Cocktail探针底物法研究白花丹醌对CYP和UGT酶的作用机制

发布时间：2018-01-12 10:19

  本文关键词：Cocktail探针底物法研究白花丹醌对CYP和UGT酶的作用机制　出处：《华东师范大学》2017年硕士论文　论文类型：学位论文

  更多相关文章： cocktail方法 白花丹醌 CYP酶 UGT酶 药物-药物相互作用

【摘要】：随着中药和西药联用的普及,它们之间发生相互作用的报道越来越多,而CYP和UGT酶介导了大部分的中药-西药相互作用。本研究在体外建立了 cocktail探针底物法,用于考察化合物对CYP和UGT酶的活性影响,评估潜在的药物-药物相互作用,为新药开发和临床安全用药提供指导。首先,本研究在人和大鼠肝微粒体中分别建立了研究CYP和UGT酶活性的cocktail方法,其中CYP酶主要包括 CYP1A2、CYP2B6/1、CYP2C9/11、CYP2D6/1、CYP2E1 和 CYP3A4/2;UGT 酶主要包括 UGT1A1、UGT1A3、UGT1A6、UGT1A9/PROG 和UGT2B7/AZTG。然后利用此方法评价了多种中药单体对CYP和UGT酶活性的影响,发现白花丹醌对CYP酶活性有较强的抑制作用,并探讨了作用机制。白花丹醌是从白雪花中提取的具有抗炎和抗过敏等药理活性的中药单体,近年来还发现有较强的抗肿瘤作用。然而白花丹醌在药物代谢方面的研究甚少,尤其对Ⅰ相CYP酶和Ⅱ相UGT酶的活性影响及作用机制未见报道。本论文发现白花丹醌对六种CYP酶均有较强的抑制效果,并且为非时间依赖的可逆抑制。酶动力学研究显示,在人肝微粒体中,白花丹醌对CYP2B6、CYP2C9、CYP2D6、CYP2E1和CYP3A4呈现混合型抑制,而对CYP1A2呈现非竞争性抑制,Ki值分别为1.62μM、2.16μM、1.46μM、0.65μM、0.88μM和 0.15μM;在大鼠肝微粒体中,白花丹醌对CYP1A2和CYP2D1呈现混合型抑制,对CYP2B1、CYP2C11和CYP2E1呈现竞争性抑制,Ki 值分别为 2.64 μM、9.49 μM、9.93 μM、7.85 μM 和 6.28 μM,而对CYP3A2表现出非典型酶动力学抑制。此外,白花丹醌对Ⅱ相UGT酶无明显抑制效果。综上所述,本研究在人和大鼠肝微粒体中成功构建了检测CYP和UGT酶活性的cocktail探针底物法,并开展了对多种中药单体的评价和研究。本论文证明了白花丹醌对人和大鼠肝微粒体中六种CYP酶均有较强的抑制效果,而对UGT酶活性无明显影响,为进一步开发及合理使用白花丹醌提供了重要信息。
[Abstract]:With the popularization of traditional Chinese medicine and western medicine, there are more and more reports of interaction between them. However, CYP and UGT enzyme mediated the interaction between traditional Chinese medicine and western medicine. In this study, cocktail probe substrate method was established in vitro. It can be used to investigate the effects of compounds on the activity of CYP and UGT enzymes, to evaluate the potential drug-drug interactions, and to provide guidance for the development of new drugs and clinical safe use of drugs. In this study, a cocktail method was established to study the activities of CYP and UGT in human and rat liver microsomes, in which CYP enzymes mainly included CYP1A2CYP2B6 / 1. CYP2C9 / 11, CYP2D6 / 1, CYP2E1 and CYP3A4 / 2; UGT enzymes mainly include UGT1A1, UGT1A3, UGT1A6. UGT1A9/PROG and UGT2B7 / AZTG.Then the effects of various traditional Chinese medicine monomers on the activities of CYP and UGT were evaluated by this method. It was found that Dandanquinone had a strong inhibitory effect on the activity of CYP and discussed its mechanism. It was a traditional Chinese medicine monomer with anti-inflammatory and anti-allergic activities extracted from Flos Lonicerae. In recent years, it has been found that it has strong anti-tumor effect. However, there are few studies on the drug metabolism of Danquin. In particular, there was no report on the effect and mechanism of the activity of phase 鈪，

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