PDF(Pubmed)
Abstract：
Following isotonitazene scheduling in 2019, the availability of alternative 2-benzylbenzimidazole opioids (nitazenes) on the global drug market increased, resulting in many fatalities worldwide. Nitazenes are potent µ-opioid receptor agonists with strong narcotic/analgesic effects, and their concentrations in biological matrices are low, making the detection of metabolite biomarkers of consumption crucial to document use in clinical and forensic settings. However, there is little to no data on the metabolism of the most recently available nitazenes, especially desnitro-analogues. The aim of the research was to assess isotonitazene, metonitazene, etodesnitazene, and metodesnitazene human metabolism and identify specific metabolite biomarkers of consumption. The four analogues were incubated with 10-donor-pooled human hepatocytes, and the incubates were analyzed by liquid chromatography-high-resolution tandem mass spectrometry and data mining with Compound Discoverer (Thermo Scientific); the analysis was supported by in silico metabolite predictions with GLORYx open-access software. Metabolites were identified in postmortem blood and/or urine samples from two metonitazene-positive and three etodesnitazene-positive cases following the same workflow, with and without glucuronide hydrolysis in urine, to confirm in vitro results. Twelve, nine, twenty-two, and ten metabolites were identified for isotonitazene, metonitazene, etodesnitazene, and metodesnitazene, respectively. The main transformations were N-deethylation at the N,N-diethylethanamine side chain, O-dealkylation, and further O-glucuronidation. In vitro and autopsy results were consistent, demonstrating the efficacy of the 10-donor-pooled human hepatocyte model to predict human metabolism. We suggest the parent and the corresponding O-dealkyl- and N-deethyl-O-dealkyl metabolites as biomarkers of exposure in urine after glucuronide hydrolysis, and the corresponding N-deethyl metabolite as additional biomarker in blood.
摘要：
继2019年的等氮烯计划之后，全球药物市场上替代2-苄基苯并咪唑阿片类药物（硝基苯）的可用性增加，导致全世界许多人死亡。硝唑烯是强效的μ-阿片受体激动剂，具有很强的麻醉/镇痛作用，它们在生物基质中的浓度很低，使代谢产物生物标志物的检测对于在临床和法医环境中使用文件至关重要。然而,关于最近可用的硝基苯的代谢几乎没有数据，尤其是去硝基类似物.这项研究的目的是评估异氮烯，甲硝唑,etotesnitazene,和美托硝基苯人代谢，并确定消耗的特定代谢物生物标志物。将四个类似物与10个供体汇集的人肝细胞一起孵育，并用CompoundDiscoverer（ThermoScientific）通过液相色谱-高分辨率串联质谱和数据挖掘对孵育物进行分析；该分析得到了使用GLORYx开放式访问软件进行的计算机代谢物预测的支持。在相同的工作流程下，在来自两个美托那嗪阳性和三个美托那嗪阳性病例的死后血液和/或尿液样本中鉴定出代谢物。在尿液中有或没有葡萄糖醛酸水解，以确认体外结果。十二，九,二十二，并鉴定了十种代谢物的异氮氮烯，甲硝唑,etotesnitazene,和美托硝基苯,分别。主要转化是N-去乙基化，N-二乙基乙胺侧链，O-脱烷基化,和进一步的O-葡糖醛酸化。体外和尸检结果一致，证明了10个供体汇集的人类肝细胞模型预测人类代谢的有效性。我们建议将母体和相应的O-脱烷基-和N-脱乙基-O-脱烷基代谢物作为葡萄糖醛酸水解后尿液中暴露的生物标志物，和相应的N-脱乙基代谢物作为血液中的额外生物标志物。
