title: The Effect of Oxidation on Berberine-Mediated CYP1 Inhibition: Oxidation Behavior and Metabolite-Mediated Inhibition.
abstract: The protoberberine alkaloid berberine carries methylenedioxy moiety and exerts a variety of pharmacological effects, such as anti-inflammation and lipid-lowering effects. Berberine causes potent CYP1B1 inhibition, whereas CYP1A2 shows resistance to the inhibition. To reveal the influence of oxidative metabolism on CYP1 inhibition by berberine, berberine oxidation and the metabolite-mediated inhibition were determined. After NADPH-fortified preincubation of berberine with P450, the inhibition of CYP1A1 and CYP1B1 variants (CYP1B1.1, CYP1B1.3, and CYP1B1.4) by berberine was not enhanced, and CYP1A2 remained resistant. Demethyleneberberine was identified as the most abundant metabolite of CYP1A1- and CYP1B1-catalyzed oxidations, and thalifendine was generated at a relatively low rate. CYP1A1-catalyzed berberine oxidation had the highest maximal velocity (V max) and exhibited positive cooperativity, suggesting the assistance of substrate binding when the first substrate was present. In contrast, the demethylenation by CYP1B1 showed the property of substrate inhibition. CYP1B1-catalyzed berberine oxidation had low K m values, but it had V max values less than 8% of those of CYP1A1. The dissociation constants generated from the binding spectrum and fluorescence quenching suggested that the low K m values of CYP1B1-catalyzed oxidation might include more than the rate constants describing berberine binding. The natural protoberberine/berberine fmetabolites with methylenedioxy ring-opening (palmatine, jatrorrhizine, and demethyleneberberine) and the demethylation (thalifendine and berberrubine) caused weak CYP1 inhibition. These results demonstrated that berberine was not efficiently oxidized by CYP1B1, and metabolism-dependent irreversible inactivation was minimal. Metabolites of berberine caused a relatively weak inhibition of CYP1.
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.).
authors.forename: Sheng-Nan
authors.initials: SN
authors.lastname: Lo
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.).
authors.forename: Chien-Chang
authors.initials: CC
authors.lastname: Shen
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.).
authors.forename: Chia-Yu
authors.initials: CY
authors.lastname: Chang
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.).
authors.forename: Keng-Chang
authors.initials: KC
authors.lastname: Tsai
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.).
authors.forename: Chiung-Chiao
authors.initials: CC
authors.lastname: Huang
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.).
authors.forename: Tian-Shung
authors.initials: TS
authors.lastname: Wu
authors.affiliation: Divisions of Basic Chinese Medicine (S.-N.L., C.-Y.C., C.-C.H., Y.-F.U.), Chinese Medicinal Chemistry (C.-C.S.), and Chinese Materia Medica Development (K.-C.T.), National Research Institute of Chinese Medicine, Taipei, Taiwan, Republic of China; Institute of Biopharmaceutical Sciences, School of Life Science (S.-N.L., Y.-F.U.) and Department of Pharmacology, School of Medicine (Y.-F.U.), National Yang-Ming University, Taipei, Taiwan, Republic of China; Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, Republic of China (C.-Y.C., Y.-F.U.); and Department of Chemistry, National Chung-Kung University, Tainan, Taiwan, Republic of China (T.-S.W.) ueng@nricm.edu.tw.
authors.forename: Yune-Fang
authors.initials: YF
authors.lastname: Ueng
mesh_terms: D001599:Berberine / Q000031:analogs & derivatives / Q000378:metabolism / Q000493:pharmacokinetics* / Q000494:pharmacology*; D001711:Biotransformation; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism*; D065633:Cytochrome P-450 CYP1B1 / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000494:pharmacology*; D003640:Dealkylation; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D009249:NADP / Q000494:pharmacology; D010084:Oxidation-Reduction
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D007527:Isoenzymes; D001599:Berberine; C477156:thalifendine; D009249:NADP; C585496:CYP1B1 protein, human; D019363:Cytochrome P-450 CYP1A1; D019388:Cytochrome P-450 CYP1A2; D065633:Cytochrome P-450 CYP1B1