School of investigation, People's Public Security University of China, 1st Muxidi South Lane, Xicheng District, Beijing, 100038, China.
The Criminal Investigation Department of Beijing Public Security Bureau (Key Laboratory of Forensic Toxicology, Ministry of Public Security), 1st Longgang Road, Haidian District, Beijing, 100085, China.
J Pharm Biomed Anal. 2022 Oct 25;220:114985. doi: 10.1016/j.jpba.2022.114985. Epub 2022 Aug 8.
MDMB-4en-PINACA (Methyl 3,3-dimethyl-2-[1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido] butanoate) is a potent agonist of the CB1 receptor. In 2021, it was one of the most common synthetic cannabinoid receptor agonists (SCRAs) seized by the Beijing Drug Control Agency. MDMB-4en-PINACA can be hard to detect in biological specimens because of ester hydrolysis. In this work, a highly sensitive liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method was developed for the detection of MDMB-4en-PINACA metabolites in urine, serum, and hair samples. Metabolites from authentic samples were compared with those from human liver microsomes (HLMs) in vitro and in zebrafish in vivo. A total of 75 metabolites, including 44 previously unreported metabolites, were identified from urine samples. We found that 11 metabolic pathways were involved in MDMB-4en-PINACA metabolism, including acetylation, a novel metabolic pathway for SCRAs. Our results revealed that ester hydrolysis and hydroxylation were to the major metabolic pathways involved in MDMB-4en-PINACA metabolism. Using serum samples, we detected 9 metabolites along with the parent drug. Only the parent drug was detected using hair samples. The existence of ADB-4en-PINACA makes the currently used biomarkers for MDMB-4enPINACA not very specific for the intake of MDMB-4en-PINACA. Therefore, based on the identified metabolites and their structural features, we propose more sensitive screening tactics for MDMB-4en-PINACA using urine and serum samples.
MDMB-4en-PINACA（3,3-二甲基-2-[1-(戊-4-烯-1-基)-1H-吲哚-3-甲酰胺基]丁酸甲酯）是一种有效的 CB1 受体激动剂。2021 年，它是北京药品监管机构缴获的最常见的合成大麻素受体激动剂（SCRAs）之一。由于酯水解，MDMB-4en-PINACA 很难在生物样本中检测到。在这项工作中，开发了一种高灵敏度的液相色谱-高分辨质谱（LC-HRMS）方法，用于检测尿液、血清和头发样本中的 MDMB-4en-PINACA 代谢物。从真实样本中鉴定的代谢物与体外人肝微粒体（HLMs）和体内斑马鱼中的代谢物进行了比较。从尿液样本中鉴定出 75 种代谢物，包括 44 种以前未报道过的代谢物。我们发现，MDMB-4en-PINACA 代谢涉及 11 种代谢途径，包括乙酰化，这是一种新的 SCRAs 代谢途径。我们的研究结果表明，酯水解和羟化是 MDMB-4en-PINACA 代谢的主要代谢途径。使用血清样本，我们检测到了 9 种代谢物和母体药物。仅使用毛发样本检测到母体药物。ADB-4en-PINACA 的存在使得目前用于 MDMB-4en-PINACA 的生物标志物对 MDMB-4en-PINACA 的摄入不太特异。因此，基于鉴定的代谢物及其结构特征，我们提出了使用尿液和血清样本更敏感的 MDMB-4en-PINACA 筛选策略。
