Liquid chromatography/tandem mass spectrometric determination of inhibition of human cytochrome P450 isozymes by resveratrol and resveratrol-3-sulfate.
Liquid chromatography/tandem mass spectrometric determination of inhibition of human cytochrome P450 isozymes by resveratrol and resveratrol-3-sulfate.
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DOI：
10.1002/rcm.918
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发表时间：
2003-02
期刊：
影响因子：
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通讯作者：
Chongwoo Yu;Y. Shin;J. Kosmeder;J. Pezzuto;R. V. van Breemen
中科院分区：
文献类型：
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作者：
Chongwoo Yu;Y. Shin;J. Kosmeder;J. Pezzuto;R. V. van Breemen
反式白藜芦醇是一种存在于葡萄、葡萄酒、花生和小红莓中的酚类植物保留素，已被报道既具有抗癌、抗氧化、植物雌激素和心脏保护活性，又是细胞色素P450(CYP)3A4的微弱抑制剂，可能在药物相互作用中具有重要意义。由于反式白藜芦醇在体内可以快速转化为主要的反式白藜芦醇-3-硫酸酯，因此建立了一种快速、选择性和灵敏的LC/MS/MS方法来研究反式白藜芦醇-3-硫酸酯对人细胞色素P450的抑制作用。利用表达的人重组同工酶，研究了反式白藜芦醇和反式白藜芦醇-3-硫酸酯对细胞色素P450选择性底物(细胞色素P450底物细胞色素P450的代谢)的影响。为了验证方法的有效性，采用LC/MS/MS检测了不同浓度(0-50微米)的已知选择性抑制剂对多种细胞色素P450同工酶的抑制作用。标准同工酶抑制剂的IC(50)值分别为3.2、1.4、8.9、0.2和0.3微米，与文献值一致。反式白藜芦醇对CYP2C19的IC(50)值为11.6微米，对CYP3A4的IC(50)值为1.1微米，而对所有其他CYP同工酶的IC(50)值均超过50微米，表明没有抑制作用。反式-白藜芦醇-3-硫酸酯无酶抑制作用。我们的结果表明，反式白藜芦醇是细胞色素P450同工酶的抑制剂，但其主要代谢产物反式白藜芦醇-3-硫酸酯不是所研究的任何细胞色素P450同工酶的抑制剂。
trans-Resveratrol, a phenolic phytoalexin occurring in grapes, wine, peanuts, and cranberries, has been reported to both have anticarcinogenic, antioxidative, phytoestrogenic, and cardioprotective activities, and to be a weak inhibitor of cytochrome P450 (CYP)3A4, which might have significance for drug-drug interactions. Since trans-resveratrol is rapidly converted in vivo to primarily trans-resveratrol-3-sulfate, a rapid, selective, and sensitive method using liquid chromatography/tandem mass spectrometry (LC/MS/MS) was developed to investigate human cytochrome P450 inhibition by trans-resveratrol-3-sulfate. Effects of trans-resveratrol and trans-resveratrol-3-sulfate on the metabolism of selective cytochrome P450 substrates (CYP1A2/ethoxyresorufin, CYP2C9/diclofenac, CYP2C19/(S)-mephenytoin, CYP2D6/bufuralol, CYP3A4/testosterone) were monitored using cDNA-expressed human recombinant isozymes. For method validation, LC/MS/MS was used to measure the inhibition of various cytochrome P450 isozymes by different concentrations (0-50 microM) of known selective inhibitors. IC(50) values of 3.2, 1.4, 8.9, 0.2, and 0.3 microM were obtained for the standard isozyme inhibitors CYP1A2/furafylline, CYP2C9/sulfaphenazole, CYP2C19/tranylcypromine, CYP2D6/quinidine, and CYP3A4/ketoconazole, respectively, which were in good agreement with literature values. trans-Resveratrol showed IC(50) values of 11.6 microM for CYP2C19 and 1.1 microM for CYP3A4, but the IC(50) values exceeded 50 microM for all the other CYP isozymes, which indicated no inhibition. No enzyme inhibition was observed for trans-resveratrol-3-sulfate. Our results indicate that trans-resveratrol is a marginal inhibitor of CYP3A4 and a weak inhibitor of CYP2C19, but its major metabolite trans-resveratrol-3-sulfate is not an inhibitor of any of the cytochrome P450 isozymes investigated.