Eptifibatide的固相合成及分离纯化  

Fmoc Solid-phase Synthesis and Purification of Eptifibatide

作　　者：熊瑛[1] 王宇[1] 陈亮[1] 

机构地区：[1]厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2007年第1期100-103,共4页Journal of Xiamen University：Natural Science

摘　　要：Eptifibatide是一种血小板糖蛋白GPⅡb/Ⅲa受体拮抗剂,临床研究结果显示其医用上的产业前景良好.采用Rink-Am-Resin树脂作为固相载体,Fmoc为保护策略固相合成了Eptifibatide.以DMF为反应溶剂,Piperidine-DMF脱去Fmoc保护基团,以TBTU/HOBt/DIEA作为肽键缩合试剂进行接肽反应,用TFA/EDT/TIS/H2O定量地从树脂上切除,制备型高效液相色谱分离纯化粗肽.同时对产品进行色谱分析和质谱鉴定.实验结果表明Fmoc合成策略中各步的缩合率均在95%以上,多数在99%以上.粗肽纯度为86.8%,产率为55%.分离纯化后的产物纯度达到98.5%,MOLDI-TOF质谱测得的m/z与理论值相符.为化学合成Eptifibatide及其分离纯化提供了一条切实可行的路线.Eptifibatide is the receptor antagonist of platelet glycoprotein GP Ⅱ b/Ⅲ a,Clinical study showed that it has the advantage of potent effect. In the present work,a strategy for the synthesis and purification of Eptifibatide was presented. Eptifibatide has been synthesized by solid - phase method using the base-labile Fmoc group for protecting the a-amino acid. The peptide was assembled on Rink-Am-Resin. DMF is used as the coupling solvent; Piperidine-DMF to removed the Fmoc group;TBTU/HOBt/DIEA as the activation system,and TFA/EDT/TIS/H2O as the cleavage condition. The synthetic product was purified by prepared reversedphase high performance liquid chromatography and identified on MALDI-TOF-MS and HPLC. The experimental result shows that the ratio of the coupling synthesis of each amino acid is over 95 % ,most of them are over 99 %. After cleavage of peptide resin,the purity of rude Eptifibatide was about 86. 8%, and the yield of it is 55 %. The final purity of our product was found more than 98. 5%. The molecular mass of the product is correspond to molecular mass of the Eptifibatide. This synthetic strategy is practicable for chemical synthesis and purification of Eptifibatide.

关 键 词：EPTIFIBATIDE 固相合成 Fmoc法 

分 类 号：Q958.1[生物学—动物学]