr B: Anal Technol Biomed Life Sci 1083:189–197. https://doi.org/10.1016/j.jchromb.2018.03.016CrossRef

22.

Diao X, Wohlfarth A, Pang S, Scheidweiler KB, Huestis MA (2016) High-resolution mass spectrometry for characterizing the metabolism of synthetic cannabinoid THJ-018 and its 5-fluoro analog THJ-2201 after incubation in human hepatocytes. Clin Chem 62:157–169. https://doi.org/10.1373/clinchem.2015.243535CrossRefPubMed

23.

Vikingsson S, Gréen H, Brinkhagen L, Mukhtar S, Josefsson M (2016) Identification of AB-FUBINACA metabolites in authentic urine samples suitable as urinary markers of drug intake using liquid chromatography quadrupole tandem time of flight mass spectrometry. Drug Test Anal 8:950–956. https://doi.org/10.1002/dta.1896CrossRefPubMed

24.

Hwang J, Hwang J, Ganganna B, Song I, Heo MY, Ahn SH, Lee J (2018) Metabolic and pharmacokinetic characterization of a new synthetic cannabinoid APINAC in rats. Forensic Toxicol 36:88–101. https://doi.org/10.1007/s11419-017-0387-4CrossRef

25.

Holm NB, Nielsen LM, Linnet K (2015) CYP3A4 mediates oxidative metabolism of the synthetic cannabinoid AKB-48. AAPS J 17:1237–1245. https://doi.org/10.1208/s12248-015-9788-7CrossRefPubMedPubMedCentral

26.

Rasul G, Olah GA, Prakash GKS (2004) Density functional theory study of adamantanediyl dications C₁₀H₁₄²⁺ and protio-adamantyl dications C₁₀H₁₆²⁺. Proc Natl Acad Sci 101:10868–10871. https://doi.org/10.1073/pnas.0404137101CrossRefPubMed

27.

Kevin RC, Lefever TW, Snyder RW, Patel PR, Fennell TR, Wiley JL, McGregor IS, Thomas BF (2017) In vitro and in vivo pharmacokinetics and metabolism of synthetic cannabinoids CUMYL-PICA and 5F-CUMYL-PICA. Forensic Toxicol 35:333–347. https://doi.org/10.1007/s11419-017-0361-1CrossRefPubMedPubMedCentral

28.

Chauret N, Gauthier A, Nicoll-Griffith DA (1998) Effect of common organic solvents on in vitro cytochrome P450-mediated metabolic activities in human liver microsomes. Drug Metab Dispos 26:1–4PubMed

29.

Drug Enforcement Administration, Department of Justice. (2019) Schedules of controlled substances: temporary placement of 5F-EDMB-PINACA, 5F-MDMB-PICA, FUB-AKB48, 5F-CUMYL-PINACA, and FUB-144 into Schedule I. Fed Regist 84:15505–15511. https://www.govinfo.gov/content/pkg/FR-2019-04-16/pdf/2019-07460.pdf. Accessed 16 Apr 2019

30.

Hess C, Schoeder CT, Pillaiyar T, Madea B, Müller CE (2016) Pharmacological evaluation of synthetic cannabinoids identified as constituents of spice. Forensic Toxicol 34:329–343. https://doi.org/10.1007/s11419-016-0320-2CrossRefPubMedPubMedCentral

31.

Diao X, Scheidweiler KB, Wohlfarth A, Pang S, Kronstrand R, Huestis MA (2016) In vitro and in vivo human metabolism of synthetic cannabinoids FDU-PB-22 and FUB-PB-22. AAPS J 18:455–464. https://doi.org/10.1208/s12248-016-9867-4CrossRefPubMedPubMedCentral

Title: In vitro metabolic profiles of adamantyl positional isomers of synthetic cannabinoids
Authors: Natsuki Kadomura
Tetsuro Ito
Hidenobu Kawashima
Takaya Matsuhisa
Tomoe Kinoshita
Midori Soda
Erina Kohyama
Takaharu Iwaki
Hiroyuki Nagai
Kiyoyuki Kitaichi
Publication date: 01-01-2021
Publisher: Springer Singapore
Keyword: Cannabinoid
Published in: Forensic Toxicology / Issue 1/2021
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI: https://doi.org/10.1007/s11419-020-00538-7

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

In vitro metabolic profiles of adamantyl positional isomers of synthetic cannabinoids

Abstract

Purpose

Methods

Results

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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