title: Metabolism of diosbulbin B in vitro and in vivo in rats: formation of reactive metabolites and human enzymes involved.
abstract: Diosbulbin B (DB), a major constituent of the furano-norditerpenes in Dioscorea bulbifera Linn, exhibits potential antineoplasmic activity and hepatotoxicity. The metabolism and reactive metabolites of DB in vitro (with human and animal liver microsomes) and in vivo in rats were investigated. The human enzymes involved in DB metabolism were identified. DB was first catalyzed into reactive metabolites of 2-butene-1,4-dial derivatives dependent on NADPH and then trapped by Tris base or oxidized to hemiacetal lactones (M12 and M13) in microsomal incubations. Tris base was used as buffer constituent and as a trapping agent for aldehyde. Methoxylamine and glutathione (GSH) were also used as trapping agents. DB metabolism in vivo in rats after oral administration was consistent with that in vitro. The structures of M12 and M13, as well as mono-GSH conjugates of DB (M31), were confirmed by nuclear magnetic resonance spectroscopy of the chemically synthesized products. The bioactivation enzymes of DB were identified as CYP3A4/5, 2C9, and 2C19. CYP3A4 was found to be the primary enzyme using human recombinant cytochrome P450 enzymes, specific inhibitory studies, and a relative activity factor approach for pooled human liver microsomes. Michaelis-Menten constants K(m) and V(max) were determined by the formation of M31. The reactive metabolites may be related to the hepatotoxicity of DB. The gender difference in CYP3A expression in mice and rats contributed to the gender-related liver injury and pharmacokinetics in mice and rats, respectively.
authors: Ministry of Education Key Laboratory for Standardization of Chinese Medicines, State Administration of Traditional Chinese Medicine Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China (B.Y., W.L., K.C., Z.W., C.W.); and Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai, China (K.C., Z.W., C.W.).
authors: Baohua
authors: B
authors: Yang
authors: Ministry of Education Key Laboratory for Standardization of Chinese Medicines, State Administration of Traditional Chinese Medicine Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China (B.Y., W.L., K.C., Z.W., C.W.); and Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai, China (K.C., Z.W., C.W.).
authors: Wei
authors: W
authors: Liu
authors: Ministry of Education Key Laboratory for Standardization of Chinese Medicines, State Administration of Traditional Chinese Medicine Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China (B.Y., W.L., K.C., Z.W., C.W.); and Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai, China (K.C., Z.W., C.W.).
authors: Kaixian
authors: K
authors: Chen
authors: Ministry of Education Key Laboratory for Standardization of Chinese Medicines, State Administration of Traditional Chinese Medicine Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China (B.Y., W.L., K.C., Z.W., C.W.); and Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai, China (K.C., Z.W., C.W.) wchcxm@hotmail.com wangzht@hotmail.com.
authors: Zhengtao
authors: Z
authors: Wang
authors: Ministry of Education Key Laboratory for Standardization of Chinese Medicines, State Administration of Traditional Chinese Medicine Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China (B.Y., W.L., K.C., Z.W., C.W.); and Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai, China (K.C., Z.W., C.W.) wchcxm@hotmail.com wangzht@hotmail.com.
authors: Changhong
authors: C
authors: Wang
mesh_terms: D065767:Activation, Metabolic; D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006168:Guinea Pigs; D006576:Heterocyclic Compounds, 4 or More Rings / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism; D015394:Molecular Structure; D011817:Rabbits; D051381:Rats
chemical_list: D006576:Heterocyclic Compounds, 4 or More Rings; C569454:diosbulbin B; D003577:Cytochrome P-450 Enzyme System