文献类型：专利

英文题名：Preparing linaclotide, by connecting side chain protected amino acid with Wang resin, adding e.g. organic base to flask, adding dimethylformamide, coupling resin to amino acid sequence, and adding iodine into sodium phosphate buffer

作者：LIU B;WU J;TIAN H;WANG Q;ZOU R

机构：[1]UNIV EAST CHINA SCI & TECHNOLOGY

申请号：CN104628826-A

申请日：2013-11-11

公开日：2015-05-20

语种：英文

收录：DERWENT

摘要：NOVELTY - Preparing linaclotide (I) involves connecting a side chain protected amino acid with Wang resin, adding organic base and coupling agent to flask, adding dimethylformamide (DMF) to reaction solution, conjugating obtained product with Fmoc-cysteine, performing above-mentioned steps, coupling resin to amino acid sequence, cutting amino acid sequence conjugated to resin to obtain a compound HO-Tyr-Cys-Gly-Thr-Cys-Ala-Pro-Asn-Cys-Cys-Tyr-Glu-Cys-Cys-H, and adding elemental iodine into a sodium phosphate buffer solution to carry out cysteine oxidation. USE - The method is useful for preparing linaclotide (claimed). ADVANTAGE - The method is simple, and carried out under mild reaction conditions, provides high yield of pure linaclotide, and increases final product yield by 30-60%. DETAILED DESCRIPTION - Preparing linaclotide of formula (I) involves (a) utilizing a standard fluorenylmethyloxycarbonyl (Fmoc) technology connecting a side chain protected amino acid with Wang resin in a reaction flask, adding organic base and coupling agent to the flask, adding dimethylformamide (DMF) to the reaction solution, and removing the Fmoc protecting group to obtain a protective group associated with tyrosine resin, (b) conjugating obtained product with Fmoc-cysteine in a reaction flask, adding organic base and coupling agent to the flask, adding DMF to the reaction solution, and removing the Fmoc protecting group to obtain a protective group associated with cysteine resin, (c) successively repeating step (b), where amino acid sequence is replaced with Fmoc-glycine, Fmoc-threonine, Fmoc-alanine, Fmoc-proline, Fmoc-asparagine, Fmoc-cysteine, and Fmoc-glutamic acid, removing the Fmoc protecting group, and sequentially coupling resin to amino acid sequence, (d) cutting amino acid sequence conjugated to the resin to obtain a compound HO-Tyr-Cys-Gly-Thr-Cys-Ala-Pro-Asn-Cys-Cys-Tyr-Glu-Cys-Cys-H, and (e) adding elemental iodine into a sodium phosphate buffer solution to carry out cysteine oxidation.