普兰林肽的固相合成  

Solid phase synthesis of pramlintide

作　　者：韩月[1] 尹志峰[1] 赵红玲[1] 王小青[1] 王良友[1] 

机构地区：[1]承德医学院中药研究所,承德067000

出　　处：《中国新药杂志》2012年第9期1046-1049,共4页Chinese Journal of New Drugs

摘　　要：目的:探索普兰林肽的固相合成、氧化条件及纯化方法。方法:采用Fmoc固相多肽合成法,以Rink Amide-AM树脂做载体,HBTU/HOBt/DIEA做缩合剂,逐步缩合得到全保护线性普兰林肽树脂,以TFA/苯甲硫醚/苯酚/H2O/EDT/TIS配比的裂解液脱除保护基团,分别采用空气,二甲基亚砜,双氧水氧化两个半胱氨酸的巯基形成一对二硫键,半制备反相高效液相色谱法纯化。结果:合成含37个氨基酸以及一对二硫键的普兰林肽经RP-HPLC和MALDI-TOF-MS确证,粗品纯度在50.0%以上,粗品经半制备型反相高效液相色谱纯化,所得精肽的纯度大于95.0%,总收率为30.5%。结论:该方法简单,合成的产品成本低,纯度高,可为工业化生产提供借鉴。Objective：To develop the solid phase synthesis method of pramlintide and to establish the oxidation and purification condition.Methods：Pramlintide was synthesized by Fmoc solid phase peptide synthesis method,using Rink Amide-AM Resin as solid carrier and HBTU/HOBt/DIEA as coupling reagents.Protection groups of the crude peptide were cleaved by a mixture of TFA/Thioanisole/phenol/H2O/EDT/TIS.Disulfide bridge was formed by air,DMSO or H2O2 oxidation.The crude pramlintide was purified by semi-preparative RP-HPLC.Results：Pramlintide was identified by RP-HPLC and MALDI-TOF-MS.The purity of the crude peptide was about 50.0%.After the purification by semi-preparative HPLC,the purity was increased to more than 95.0%,and the overall yield was 30.5%.Conclusion：This method is simple and cost-effective and can produce pramlintide with high purity,which may be useful for the industrial manufacturing.

关 键 词：普兰林肽 固相多肽合成 二硫键形成 

分 类 号：R914.5[医药卫生—药物化学]