title: In vitro and in vivo human metabolism of the synthetic cannabinoid AB-CHMINACA.
abstract: N-[(1S)-1-(aminocarbonyl)-2-methylpropyl]-1-(cyclohexylmethyl)-1H-indazole-3-carboxamide (AB-CHMINACA) is a recently introduced synthetic cannabinoid. At present, no information is available about in vitro or in vivo human metabolism of AB-CHMINACA. Therefore, biomonitoring studies to screen AB-CHMINACA consumption lack any information about the potential biomarkers (e.g. metabolites) to target. To bridge this gap, we investigated the in vitro metabolism of AB-CHMINACA using human liver microsomes (HLMs). Formation of AB-CHMINACA metabolites was monitored using liquid chromatography coupled to time-of-flight mass spectrometry. Twenty-six metabolites of AB-CHMINACA were detected including seven mono-hydroxylated and six di-hydroxylated metabolites and a metabolite resulting from N-dealkylation of AB-CHMINACA, all produced by cytochrome P450 (CYP) enzymes. Two carboxylated metabolites, likely produced by amidase enzymes, and five glucuronidated metabolites were also formed. Five mono-hydroxylated and one carboxylated metabolite were likely the major metabolites detected. The involvement of individual CYPs in the formation of AB-CHMINACA metabolites was tested using a panel of seven human recombinant CYPs (rCYPs). All the hydroxylated AB-CHMINACA metabolites produced by HLMs were also produced by the rCYPs tested, among which rCYP3A4 was the most active enzyme. Most of the in vitro metabolites of AB-CHMINACA were also present in urine obtained from an AB-CHMINACA user, therefore showing the reliability of the results obtained using the in vitro metabolism experiments conducted to predict AB-CHMINACA in vivo metabolism. The AB-CHMINACA metabolites to target in biomonitoring studies using urine samples are now reliably identified and can be used for routine analysis.
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Claudio
authors: C
authors: Erratico
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Noelia
authors: N
authors: Negreira
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Helia
authors: H
authors: Norouzizadeh
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Adrian
authors: A
authors: Covaci
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Hugo
authors: H
authors: Neels
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Kristof
authors: K
authors: Maudens
authors: Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
authors: Alexander L N
authors: AL
authors: van Nuijs
mesh_terms: D002186:Cannabinoids / Q000737:chemistry / Q000378:metabolism*; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D015198:Designer Drugs / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D007191:Indazoles / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000378:metabolism*; D014633:Valine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: D002186:Cannabinoids; D015198:Designer Drugs; D007191:Indazoles; D003577:Cytochrome P-450 Enzyme System; D014633:Valine; C000606289:N-(1-(aminocarbonyl)-2-methylpropyl)-1-(cyclohexylmethyl)-1H-indazole-3-carboxamide
keywords: AB-CHMINACA; LC-QTOF/MS; human liver microsomes; in vitro metabolism