论文总字数：17469字

摘 要

索非布韦（sofosbuvir，SOF）是世界上第一个无需结合使用干扰素即可实现全口服治疗丙肝的药物，是丙型肝炎病毒（HCV）特异性NS5B聚合酶的核苷抑制剂。临床实验表明，SOF和利巴韦林二者联合全口服方案可用于治疗基因Ⅱ型、Ⅲ型丙肝病毒的感染，用药十二周总体的SVR为50％～78％。SOF结合PEG-IFN及Ribavirin可用于基因Ⅰ型及Ⅳ型HCV的感染，用药十二周的SVR达90％。SOF由于其服用方便，治愈率高，不良反应发生率低，使其极有可能成为“重磅炸弹”药物。

本文建立一HPLC法用于检测样品中索非布韦中间体SF-X1的含量。本文采用Aglient ZORBAX XDB-C18色谱柱，以H₂O为流动相A相，ACN/H₂O（9：1）为流动相B相，色谱柱两端温度为30摄氏度，每分钟流速为1mL ，检测波长230 nm，在这一色谱条件下，索非布韦中间体与杂质具有良好的分离度；索非布韦中间体在 0.05~0.25 mg/mL内 , 线性关系良好 ,r = 0.9999 ,平均回收率为99.6 % ,RSD 为 0.24 %(n = 6) ；此法实验步骤简单、数据准确，灵敏度高，可用来测定874638-80-9的含量。

关键词 : HPLC; 索非布韦 ; 索非布韦中间体 ; 方法建立；方法验证

HPLC method for the determination of sofosbuvir intermediates content in the samples

Abstract

Sofosbuvir(SOF)，a novel hepatitis C virus (HCV) nucleoside analog NS5B polymerase inhibitor，might be the first drug on the globe that achieved an INF free all-oral regimen in the treatment of chronic hepatitis C (CHC) patients with genotypes 2 and 3 infections，Twelve weeks after medication,overall sustained virological response (SVR) was 50％～78％, SOF combined with peginterferon (PEG-IFN) and ribavirin could be used for the treatment of CHC patients with genotypes 1 and 4 infections ,and 12 weeks after medication, SVR was as high as 90％. Having the features of easy administration ,high cure rate and low rate of adverse reactions, SOF Will be likely to be the most potent “blockbuster” type drug for the treatment of disease.

In this paper, to establish a HPLC method for determination of content of Sofosbuvir intermediates(SF-X1) in the samples.This method uses the chromatographic column filled with octadecyl bonded silica gel, and with water as mobile phase A, as acetonitrile/water (9:1) as mobile phase B, the column temperature was 30, the flow rate was 1 mL/min, the detection wavelength of 230 nm. In this chromatographic condition, Sofosbuvir intermediates and impurities were well separated; In the scope of 0.05 ~ 0.25 mg/mL, Sofosbuvir intermediates had a good linear relation, r = 0.9999, the average recovery was 99.81%, RSD was 0.24% (n = 6); This procedureof the method is simple and the data is accurate,the system has high sensitivity,so it can be used as to determine the content of 874638-80-9.

Keywords: HPLC; Sofosbuvir; Sofosbuvir intermediates; Method establishment; Method validation

目录

摘 要 I

Abstract II

第一章 引 言 1

1.1选题背景 1

1.2选题意义 1

第二章 新型抗丙型肝炎病毒药索非布韦 3

2.1 索非布韦 3

2.1.1 索非布韦简介 3

2.1.2 作用机制 4

2.1.3剂量和给药方法 4

2.1.4药代动力学 4

2.1.5癌变、基因突变、遗传毒性 4

2.1.6动物毒理学 5

2.1.7 耐药性 5

2.2 索非布韦中间体简介 5

第三章 HPLC法测定样品中索非布韦中间体含量的方法学研究 6

3.1引言 6

3.2 高效液相色谱法的发展与构建 6

3.3实验准备 7

3.3.1实验仪器 7

3.3.3实验试剂 7

3.4实验过程 7

3.4.1 色谱条件 7

3.4.2 方法的专属性考察 8

3.4.3 对照品溶液的制备 9

3.4.4 供试品溶液的制备 9

3.4.5 检出限与定量限 9

3.4.6线性关系考察 11

3.4.7 进样精密度实验 11

3.4.8 重复性实验 11

3.4.9 稳定性试验 12

3.4.10 回收率实验 12

3.5实验结果与讨论 13

3.5.1紫外-可见光谱（UV-Vis 谱）及测定波长的选择 13

3.5.2 流动相及比例的选择 13

3.5.3流速的选择 13

3.5.4进样量的选择 14

3.5.5 洗脱方式的选择 14

3.5.6 外标法测定含量 15

3.6 874638-80-9含量方法学验证结果 15

第四章 结论与展望 16

参考文献 17

致 谢 18

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