瑞莫瑞林的Fmoc固相合成及其活性检测

Fmoc solid phase synthesis and activity determination of relamorelin

目的 优化瑞莫瑞林固相合成策略,并对其进行活性检测。方法 以Fmoc为氨基酸保护基团,采用HOBt/HBTU/DIEA复合型缩合体系,以逐个延伸Fmoc固相合成法合成瑞莫瑞林,并优化裂解条件,通过GHSR1a体外激动活性评价实验验证瑞莫瑞林的生物学活性。结果 最优裂解液条件为TFA∶TIS∶H_(2)O=95∶2.5∶2.5,合成瑞莫瑞林收率为77.5%。合成的粗肽经高效制备色谱纯化及反相高效液相色谱分析后,纯度为99.3%(220 nm)。目标产物结构经LC-MS、HR-ESI-MS、1H-NMR分析确证。合成的瑞莫瑞林对GHSR-1a激动活性的EC_(50)值为1.298 nmol·L^(-1),与文献报道活性相当。结论 瑞莫瑞林的Fmoc固相合成方法具有快捷、高效、反应条件温和等特点,适合大批量生产瑞莫瑞林。

This study aims to optimize the solid phase synthesis strategy of relamorelin and evaluate its biological activity.The solid-phase synthesis was performed using Fmoc as the amino acid protecting group and HOBt/HBTU/DIEA as the complex condensation system.The lysate was chosen as the mixture of TFA:TIS:H_(2)O=95∶2.5∶2.5.The obtained crude peptide was purified by high performance preparative chromatography to afford the final product with a purity of 99.3%(220 nm) and yield of 77.5%.We further assayed the agonist activity of this peptide with the GHSR-1 a activation(EC_(50)=1.298 nmol·L^(-1)),which was comparable to that reported in the literature.This synthetic method is rapid, efficient, and requires mild reaction conditions, which is suitable for large-scale production of relamorelin.