摘要
目的探索利那洛肽的固相合成工艺及环化方法,制备得到高纯度的利那洛肽。方法采用Fmoc固相多肽合成法,以Wang树脂为载体,HOBt/DIC为缩合剂,逐步缩合得到利那洛肽线性肽树脂。在此基础上,采用固相-裂解-液相结合的方式分两次完成三对二硫键的配对。粗品经反相高效液相色谱法纯化后得到目标产物。结果利那洛肽粗品纯度为75.4%,经反相高效液相色谱法纯化后,精肽纯度高达99.32%。产物经高分辨质谱表征,与标准品色谱特征一致。结论利用该方法,能够简单高效地实现利那洛肽的大规模制备,为工业化生产提供借鉴。
OBJECTIVE To explore the solid-phase technics and cyclization method of linaclotide for obtaining high purity of linaclotide.METHODS The linear peptide resin of linaclotide was assembled by Fmoc solid-phase peptide synthesis method using Wang resin as solid phase and HOBt/DIC as condensing agents.Three disulfide bonds were built in two steps by combinated solid phase method-cleavage-liquid phase method.The target product was purified by reversed-phase high-performanced liquid chromatography(RP-HPLC).RESULTS The purity of crude linaclotide was 75.4%.After the purification,the purity reached to 99.32%.The product was characterized by high resolution mass spectrometry(HR-MS),and its chromatographic characteristics were consistent with those of the standard.CONCLUSION With this method,large-scale preparation of linaclotide can be achieved simply and efficiently,which provides references for industrial production.
作者
付玉清
李新宇
姚林
岳泽乐
FU Yu-qing;LI Xin-yu;YAO Lin;Yue Ze-le(Shenzhen JYMed Technology Co.,Ltd.,Shenzhen 518118,China)
出处
《海峡药学》
2020年第10期32-35,共4页
Strait Pharmaceutical Journal
