Centre for Forensic Science, School of Mathematical and Physical Sciences, University of Technology Sydney (UTS), PO Box 123, Broadway, NSW, 2007, Australia.
Climate Change Cluster, University of Technology Sydney (UTS), PO Box 123, Broadway, NSW, 2007, Australia.
AAPS J. 2018 Mar 8;20(2):42. doi: 10.1208/s12248-018-0209-6.
The number of new psychoactive substances keeps on rising despite the controlling efforts by law enforcement. Although metabolism of the newly emerging drugs is continuously studied to keep up with the new additions, the exact structures of the metabolites are often not identified due to the insufficient sample quantities for techniques such as nuclear magnetic resonance (NMR) spectroscopy. The aim of the study was to characterise several metabolites of the synthetic cannabinoid (1-pentyl-1H-indol-3-yl) (2,2,3,3-tetramethylcyclopropyl) methanone (UR-144) by NMR spectroscopy after the incubation with the fungus Cunninghamella elegans. UR-144 was incubated with C. elegans for 72 h, and the resulting metabolites were chromatographically separated. Six fractions were collected and analysed by NMR spectroscopy. UR-144 was also incubated with human liver microsomes (HLM), and the liquid chromatography-high resolution mass spectrometry analysis was performed on the HLM metabolites with the characterised fungal metabolites as reference standards. Ten metabolites were characterised by NMR analysis including dihydroxy metabolites, carboxy and hydroxy metabolites, a hydroxy and ketone metabolite, and a carboxy and ketone metabolite. Of these metabolites, dihydroxy metabolite, carboxy and hydroxy metabolites, and a hydroxy and ketone metabolite were identified in HLM incubation. The results indicate that the fungus is capable of producing human-relevant metabolites including the exact isomers. The capacity of the fungus C. elegans to allow for NMR structural characterisation by enabling production of large amounts of metabolites makes it an ideal model to complement metabolism studies.
尽管执法部门在努力控制，但新精神活性物质的数量仍在不断增加。尽管新兴药物的代谢不断得到研究，以跟上新的添加物，但由于技术（如核磁共振（NMR）光谱法）所需的样本数量不足，通常无法确定代谢物的确切结构。本研究的目的是通过核磁共振（NMR）光谱法对合成大麻素（1-戊基-1H-吲哚-3-基）（2,2,3,3-四甲基环丙基）甲酮（UR-144）与真菌 Cunninghamella elegans 孵育后的几种代谢物进行表征。将 UR-144 与 C. elegans 孵育 72 小时，然后通过色谱法分离得到的代谢物。收集了六个馏分并通过 NMR 光谱法进行了分析。还将 UR-144 与人肝微粒体（HLM）孵育，并使用作为参考标准的特征化真菌代谢物对 HLM 代谢物进行液相色谱-高分辨率质谱分析。通过 NMR 分析鉴定了包括二羟基代谢物、羧基和羟基代谢物、羟基和酮代谢物以及羧基和酮代谢物在内的十种代谢物。其中，在 HLM 孵育中鉴定出二羟基代谢物、羧基和羟基代谢物以及羟基和酮代谢物。结果表明，真菌能够产生包括立体异构体在内的与人相关的代谢物。真菌 C. elegans 能够产生大量代谢物，从而能够进行 NMR 结构表征，使其成为补充代谢研究的理想模型。
