Characterization of the in Vitro Metabolic Profile of Evodiamine in Human Liver Microsomes and Hepatocytes by UHPLC-Q Exactive Mass Spectrometer

Abstract

Evodiamine is an indoloquinazoline alkaloid isolated from the fruit of Evodia rutaecarpa, which has a wide range of pharmacological effects like anti-tumor and anti-inflammatory effects. This study was intended to investigate the metabolic characteristics of evodiamine in human liver microsomes and hepatocytes by ultra-high performance liquid chromatography coupled with a Q Exactive mass spectrometer. A total of 12 phase I metabolites were detected in human liver microsomes; whereas in human hepatocytes 19 metabolites, including seven phase II metabolites were detected. The structures of the metabolites were characterized based on their accurate masses, fragment ions, and chromatographic retention times. Four metabolites (M1, M2, M5, and M7) were further unambiguously confirmed by matching their retention times, accurate masses, and fragment ions with those of their reference standards. Among these metabolites, 12 metabolites are first identified (M2, M5–M8, M10–M13, and M17–M19). The current study revealed that oxygenation, N-demethylation, dehydrogenation, glucuronidation, and GSH conjugation were the major metabolic pathways for evodiamine. This study elucidated the detailed metabolite profiles of evodiamine, which is helpful in predicting in vivo metabolism of evodiamine in human and in understanding the elimination mechanism of evodiamine and in turn, the effectiveness and toxicity.

Research progress on evodiamine, a bioactive alkaloid of Evodiae fructus: Focus on its anti‑cancer activity and bioavailability (Review)

“…Therefore, the metabolic bioactivation of EVO may be closely associated with its pharmacological effects and toxicity (35). Aromatic, aliphatic hydroxylation, N-demethylation, oxygenation, dehydrogenation, glucuronidation and GSH conjugation are involved in the metabolic pathways of EVO, where a methyl group in position 14 of EVO may contribute to its metabolic properties (87,88). In vitro incubation of EVO with human and rat liver microsomes, in the presence of NADPH, resulted in the formation of four mono-hydroxylated metabolites and one N-demethylated metabolite (87).…”

“…Several GSH conjugates of EVO were also found after the addition of sulfhydryl nucleophiles to EVO (34). Recently, a total of 12 phase I metabolites of EVO were found in human liver microsomes, whilst 12 phase I metabolites and seven phase II conjugated metabolites were identified and quantified in human hepatocytes (88).…”

Design, Synthesis, and Biological Evaluation of Novel Evodiamine Derivatives as Potential Antihepatocellular Carcinoma Agents

“…Natural products and their synthetic derivatives have always been important sources of active molecules for anticancer drugs. − Evodia rutaecarpa, also known as Wu-Chu-Yu, is a traditional Chinese medicine widely applied to headaches, diarrhea, and postpartum bleeding. − Evodiamine (EVO), a quinazolinocarboline alkaloid, is isolated from Evodiae fructus, which elicits a variety of biological activities covering anti-Alzheimer’s disease, anti-inflammatory, anticardiovascular disease, and antitumor. − Particularly, EVO and its derivatives can play an anticancer role by double inhibition of topoisomerase I and II (Top1 and Top2), which are highly expressed in cancer tissues. − Nevertheless, its further application in anti-HCC is limited because of its low activity and easing to metabolize into inactive products. , Additionally, the research on EVO and its derivatives in liver cancer is scarce. Given this, structure modification of EVO is of great significance to develop novel efficient anti-HCC agents.…”

“…24−31 application in anti-HCC is limited because of its low activity and easing to metabolize into inactive products. 32,33 Additionally, the research on EVO and its derivatives in liver cancer is scarce. Given this, structure modification of EVO is of great significance to develop novel efficient anti-HCC agents.…”

A Systematic Strategy for the Characterization of 2,3,5,4′-Tetrahydroxystilbene-2-O-β-d-glucoside Metabolites In Vivo by Ultrahigh Performance Liquid Chromatography Coupled with a Q Exactive-Orbitrap Mass System

“…In recent years, because of the development of high resolution and high sensitivity of the mass spectrometry technology, liquid chromatography coupled with mass spectrometer systems have been used as effective and reliable analytical instruments to identify metabolites. 7,8 The Thermo Q Exactive mass spectrometer combines a high-performance In this article, a novel strategy based on the UHPLC-Q Exactive-Orbitrap MS system and its data-processing tools of MDF, XIC, DBS, PIF, and targeted screening were developed to quickly screen and systematically identify metabolites of THSG in vivo. A total of 75 metabolites were detected in rat plasma, bile, urine, and tissues (heart, liver, spleen, lung, kidney, and stomach), and the probable metabolic pathways of THSG were proposed.…”