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First Characterization of AKB-48 Metabolism, a Novel Synthetic Cannabinoid, Using Human Hepatocytes and High-Resolution Mass Spectrometry

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Abstract

Since the federal authorities scheduled the first synthetic cannabinoids, JWH-018 and JWH-073, new synthetic cannabinoids were robustly marketed. N-(1-Adamantyl)-1-pentylindazole-3-carboxamide (AKB-48), also known as APINACA, was recently observed in Japanese herbal smoking blends. The National Forensic Laboratory Information System registered 443 reports of AKB-48 cases in the USA from March 2010 to January 2013. In May 2013, the Drug Enforcement Administration listed AKB-48 as a Schedule I drug. Recently, AKB-48 was shown to have twice the CB1 receptor binding affinity than CB2. These pharmacological effects and the difficulty in detecting the parent compound in urine highlight the importance of metabolite identification for developing analytical methods for clinical and forensic investigations. Using human hepatocytes and TripleTOF mass spectrometry, we identified 17 novel phase I and II AKB-48 metabolites, products of monohydroxylation, dihydroxylation, or trihydroxylation on the aliphatic adamantane ring or N-pentyl side chain. Glucuronide conjugation of some mono- and dihydroxylated metabolites also occurred. Oxidation and dihydroxylation on the adamantane ring and N-pentyl side chain formed a ketone. More metabolites were identified after 3 h of incubation than at 1 h. For the first time, we present a AKB-48 metabolic scheme obtained from human hepatocytes and high-resolution mass spectrometry. These data are needed to develop analytical methods to identify AKB-48 consumption in clinical and forensic testing.

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Abbreviations

AKB-48:

N-(1-Adamantyl)-1-pentylindazole-3-carboxamide

CB:

Cannabinoid

cps:

Counts per second

CYP:

Cytochrome P450

IDA:

Information-dependent acquisition

LC-MS:

Liquid chromatography mass spectrometry

MDF:

Mass defect filter

MW:

Molecular weight

NFLIS:

National Forensic Laboratory Information System

THC:

Delta 9-tetrahydrocannabinol

TOF:

Time of flight

UDPGT:

Uridine diphosphate glucuronosyltransferase

XICs:

Extracted ion chromatograms

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Acknowledgments

This research was supported by the Intramural Research Program of the National Institute on Drug Abuse, National Institutes of Health.

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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, Suite 200, Room 05A-721, Baltimore, Maryland, 21224, USA

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

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

    Mingshe Zhu

  3. AB SCIEX, Foster City, California, 94404, USA

    Shaokun Pang & Hua-fen Liu

Authors
  1. Adarsh S. Gandhi
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  2. Mingshe Zhu
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  3. Shaokun Pang
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  4. Ariane Wohlfarth
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  5. Karl B. Scheidweiler
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  6. Hua-fen Liu
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  7. Marilyn A. Huestis
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Corresponding author

Correspondence to Marilyn A. Huestis.

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Gandhi, A.S., Zhu, M., Pang, S. et al. First Characterization of AKB-48 Metabolism, a Novel Synthetic Cannabinoid, Using Human Hepatocytes and High-Resolution Mass Spectrometry. AAPS J 15, 1091–1098 (2013). https://doi.org/10.1208/s12248-013-9516-0

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  • DOI: https://doi.org/10.1208/s12248-013-9516-0

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