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In vivo and in vitro metabolism of the novel synthetic cannabinoid 5F-APINAC

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Abstract

Purpose

New generations of synthetic cannabinoids (SCs) have recognized strong psychoactive effects, rapidly becoming potent drugs of abuse. Characterization of metabolites associated with SCs is useful for their subsequent detection in biological samples. We aimed to investigate the metabolism of 5F-APINAC, a novel SC, in human liver microsomes (HLMs) (in vitro) and in a rat model (in vivo).

Methods

For the in vitro study, the standard solution of 5F-APINAC was incubated with HLMs for 1 h at 37 °C. In the in vivo study, rats received 15 mg/kg of 5F-APINAC diluted in ethanol or in dimethyl sulfoxide, while a vehicle control group received placebo. Urine was collected 3, 6, and 24 h after administration. The metabolic characterization was performed using liquid chromatography–ion trap mass spectrometry and liquid chromatography–quadrupole time-of-flight mass spectrometry.

Results

In total, 15 metabolites associated with 5F-APINAC were tentatively identified. The metabolites were classified as: 1-adamantol metabolites, oxidative defluorination metabolites, and N-fluoropentylindazole-3-carboxylic acid metabolites. The modifications included ester hydrolysis, mono-, di-, and trihydroxylation of adamantyl ring and N-fluoropentylindazole moiety, oxidation (carbonyl formation) of the N-fluoropentyl side chain, oxidative loss of fluorine, and glucuronidation, as well as combinations thereof. The predominant metabolic reaction was ester hydrolysis in both in vitro and in vivo experiments and formation of M9 (5-fluoropentylindazole-3-carboxylic acid). However, the most recommendable metabolites for proving 5F-APINAC consumption in urine were M4.1, M7, and M13. No metabolites were detected in rat’s urine 24 h after drug administration.

Conclusions

The discovered metabolites are proposed to be incorporated into routine screening analytical methods as the urine markers of 5F-APINAC consumption. This is the first report to demonstrate the metabolism of 5F-APINAC to our knowledge.

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Acknowledgements

We appreciate the general input received during the preparation of this article from Professor Helgi Schiöth, Uppsala University, Sweden. This study was funded by Project 5-100 Sechenov University Grant.

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Authors and Affiliations

  1. Laboratory of Pharmacokinetics and Metabolomic Analysis, Institute of Translational Medicine and Biotechnology, I.M. Sechenov First Moscow State Medical University, 2-4 Bolshaya Pirogovskaya St., Moscow, 119991, Russia

    Svetlana A. Appolonova, Ksenia M. Shestakova, Natalia V. Mesonzhnik, Alex Brito, Roman M. Kuznetsov, Pavel A. Markin, Natalia L. Bochkareva, Giacomo Musile, Franco Tagliaro & Sergey A. Savchuk

  2. Unit of Forensic Medicine, Department of Diagnostics and Public Health, University of Verona, 37134, Verona, Italy

    Covadonga Palacio, Giacomo Musile & Franco Tagliaro

  3. PhD Program in Nanosciences and Advanced Technologies, University of Verona, 37134, Verona, Italy

    Covadonga Palacio, Ksenia M. Shestakova & Pavel A. Markin

  4. Bruker Ltd, Moscow, 119017, Russia

    Dmitry Burmykin & Maxim Ovcharov

  5. Russian Center of Forensic Medical Expertise, Moscow, 125284, Russia

    Sergey A. Savchuk

Authors
  1. Svetlana A. Appolonova
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  2. Covadonga Palacio
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  3. Ksenia M. Shestakova
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  4. Natalia V. Mesonzhnik
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  8. Natalia L. Bochkareva
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  11. Giacomo Musile
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  12. Franco Tagliaro
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  13. Sergey A. Savchuk
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Corresponding author

Correspondence to Svetlana A. Appolonova.

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Conflict of interest

DB and MO are affiliated with the company Bruker Ltd, Moscow, Russia. SAA, CP, KMS, NVM, AB, RMK, PAM, NLB, GM, FT, and SAS have no conflict of interest to declare.

Ethical approval

All animal procedures were approved by the Institutional Animal Care and Ethical Committee of the Research Center for Molecular Diagnostics and Therapy at Sechenov University, Moscow, in accordance with principles of good laboratory practice [OECD Principles on GLP. C (97)186 Final], following the European Union directive principles of laboratory animal care guidelines (2010/63/EU). This article does not contain any studies with human participants performed by any of the authors.

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Appolonova, S.A., Palacio, C., Shestakova, K.M. et al. In vivo and in vitro metabolism of the novel synthetic cannabinoid 5F-APINAC. Forensic Toxicol 38, 160–171 (2020). https://doi.org/10.1007/s11419-019-00503-z

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  • DOI: https://doi.org/10.1007/s11419-019-00503-z

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