METHOD FOR PURIFYING SOLID-PHASE SYNTHETIC CRUDE LIRAGLUTIDE

摘要

The present invention relates to the field of biomedicine, and in particular, to a method for purifying solid-phase synthetic crude liraglutide. The method comprises: dissolving solid-phase synthetic crude liraglutide in an aqueous acetonitrile solution to obtain a crude peptide solution; and obtaining liraglutide with high purity and high yield through four-step HPLC purification.

主权项

1. A method for purifying crude liraglutide obtained from solid-phase synthesis, which is characterized by comprising the following steps:
(a) a solution of crude liraglutide is obtained by dissolving crude liraglutide obtained from solid-phase synthesis in aqueous acetonitrile solution; (b) the solution of crude liraglutide is subjected to a first HPLC purification using octylsilane bonded silica as stationary phase, and using aqueous isopropanol solution containing 0.1-0.2% trifluoroacetic acid as mobile phase A and acetonitrile containing 0.1-0.2% trifluoroacetic acid as mobile phase B eluting at a linear gradient from 20-40% B to 40-60% B, and target peak is collected as the first fraction; (c) the first fraction is subjected to a second HPLC purification using cyanosilane bonded silica as stationary phase, and using 0.05-0.15% (mass concentration) aqueous perchloric acid solution as mobile phase A and 0.05-0.15% (mass concentration) perchloric acid in acetonitrile as mobile phase B eluting at a linear gradient from 40% B to 70% B, and target peak is collected as the second fraction; (d) the second fraction is subjected to a third HPLC purification using octylsilane bonded silica as stationary phase, and using 0.01-0.06% (mass concentration) aqueous ammonia solution as mobile phase A and acetonitrile of chromatographic grade as mobile phase B eluting at a linear gradient from 30% B to 60% B, and target peak is collected as the third fraction; and (e) purified liraglutide is obtained from the third fraction by rotatory evaporation under reduced presser and lyophilization.