Han Lingyu, Wang Hongjie, Si Nan, Ren Wei, Gao Bo, Li Yan, Yang Jian, Xu Miao, Zhao Haiyu, Bian Baolin
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Anhui Jinchan Biochemistry Company Ltd., Huaibei, 235000, China.
Anal Bioanal Chem. 2016 Apr;408(10):2485-95. doi: 10.1007/s00216-016-9345-y. Epub 2016 Feb 11.
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.
蟾蜍二烯羟酸内酯是一类多羟基甾体化合物，具有显著的抗肿瘤活性。在本研究中，采用超高效液相色谱（UHPLC）结合LTQ Orbitrap质谱仪对10种蟾蜍二烯羟酸内酯的39种代谢产物进行了检测和鉴定。结果表明，羟基化和脱氢是蟾蜍二烯羟酸内酯在人肝微粒体（HLMs）中的主要代谢途径。发现CYP3A4是主要的代谢酶，而CYP2D6仅介导脱氢反应。建立了一种系统的、经过验证的等当量蟾蜍二烯羟酸内酯代谢产物细胞毒性评价方法。选择海葱苷元（1）、海葱醇（2）、华蟾酥毒基（3）、蟾毒灵（5）和蟾酥灵（6），测定它们在HLMs中孵育前后对HepG2细胞的细胞毒性。所有测试样品均通过经过验证的固相萃取（SPE）方法进行富集。尽管代谢产物的细胞毒性在不同程度上比母体化合物弱，但其作用仍然很强。
