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In vitro metabolic profiles of adamantyl positional isomers of synthetic cannabinoids

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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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Acknowledgements

This study was funded by the Health and Labor Sciences Research Grants 2015–2017 and 2018–2020 to K. Kitaichi (Research on Regulatory Science of Pharmaceuticals and Medical Devices, No. 201523005A and 201824009A) and a grant-in-aid for Scientific Research to K. Kitaichi (No. 17K08417). A portion of this work was supported by the domestically programmed grant for the regional society from the Gifu Prefectural Research Institute for Health and Environmental Sciences. We also acknowledge the Gifu Regional Consortium on the Development of Analytical Procedures for Legal Highs operated by Gifu Pharmaceutical University and the Gifu Prefectural Research Institute for Health and Environmental Sciences for supervising the experimental protocols.

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Author notes

  1. Tetsuro Ito

    Present address: Laboratory of Pharmacognosy, Department of Pharmacy, Faculty of Pharmacy, Gifu University of Medical Science, 4-3-3 Nijigaoka, Kani, Gifu, 509-0293, Japan

Authors and Affiliations

  1. Laboratory of Pharmaceutics, Department of Biomedical Pharmaceutics, Gifu Pharmaceutical University, 1-25-4, Daigaku-nishi Gifu, Gifu, 501-1196, Japan

    Natsuki Kadomura, Tetsuro Ito, Hidenobu Kawashima, Takaya Matsuhisa, Tomoe Kinoshita, Midori Soda & Kiyoyuki Kitaichi

  2. Research Institute for Health and Environmental Sciences, Gifu Prefectural Government, 1-1, Naka Fudogaoka, Kakamigahara, 504-0838, Japan

    Natsuki Kadomura, Tetsuro Ito, Hidenobu Kawashima, Takaya Matsuhisa, Tomoe Kinoshita, Erina Kohyama, Takaharu Iwaki & Hiroyuki Nagai

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Kadomura, N., Ito, T., Kawashima, H. et al. In vitro metabolic profiles of adamantyl positional isomers of synthetic cannabinoids. Forensic Toxicol 39, 26–44 (2021). https://doi.org/10.1007/s11419-020-00538-7

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