氯唑沙宗在SD大鼠体内的代谢及药动学相互作用研究

发布时间：2018-06-18 07:58

本文选题：氯唑沙宗 + HPLC-PDA-ESI-ITMS~n　；参考：《福建中医药大学》2014年硕士论文

【摘要】：氯唑沙宗(chlorzoxazone)的化学名为5-氯-2-苯并恶唑酮(5-chloro-2-Hydroxy-benzoxazole),是一种口服中枢性肌肉松弛剂,作用快且持久,可有效缓解各种急慢性扭伤、挫伤、肌肉劳损等引起的软组织疼痛以及中枢神经引起的肌肉痉挛疼痛。氯唑沙宗作用于脊髓反射中枢,能够阻断连接知觉神经和运动神经的介在神经元。由于多突触反射的抑制,使反射兴奋低下而缓解骨骼肌紧张。另外氯唑沙宗常常用于评价肝脏微粒体细胞色素CYP2EI酶活性,作为其唯一的特异性探针底物,氯唑沙宗在人体内主要经羟基化代谢为6-羟基氯唑沙宗,根据6-羟基氯唑沙宗与氯唑沙宗的血药浓度比值或尿中6-羟基氯唑沙宗的回收率可反应CYP2EI的酶活性。实验第一部分通过采用HPLC-PDA-ESI-ITMS"联用技术,分离并鉴定出SD大鼠口服氯唑沙宗原料药后,尿液中的5种氯唑沙宗代谢产物,分别为6-羟基氯唑沙宗、6-羟基氯唑沙宗葡萄糖醛酸轭合物、6-羟基氯唑沙宗硫酸酯轭合物、氯唑沙宗葡萄糖醛酸轭合物和氯唑沙宗硫酸酯轭合物,同时揭示了这5种代谢产物的质谱裂解规律。实验发现6-羟基氯唑沙宗葡萄糖醛酸轭合物和6-羟基氯唑沙宗硫酸酯轭合物为主要代谢产物,可以经过水解酶的作用得到6-羟基氯唑沙宗,所以实验第二部分设计了一套方案,获取6-羟基氯唑沙宗葡萄糖醛酸轭合物和6-羟基氯唑沙宗硫酸酯轭合物这两个目标代谢产物。实验第二部分我们通过给SD大鼠灌胃一定剂量的氯唑沙宗原料药后,收集其尿液。采用上样过D-101大孔树脂方法去除尿液中的部分杂质,得到一系列洗脱流份。与此同时结合高效液相色谱仪的高效分离检测手段,检测样品,合并含有相同代谢产物的流份,得到一定纯度的代谢产物粗提物。然后采用半制备型高效液相色谱仪对已经得到的代谢产物粗提物进行分离制备,从而得到所需的目标代谢产物。最后对所制备得到的目标代谢产物进行含量纯度测定检验和质谱结构确证。第三部分,我们考察了临床上常用复方氯唑沙宗片中的两主成分氯唑沙宗和对乙酰氨基酚在SD大鼠体内联合用药的药动学行为进行了初步探讨,为临床合理用药提供了指导。实验结果发现与单独用药相比,联合用药后,对乙酰氨基酚的达峰浓度显著降低、平均滞留时间有所增加。氯唑沙宗的半衰期有所增加。
[Abstract]:Chlorzoxazone (Chlorzoxazone) is an oral central muscle relaxant called 5-chloro-2-Hydroxy-benzoxazoleone. It has a fast and lasting effect on relieving all kinds of acute and chronic sprain and contusion. Soft tissue pain caused by muscle strain and muscle spasm caused by central nervous system. Clozoxazone acts on the spinal reflex center and blocks the mediators connecting the perceptual and motor nerves. Because of the inhibition of polysynaptic reflex, the reflex is low and the skeletal muscle tension is alleviated. In addition, chlorzoxazone is often used to evaluate the activity of liver microsomal cytochrome CYP2EI. As its only specific probe substrate, clozoxazone is mainly metabolized by hydroxylation to 6-hydroxychlorazolazone in human body. The activity of CYP2EI can be reflected by the ratio of 6-hydroxyclozoxazone to clozoxazone in blood or the recovery rate of 6-hydroxyclozoxazone in urine. The first part of the experiment used HPLC-PDA-ESI-ITMS to isolate and identify five chlorzoxazone metabolites in urine after oral clozoxazone in SD rats. They are 6-hydroxyclozoxazole-6-hydroxychloroxazole-glucuronic acid yoke, chlorzoxazone glucuronic acid conjugate and chlorzoxazolone sulfate yoke, respectively. At the same time, the law of mass spectrometry cleavage of the five metabolites was revealed. It was found that 6-hydroxyclozoxazone glucuronic acid conjugate and 6-hydroxyclozoxazone sulfate conjugate were the main metabolites, and 6-hydroxychlorazolazone could be obtained by hydrolase. Therefore, a set of schemes was designed in the second part of the experiment. Two target metabolites of 6-hydroxyclozoxazone glucuronic acid conjugate and 6-hydroxyclozoxazone sulfate conjugate were obtained. In the second part of the experiment, we collected the urine of SD rats by intragastric administration of clozoxazone. A series of elution fractions were obtained by using D-101 macroporous resin to remove some impurities in urine. At the same time, combined with the high performance liquid chromatograph, the sample was detected, combined with the same metabolites of the flow, a certain purity of the crude extract of metabolites was obtained. Then the crude extracts of metabolites were separated and prepared by semi-preparation high performance liquid chromatograph (HPLC), and the desired metabolites were obtained. Finally, the content purity of the target metabolites was determined and the structure of the target metabolites was confirmed by mass spectrometry. In the third part, we investigated the pharmacokinetic behavior of acetaminophen combined with chlorzoxazone Zong He, which is commonly used in clinical compound chlorzoxazone tablets in SD rats, and provided guidance for rational use of acetaminophen in clinic. The results showed that the peak concentration of acetaminophen was significantly decreased and the mean residence time was increased. The half-life of chlorzoxazone increased.
【学位授予单位】：福建中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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