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Metabolism of synthetic cannabinoids PB-22 and its 5-fluoro analog, 5F-PB-22, by human hepatocyte incubation and high-resolution mass spectrometry

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Abstract

Background

PB-22 (1-pentyl-8-quinolinyl ester-1H-indole-3-carboxylic acid) and 5F-PB-22 (1-(5-fluoropentyl)-8-quinolinyl ester-1H-indole-3-carboxylic acid) are new synthetic cannabinoids with a quinoline substructure and the first marketed substances with an ester bond linkage. No human metabolism data are currently available, making it difficult to document PB-22 and 5F-PB-22 intake from urine analysis, and complicating assessment of the drugs’ pharmacodynamic and toxicological properties.

Methods

We incubated 10 μmol/l PB-22 and 5F-PB-22 with pooled cryopreserved human hepatocytes up to 3 h and analyzed samples on a TripleTOF 5600+ high-resolution mass spectrometer. Data were acquired via TOF scan, followed by information-dependent acquisition triggered product ion scans with mass defect filtering (MDF). The accurate mass full scan MS and MS/MS metabolite datasets were analyzed with multiple data processing techniques, including MDF, neutral loss and product ion filtering.

Results

The predominant metabolic pathway for PB-22 and 5F-PB-22 was ester hydrolysis yielding a wide variety of (5-fluoro)pentylindole-3-carboxylic acid metabolites. Twenty metabolites for PB-22 and 22 metabolites for 5F-PB-22 were identified, with the majority generated by oxidation with or without glucuronidation. For 5F-PB-22, oxidative defluorination occurred forming PB-22 metabolites. Both compounds underwent epoxide formation followed by internal hydrolysis and also produced a cysteine conjugate.

Conclusion

Human hepatic metabolic profiles were generated for PB-22 and 5F-PB-22. Pentylindole-3-carboxylic acid, hydroxypentyl-PB-22 and PB-22 pentanoic acid for PB-22, and 5′-fluoropentylindole-3-carboxylic acid, PB-22 pentanoic acid and the hydroxy-5F-PB-22 metabolite with oxidation at the quinoline system for 5F-PB-22 are likely the best targets to incorporate into analytical methods for urine to document PB-22 and 5F-PB-22 intake.

Metabolism of synthetic cannabinoids PB-22 and 5F-PB-22 by human hepatocyte incubation and high-resolution mass spectrometry

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Acknowledgments

This research was funded by the Intramural Research Program of the National Institute on Drug Abuse, National Institutes of Health and AB SCIEX.

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Authors and Affiliations

  1. Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, 251 Bayview Boulevard, Baltimore, MD, 21224, USA

    Ariane Wohlfarth, Adarsh S. Gandhi, Karl B. Scheidweiler & Marilyn A. Huestis

  2. AB SCIEX, Redwood City, CA, 94404, USA

    Shaokun Pang

  3. Department of Biotransformation, Bristol-Myers Squibb Research and Development, Princeton, NJ, 08543, USA

    Mingshe Zhu

Authors
  1. Ariane Wohlfarth
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  2. Adarsh S. Gandhi
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  3. Shaokun Pang
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  4. Mingshe Zhu
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  5. Karl B. Scheidweiler
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  6. Marilyn A. Huestis
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Correspondence to Marilyn A. Huestis.

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Wohlfarth, A., Gandhi, A.S., Pang, S. et al. Metabolism of synthetic cannabinoids PB-22 and its 5-fluoro analog, 5F-PB-22, by human hepatocyte incubation and high-resolution mass spectrometry. Anal Bioanal Chem 406, 1763–1780 (2014). https://doi.org/10.1007/s00216-014-7668-0

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