蛇床子素对基因多态性的CYP2C9酶活性的作用及对吲哚美辛药代动力学的影响研究

发布时间：2018-02-15 09:36

本文关键词： CYP2C9 蛇床子素基因多态性　出处：《重庆医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：蛇床子素是存在于多种中药中的药理活性成分，具有广泛的药理作用，在临床应用前景广阔，是一个极具开发利用价值的药理活性成分。目前，国内外对蛇床子素药理药效方面的研究较多，而关于蛇床子素药物代谢动力学相关研究较少，本文了针对蛇床子素代谢的过程，考察其对药物代谢酶CYP2C9的作用，为蛇床子素的开发应用提供参考。 CYP2C9是在人群中存在基因多态性，常见基因型为CYP2C9*1、CYP2C9*2、CYP2C9*3。与CYP2C9*1相比，突变型的CYP2C9*2、CYP2C9*3酶代谢能力降低，，但对药物的反应灵敏性增加，更易受到药物的诱导或抑制，进一步使酶代谢药物的能力降低，从而导致药物不良反应的发生。因此，考察蛇床子素对不同基因型的CYP2C9酶的作用，对不同患者间的安全用药具有重要意义。本文从体外初步评价了蛇床子素对CYP2C9*1、CYP2C9*2、CYP2C9*3酶的作用及作用机制，发现蛇床子素抑制CYP2C9*1、CYP2C9*2、CYP2C9*3酶活性，且抑制作用表现为：CYP2C9*3CYP2C9*2CYP2C9*1，抑制参数分别为Ki=21.93μM（CYP2C9*1）、Ki=18.10μM（CYP2C9*2）、Ki=13.12μM（CYP2C9*3）。表明蛇床子素对CYP2C9酶的抑制作用存在个体差异。 CYP2C9在不同种属间的表达或酶活性高低存在差异，为了明确蛇床子素在混合微粒体中对CYP2C9酶的作用，本文同时考察了蛇床子对大鼠和人的混合肝微粒体中CYP2C9酶的作用。发现蛇床子素对两者中CYP2C9酶均具有抑制作用，且种属差异明显：对大鼠肝中CYP2C9酶呈竞争性抑制，而在人肝中呈非竞争性抑制；且在大鼠肝体系中的抑制作用（Ki=8.187μM）显著大于人肝体系中的抑制作用（Ki=49.81μM）。药物在体内需经复杂的转化过程，因此，药物对酶的作用不能简单地从体外实验结果断定，需进一步进行体内验证。本文选用主要经CYP2C9代谢的吲哚美辛，与蛇床子素联合给药于大鼠后，给药组与对照组药动学参数比较发现：吲哚美辛的AUC、Cmax、CL/F不变，Tmax、T1/2延长，其表明蛇床子素对吲哚美辛的体内药动学过程存在影响，但对CYP2C9介导的代谢过程无作用。由于体外和体内实验的差异性，其蛇床子素对吲哚美辛体内其他过程的作用需进一步研究明确。本文通过体外、体内研究了蛇床子素对CYP2C9酶的抑制作用，并发现个体差异和种属差异的影响。结果提示在蛇床子素实验研究和临床使用中，需注意个体差异和种属差异产生的影响。蛇床子素影响吲哚美辛的体内药动过程，因而在临床用药时，尤其注意基因型为CYP2C9*2、CYP2C9*3的患者使用药物的风险，减少药物不良反应，并避免与其他药物发生相互作用，保证临床安全合理用药。
[Abstract]:Osthol is a kind of pharmacological active component which exists in many kinds of traditional Chinese medicine. It has extensive pharmacological effect and has a broad prospect in clinical application. It is a pharmacological active component with great value of development and utilization. There are many studies on the pharmacological effects of osthol at home and abroad, but there are few studies on the pharmacokinetics of osthol. In this paper, the effects of osthol on the metabolism of osthol CYP2C9 were investigated. It provides a reference for the development and application of osthol. The common genotype of CYP2C9 is CYP2C9C9O2CYP2C9O2CYP2C9O33.Compared with CYP2C9*1, the metabolic ability of mutant CYP2C9O2CYP2C9t3 is decreased, but its sensitivity to drugs is increased, and it is more susceptible to drug induction or inhibition. The ability of metabolizing drugs was further reduced, which led to the occurrence of adverse drug reactions. Therefore, the effects of osthol on CYP2C9 enzymes of different genotypes were investigated. In this paper, the effect and mechanism of osthol on CYP2C9O2-CYP2C9O3-CYP2C9O3-CYP2C91-CYP2C91-CYP2C91-CYP2C93-CYP2C93-was evaluated in vitro, and it was found that osthol inhibited the activity of CYP2C9O2-CYP2C93. The inhibitory effects were as follows: 1: CYP2C9, 3CYP2C9, 2CYP2C9, and the inhibitory parameters were Ki=21.93 渭 M9, CYP2C9, CYP2C9 / 1, respectively. The inhibitory effect of osthol on CYP2C9 enzyme was different in individuals, and the inhibitory effect of CYP2C9 / 2 was 13.12 渭 MCYP2C9 / 1, which indicated that the inhibitory effect of osthol on CYP2C9 was different in individuals. In order to clarify the effect of osthol on CYP2C9 enzyme in mixed microsomes, the expression or enzyme activity of CYP2C9 was different among different species. The effects of osthol on CYP2C9 enzyme in rat and human liver microsomes were also investigated. It was found that osthol could inhibit CYP2C9 enzyme in both of them, and the difference of species was significant: the inhibition of CYP2C9 enzyme in rat liver was competitive. The inhibitory effect in rat liver system was 8.187 渭 M), and the inhibitory effect in human liver system was significantly higher than that in human liver system (49.81 渭 M-1). The drug needs complex transformation process in vivo. Therefore, the effect of drugs on enzyme can not be simply determined from the results of in vitro experiments, and should be further verified in vivo. In this paper, indomethacin, which is mainly metabolized by CYP2C9, is selected. The pharmacokinetic parameters of indomethacin group and control group were compared with those of control group. The results showed that the CLF of indolomethacin was prolonged, which indicated that osthol had an effect on the pharmacokinetic process of indomethacin in vivo, and the effect of osthol on the pharmacokinetic process of indomethacin in vivo was observed. Because of the difference between in vitro and in vivo experiments, the effect of osthol on other processes of indomethacin in vivo should be further studied. The inhibitory effect of osthol on CYP2C9 enzyme was studied in vitro, and the effects of individual and species differences were found. Attention should be paid to the effects of individual and species differences. Osthol affects the pharmacokinetics of indomethacin in vivo. Therefore, in clinical use, particular attention is paid to the risk of drug use in patients with genotypes CYP2C9 and CYP2C9, so as to reduce adverse drug reactions. And avoid the interaction with other drugs to ensure clinical safety and rational use.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R969.1

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