title: Metabolites profiling of 10 bufadienolides in human liver microsomes and their cytotoxicity variation in HepG2 cell.
abstract: Bufadienolides, a class of polyhydroxy steroids, exhibit significant antitumor activity. In this study, a total of 39 metabolites from 10 bufadienolides were detected and identified by ultrahigh-performance liquid chromatography (UHPLC) coupled with an LTQ Orbitrap mass spectrometer. The results showed that hydroxylation and dehydrogenation were the major metabolic pathways of bufadienolides in human liver microsomes (HLMs). CYP3A4 was found to be the major metabolic enzyme and CYP2D6 only mediated the dehydrogenation reaction. A systematic validated cytotoxicity evaluation method for bufadienolide metabolites at equal equivalents was established. Hellebrigenin (1), hellebrigenol (2), arenobufagin (3), bufotalin (5), and bufalin (6) were selected to determine their cytotoxicity against HepG2 cells before and after incubation in HLMs. All the test samples were enriched by a validated solid-phase extraction (SPE) method. Although the cytotoxicities of metabolites were weaker than those of the parent compounds to different degrees, their effects were still strong.
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors: Lingyu
authors: L
authors: Han
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors: Hongjie
authors: H
authors: Wang
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors: Nan
authors: N
authors: Si
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors: Wei
authors: W
authors: Ren
authors: Anhui Jinchan Biochemistry Company Ltd., Huaibei, 235000, China.
authors: Bo
authors: B
authors: Gao
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors: Yan
authors: Y
authors: Li
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors: Jian
authors: J
authors: Yang
authors: State Intellectual Property Office of the P.R.C, Beijing, 100088, China.
authors: Miao
authors: M
authors: Xu
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China. hyzhao@icmm.ac.cn.
authors: Haiyu
authors: H
authors: Zhao
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China. blbian@icmm.ac.cn.
authors: Baolin
authors: B
authors: Bian
mesh_terms: D002018:Bufanolides / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D056945:Hep G2 Cells; D006801:Humans; D008862:Microsomes, Liver / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D002018:Bufanolides; C087925:bufadienolide
keywords: Bufadienolides; CYP2D6; CYP3A4; HepG2 cell line; Metabolites