High-Resolution Mass Spectrometry for Characterizing the Metabolism of Synthetic Cannabinoid THJ-018 and Its 5-Fluoro Analog THJ-2201 after Incubation in Human Hepatocytes

Abstract

BACKGROUND Despite increasing prevalence of novel psychoactive substances, no human metabolism data are currently available, complicating laboratory documentation of intake in urine samples and assessment of the drugs' pharmacodynamic, pharmacokinetic, and toxicological properties. In 2014, THJ-018 and THJ-2201, synthetic cannabinoid indazole analogs of JWH-018 and AM-2201, were identified, with the National Forensic Laboratory Information System containing 220 THJ-2201 reports. Because of numerous adverse events, the Drug Enforcement Administration listed THJ-2201 as Schedule I in January 2015. METHODS We used high-resolution mass spectrometry (HR-MS) (TripleTOF 5600+) to identify optimal metabolite markers after incubating 10 μmol/L THJ-018 and THJ-2201 in human hepatocytes for 3 h. Data were acquired via full scan and information-dependent acquisition triggered product ion scans with mass defect filter. In silico metabolite predictions were performed with MetaSite and compared with metabolites identified in human hepatocytes. RESULTS Thirteen THJ-018 metabolites were detected, with the major metabolic pathways being hydroxylation on the N-pentyl chain and further oxidation or glucuronidation. For THJ-2201, 27 metabolites were observed, predominantly oxidative defluorination plus subsequent carboxylation or glucuronidation, and glucuronidation of hydroxylated metabolites. Dihydrodiol formation on the naphthalene moiety was observed for both compounds. MetaSite prediction matched well with THJ-018 hepatocyte metabolites but underestimated THJ-2201 oxidative defluorination. CONCLUSIONS With HR-MS for data acquisition and processing, we characterized THJ-018 and THJ-2201 metabolism in human hepatocytes and suggest appropriate markers for laboratories to identify THJ-018 and THJ-2201 intake and link observed adverse events to these new synthetic cannabinoids.

In Vitro and In Vivo Human Metabolism of Synthetic Cannabinoids FDU-PB-22 and FUB-PB-22

“…Oxidative defluorination was the major metabolic pathway for ω-fluoropentyl chain containing drugs, such as 5F-PB-22, 5F-AKB-48, and THJ-2201 (8,29,30). In our study, the pfluorobenzyl moiety was metabolically stable in the hepatocyte incubation and in vivo, with no biotransformation identified.…”

Metabolic Profile of Synthetic Cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans

“…The only fragment ion detected was at m/z 230 suggesting a quinoline moiety with dihydrodiol formation and a pentylindole moiety with a hydroxy group. Despite the lack of the m/z 144 fragment ion, the position of the hydroxy group can be considered to be at the terminal carbon of the pentyl side chain since oxidative defluorination is a commonly reported pathway to simultaneously defluorinate and hydroxylate fluorinated synthetic cannabinoids [2,6,7,16,18,19] and it has been reported for C. elegans [33].…”

In Vitro, in Vivo and In Silico Metabolic Profiling of α-Pyrrolidinopentiothiophenone, A Novel Thiophene Stimulant

“…In order to generate the most comprehensive metabolic profile of a substance, in vitro human hepatocyte incubations, followed by analysis with liquid chromatography coupled to high-resolution MS (LC-HRMS) and softwareassisted data mining were proven to be a promising approach [30][31][32]. Human hepatocytes offer a more complete metabolic profile than HLM since hepatocytes contain all hepatic enzymes involved in drug metabolism (both phase I and II transformations) as well as endogenous cofactors, provide the natural orientation of membrane enzymes and produce metabolites in concentrations similar to in vivo [33].…”