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In vitro and in vivo human metabolism of a new synthetic cannabinoid NM-2201 (CBL-2201)

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Abstract

In 2014, NM-2201 (CBL-2201), a novel synthetic cannabinoid (SC), was detected by scientists at Russian and US laboratories. It has been already added to the list of scheduled drugs in Japan, Sweden and Germany. Unfortunately, no human metabolism data are currently available, which makes it challenging to confirm its intake, especially given that all SCs investigated thus far have been found to be extensively metabolized. The present study aims to recommend appropriate marker metabolites by investigating NM-2201 metabolism in human hepatocytes, and to confirm the results in authentic human urine specimens. For the metabolic stability assay, 1 µM NM-2201 was incubated in human liver microsomes (HLMs) for up to 1 h; for metabolite profiling, 10 µM of NM-2201 was incubated in human hepatocytes for 3 h. Two authentic urine specimens from NM-2201-positive cases were subjected to β-glucuronidase hydrolysis prior to analysis. The identification of metabolites in hepatocyte samples and urine specimens was achieved with high-resolution mass spectrometry via information-dependent acquisition. NM-2201 was quickly metabolized in HLMs, with an 8.0-min half-life. In human hepatocyte incubation samples, a total of 13 NM-2201 metabolites were identified, generated mainly from ester hydrolysis and further hydroxylation, oxidative defluorination and subsequent glucuronidation. M13 (5-fluoro PB-22 3-carboxyindole) was found to be the major metabolite. In the urine specimens, the parent drug NM-2201 was not detected; M13 was the predominant metabolite after β-glucuronidase hydrolysis. Therefore, based on the results of our study, we recommend M13 as a suitable urinary marker metabolite for confirming NM-2201 and/or 5F-PB-22 intake.

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Acknowledgments

This research is supported by the Intramural Research Program of the National Institute on Drug Abuse, National Institutes of Health. NM-2201 was generously donated by the US Drug Enforcement Administration. We also appreciate help from Dr. Ariane Wohlfarth in performing the HLMs metabolic stability assays.

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Authors and Affiliations

  1. Chemistry and Drug Metabolism Section, Clinical Pharmacology and Therapeutics Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, 251 Bayview Blvd, Suite 200 Room 05A727, Baltimore, MD, 21224, USA

    Xingxing Diao, Jeremy Carlier, Karl B. Scheidweiler & Marilyn A. Huestis

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

    Mingshe Zhu

  3. SCIEX, Redwood City, CA, 94065, USA

    Shaokun Pang

  4. Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, 58758, Linköping, Sweden

    Robert Kronstrand

  5. Department of Drug Research, University of Linköping, 58185, Linköping, Sweden

    Robert Kronstrand

  6. University of Maryland School of Medicine, Baltimore, MD, 21224, USA

    Marilyn A. Huestis

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  1. Xingxing Diao
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  2. Jeremy Carlier
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  4. Shaokun Pang
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Correspondence to Karl B. Scheidweiler.

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Diao, X., Carlier, J., Zhu, M. et al. In vitro and in vivo human metabolism of a new synthetic cannabinoid NM-2201 (CBL-2201). Forensic Toxicol 35, 20–32 (2017). https://doi.org/10.1007/s11419-016-0326-9

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