Metabolism of the phytoestrogens by sulfate and glucuronic acid conjugation reactions

通过硫酸盐和葡萄糖醛酸缀合反应进行植物雌激素的代谢

• 批准号: 14572072
• 负责人: OGURA Kenichiro
• 金额: $ 1.92万
• 依托单位: Tokyo University of Pharmacy and Life Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572072/
• 关键词: phytoestrogen; genistein; daidzein; sulfate conjugation; glucuronic acid conjugation; metabolism; excretion; conjugation reaction

Regioselective glucuronidation and formation of sulfoglucuronides of the phytoestrogen genistein (GS,5,7,4'-trihydroxyisoflavone) was investigated using human liver cytosol, microsomes, purified recombinant human sulfotransferase(SULT) isoforms, SULT1A1,SULT1A3,SULT2A1, and SULT1E1, and recombinant human glucuronosyltransferases (UGT) isoforms, UGT1A1,UGT1A3,UGT1A4,UGT1A6,UGT1A9,UGT2B7 and UGT2B15. 7-Position-preferential glucuronidation of GS was observed in human hepatic microsomes from 7 female and 3 male subjects. Average ratios for 7- to 4'-glucuronide formation were 26:1 from GS in these human liver microsomes. Apparent Km value for the 7- and 4'-glucuronidation of GS by the microsomes from a male subject was 12.3 and 84.2 μM, respectively. Recombinant human UGTs, UGT1A1,UGT1A3,UGT1A6,UGT1A9, and UGT2B7 had activity for 7-glucuronidation of GS. However, no isoform showed catalytic activity of 4'-glucuronidation of GS. UGT1A1 and UGT1A9 exhibited much higher catalytic efficiency, kcat/Km, for 7-glucuronidation than did the other isoforms. UGT1A9 showed a Km value of 9.5 for the mono-glucuronidation of GS, which were very similar to that of human microsomes. The observed kcat/Km indicated that UGT1A1 and UGT1A9 catalyzed 7-glucuronidation of GS in the human liver microsomes. The results of enzymatic study using synthetic GS 7- and 4'-glucuronides as substrates for sulfation by recombinant SULTs showed that SULT1E1 most efficiently catalyzed the formation of sulfoglucuronides from GS mono-glucuronides. On the other hand, only UGT1A1 showed a catalytic activity of the sulfoglucuronide formation from GS 4'-sulfate. No isoform had activity to form sulfoglucuronide from GS 7-sulfate.

使用人肝细胞质、微粒体、纯化的重组人磺基转移酶 (SULT) 亚型、SULT1A1、SULT1A3、SULT2A1 和 SULT1E1，研究了植物雌激素金雀异黄酮（GS,5,7,4'-三羟基异黄酮）的区域选择性葡萄糖醛酸化和磺基葡萄糖苷酸的形成，以及重组人葡萄糖醛酸基转移酶 (UGT) 亚型、UGT1A1、UGT1A3、UGT1A4、UGT1A6、UGT1A9、UGT2B7 和 UGT2B15 在 7 名女性和 3 名男性受试者的人肝微粒体中观察到 7 至 4' 的平均比率。 -葡萄糖苷酸这些人肝微粒体中 GS 的形成比例为 26:1，来自男性受试者的微粒体对 GS 进行 7-和 4'-葡萄糖醛酸化的表观 Km 值分别为 12.3 和 84.2 μM。 UGT1A6、UGT1A9 和 UGT2B7 具有以下活性： GS 的 7-葡萄糖醛酸化作用然而，没有同种型显示 GS 的 4'-葡萄糖醛酸化催化活性，UGT1A9 对 7-葡萄糖醛酸化显示出比其他同种型高得多的 Km 值。 GS 的单葡萄糖醛酸化为 9.5，非常相似观察到的 kcat/Km 表明 UGT1A1 和 UGT1A9 在人肝微粒体中催化 GS 7-葡萄糖醛酸化。使用合成 GS 7-和 4'-葡萄糖醛酸作为重组 SULT 硫酸化的底物的酶学研究结果。结果表明 SULT1E1 最有效地催化磺基葡萄糖醛酸苷的形成另一方面，只有UGT1A1显示出从GS 4'-硫酸盐形成磺基葡萄糖苷酸的催化活性，没有异构体具有从GS 7-硫酸盐形成磺基葡萄糖醛酸苷的活性。

项目成果

H.Nakano, et al.: "Regioselective monosulfation and disulfation of the phytoestrogens daidzein and genistein by human liver sulfotransferases"Drug Metabol.Pharmacokin.. 印刷中. (2004)

H.Nakano 等人：“人肝磺基转移酶对植物雌激素黄豆苷元和金雀异黄素的区域选择性单硫酸化和二硫酸化”Drug Metabol.Pharmacokin.. 出版中（2004 年）。

Takahito Nishiyama: "Sulfation of Environmental Estrogens by Cytosolic Human Sulfotransferases"Drug Metabolism and Pharmacokinetics. 17(3). 221-228 (2002)

Takahito Nishiyama：“胞质人磺基转移酶对环境雌激素的硫酸化”药物代谢和药代动力学。