多黏菌素E类似物的纯化和结构鉴定-中国抗生素杂志.PDF

中国抗生素杂志2018年1月第43卷第1期 . 59 . 文章编号：1001-8689(2018)01-0059-05 分离纯化与化学合成 多黏菌素E类似物的纯化和结构鉴定 吴勇 周梦漪 薛春佳 冯军* ( 中国医药工业研究总院上海多米瑞生物技术有限公司，上海201203) 摘要：目的 对硫酸多黏菌素中的低含量成分进行纯化和鉴定。方法 利用两次反相色谱纯化制备得到单组份，并通过 Nα-(2,4-二硝基-5-氟苯基)-L-丙氨酰胺(Nα-(2,4-Dinitro-5-fluorophenyl)-L-alaninamide, FDAA)试剂衍生法研究组分的组成氨基酸及 其构型。基于二级质谱和氨基酸组成的结果对化合物进行了结构鉴定。结果 多黏菌素E2 的氨基酸组成、分子量与文献报道一 致；杂质3和杂质9结构中含有L-Leu残基和羟基化的脂肪酸链；杂质6含有L-Val残基。结论 杂质3 、杂质6和杂质9分别为3-OH- E2 、Val-E2和3-OH-E 1。 关键词：多黏菌素E ；纯化；结构鉴定 中图分类号：R978.1 文献标志码：A Purification and structure elucidation of colistin E analogues Wu Yong, Zhou Meng-yi, Xue Chun-jia and Feng Jun (Shanghai Domiry Bio-tech Co. ,Ltd, China State Institute of Pharmaceutical Industry, Shanghai 201203) Abstract Objective The purification and identification of minor components in colistin E. Methods Each single component was purified by using reverse-phase chromatography twice, and derivatization using the reagent FDAA was performed to analyze the amino acid composition and the stereochemistry of each component. Structural identification was based on the secondary mass spectrometry and amino acid composition. Results The amino acid composition and molecular weight of colistin E2 were consistent with those published reports. Both impurities 3 and 9 contained L-Leu residues and hydroxylated fatty acid chains. The impurity 6 contained an L-Val residue. Conclusion Impurity 3, impurity 6, and impurity 9 were 3-OH-E2, Val-E2, and 3-OH-E1, respectively. Key words Colistin E; Purification; Structure elucidation 多黏菌素E(colistin E)是一类由多黏芽孢杆菌黏菌 构见图1。临床使用的多黏菌素E有两种形式，一种 素亚种(Bacillus polymyxa subsp . colistinus)代谢产生的 是口服或局部使用的硫酸多黏菌素E ；另一种