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Biotransformation of the New Synthetic Cannabinoid with an Alkene, MDMB-4en-PINACA, by Human Hepatocytes, Human Liver Microsomes, and Human Urine and Blood

The AAPS Journal volume 22, Article number: 13 (2020) Cite this article

Abstract

Although at a slower rate, new psychoactive substances continue to appear on the illicit drug market, challenging their detection in biological specimens by forensic and clinical toxicologists. Here, we report in vitro and in vivo metabolism of a new synthetic cannabinoid, methyl 3,3-dimethyl-2-[1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido] butanoate (MDMB-4en-PINACA). This is the first report on metabolism of a synthetic cannabinoid with an alkene functional group at the alkyl side chain. MDMB-4en-PINACA was incubated with both human hepatocytes and human liver microsomes (HLM) for up to 5 h and 1 h, respectively. The samples were analyzed by liquid chromatography-quadrupole time-of-flight mass spectrometry. An authentic human urine and a corresponding blood sample were analyzed to confirm the in vitro metabolites. A total of 32 metabolites were detected, of which 11 metabolites were detected in hepatocyte samples, 31 in HLM, 2 in urine, and 1 in blood. Analysis of the metabolites revealed that the main metabolic pathway of the terminal alkene group of the pentenyl side chain is dihydrodiol formation, most likely via epoxidation. The majority of the metabolites were generated from ester hydrolysis and/or dihydrodiol formation with further hydroxylation and/or dehydrogenation. Two most abundant metabolites in hepatocyte incubation samples, M8 (ester hydrolysis and dihydrodiol) and M30 (ester hydrolysis), coincided the two detected urinary metabolites. Based on the results, M8 and M30 are proposed to be appropriate urinary markers for MDMB-4en-PINACA intake for screening, while the inclusion of the parent drug itself and M29 (hydroxylation) may be useful for confirmation purposes.

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Acknowledgments

This work was supported by the Vinnova (the EUROSTARS Psychomics project, grant number 10628) and the Strategic Research Area in Forensic Sciences (Strategiområdet forensiska vetenskaper, grant number 2016:7) at Linköping University.

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Authors and Affiliations

  1. Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Artillerigatan 12, 587 58, Linköping, Sweden

    Shimpei Watanabe, Svante Vikingsson, Henrik Gréen & Robert Kronstrand

  2. Division of Drug Research, Department of Medical and Health Sciences, Linköping University, Linköping, Sweden

    Anna Åstrand, Henrik Gréen & Robert Kronstrand

Authors
  1. Shimpei Watanabe
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  2. Svante Vikingsson
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  3. Anna Åstrand
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  4. Henrik Gréen
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  5. Robert Kronstrand
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Correspondence to Shimpei Watanabe.

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Watanabe, S., Vikingsson, S., Åstrand, A. et al. Biotransformation of the New Synthetic Cannabinoid with an Alkene, MDMB-4en-PINACA, by Human Hepatocytes, Human Liver Microsomes, and Human Urine and Blood. AAPS J 22, 13 (2020). https://doi.org/10.1208/s12248-019-0381-3

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  • DOI: https://doi.org/10.1208/s12248-019-0381-3

KEY WORDS

  • Authentic human urine and blood
  • Human hepatocytes
  • Human liver microsomes
  • Metabolism
  • MDMB-4en-PINACA