Erratico Claudio, Negreira Noelia, Norouzizadeh Helia, Covaci Adrian, Neels Hugo, Maudens Kristof, van Nuijs Alexander L N
Toxicological Center, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk-Antwerp, Belgium.
Drug Test Anal. 2015 Oct;7(10):866-76. doi: 10.1002/dta.1796. Epub 2015 Apr 12.
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.
N-[(1S)-1-(氨基甲酰基)-2-甲基丙基]-1-(环己基甲基)-1H-吲唑-3-甲酰胺（AB-CHMINACA）是一种最近引入的合成大麻素。目前，尚无关于AB-CHMINACA在人体体外或体内代谢的信息。因此，用于筛查AB-CHMINACA使用情况的生物监测研究缺乏关于潜在生物标志物（如代谢物）的任何信息。为了填补这一空白，我们使用人肝微粒体（HLM）研究了AB-CHMINACA的体外代谢。使用液相色谱-飞行时间质谱法监测AB-CHMINACA代谢物的形成。检测到AB-CHMINACA的26种代谢物，包括7种单羟基化和6种二羟基化代谢物以及一种由AB-CHMINACA的N-脱烷基化产生的代谢物，所有这些均由细胞色素P450（CYP）酶产生。还形成了两种可能由酰胺酶产生的羧化代谢物和五种葡萄糖醛酸化代谢物。五种单羟基化和一种羧化代谢物可能是检测到的主要代谢物。使用一组七种人重组CYP（rCYP）测试了个体CYP在AB-CHMINACA代谢物形成中的作用。HLM产生的所有羟基化AB-CHMINACA代谢物也由测试的rCYP产生，其中rCYP3A4是最活跃的酶。AB-CHMINACA的大多数体外代谢物也存在于一名AB-CHMINACA使用者的尿液中，因此表明使用体外代谢实验预测AB-CHMINACA体内代谢所获得结果的可靠性。现在已经可靠地鉴定出在使用尿液样本的生物监测研究中要靶向的AB-CHMINACA代谢物，可用于常规分析。
