维兰特罗关键中间体及有关物质的高效液相色谱分析方法优化

Optimization of HPLC Method for the Analysis of Key Intermediate of Vilanterol and Its Related Substances

摘要: 维兰特罗是一种治疗慢性阻塞性肺病(COPD)的β₂受体激动剂，其起效作用快、持续时间长、耐受性好而备受关注。2-(2,2-二甲基-4H-1,3-苯并二氧杂环己烯-6-基)-2-羰基乙基氨基甲酸叔丁酯是合成维兰特罗的关键中间体之一，德信佳生物医药公司开发了一种绿色合成工艺，该路线经由五步反应，共涉及5个中间产物，即中间体1、2、3、4、5。本研究拟建立关键中间体5 (2-(2,2-二甲基-4H-1,3-苯并二氧杂环己烯-6-基)-2-羰基乙基氨基甲酸酯)及其有关物质(或杂质)的HPLC分析方法，通过流动相组成及比例、检测波长等条件筛选与优化，中间体1、2、4、5主成分及有关物质可以实现有效分离；利用上述建立的HPLC方法，对两批次中间体5产品的有关物质进行分析，并采用峰面积归一化法计算含量。它们均检测到三个有关物质，其总和低于1.5%，该研究为后续关键中间体及原料药中有关物质的制备分离、结构推测及定量分析提供了方法参考。

Abstract: Vilanterol, a β₂ receptor agonist inhibiting chronic obstructive pulmonary disease (COPD), has been respected for rapid effect, long duration and good tolerance. 2-(2,2-dimethyl-4H-1,3-benzodioxane heterocyclohexene-6-yl)-2-carbonyl ethyl carbamate is one of the key intermediates of vilanterol, which has developed by Dexinjia Biopharm Co., Ltd. The green preparation route is divided into five-step reactions involving five intermediates, namely intermediate 1, 2, 3, 4, 5. In this study, the HPLC analysis method of intermediate 5 (2-(2,2-dimethyl-4H-1,3-benzodioxane heterocyclohex-ene-6-yl)-2-carbonyl ethyl carbamate) and its related substances (or impurities) was optimized mainly through the composition and proportion of mobile phase and detection wavelength. As a result, the intermediate 1, 2, 4, 5 and related substances can be effectively separated. Using the es-tablished HPLC method, the related substances of two batches of intermediate 5 were analyzed, three major related substances were detected and the total content was less than 1.5% according to the peak area normalization. These investigations provided experimental methods for the further preparation, structural speculation and quantitative analysis of related substances in the key in-termediates and raw drugs of vilanterol.