NOVEL PROINSULIN GLARGINE AND METHOD FOR PREPARING INSULIN GLARGINE THEREFROM

    公开(公告)号:EP4206220A1

    公开(公告)日:2023-07-05

    申请号:EP20953153.2

    申请日:2020-12-24

    IPC分类号: C07K14/62 C12N15/11

    摘要: The present invention discloses novel proinsulin glargine and a method for preparing insulin glargine therefrom, and belongs to the technical field of recombinant protein preparation. According to the present invention, a sequence of the proinsulin glargine containing SOD fusion peptide subjected to site-directed mutagenesis and "0 C peptide" is designed; recombinant Escherichia coli engineering bacteria for expressing insulin glargine are constructed; insulin glargine fusion protein in a form of an inclusion body is expressed by inducing the engineering bacteria; and denaturation, renaturation, modification, enzyme digestion, separation and purification are carried out to obtain a mature insulin glargine active pharmaceutical ingredient. According to the present invention, the SOD fusion peptide sequence is mutated to enhance the fermentation yield of the insulin glargine by 75%; and a "0 C peptide" strategy is adopted to avoid remaining of C-peptide residues and reduce the quality loss and miscleavage impurities in the enzyme digestion transformation. The purity of the insulin glargine active pharmaceutical ingredient prepared in the present invention is up to 99.9%, and the maximum single impurity content is controlled at 0.05%.