Title:
NOVEL PROINSULIN GLARGINE AND METHOD FOR PREPARING INSULIN GLARGINE THEREFROM
Document Type and Number:
WIPO Patent Application WO/2022/052368
Kind Code:
A1
Abstract:
The present invention belongs to the technical field of recombinant protein preparation, and discloses a novel proinsulin glargine and a method for preparing insulin glargine therefrom. In the present invention, a sequence of proinsulin glargine containing an SOD fusion peptide that has undergone site-directed mutation and "0C peptide" is designed, a recombinant E. coli engineered bacteria expressing insulin glargine is constructed, and the engineered bacteria is induced to express an insulin glargine fusion protein in the form of inclusion bodies, then same undergoes denaturation, renaturation, modification, enzyme cleavage, separation, and purification to obtain a mature insulin glargine active ingredient. In the present invention, by means of mutating an SOD fusion peptide sequence, the fermentation yield of insulin glargine is increased by 75%. By means of adopting a "0C peptide" strategy, the remnants of C-peptide residues is avoided, and the quality loss and mis-cutting impurities of enzyme cleavage conversion are reduced. The purity of the insulin glargine active ingredient prepared by the invention is up to 99.9%, and the maximum single impurities are controlled at 0.05%.
Inventors:
TANG CHUANGEN (CN)
WANG JING (CN)
PAN SHANGSHU (CN)
FAN XIAOYANG (CN)
LIU XIAORUI (CN)
CHEN SONG (CN)
ZHANG HAONING (CN)
WANG JING (CN)
PAN SHANGSHU (CN)
FAN XIAOYANG (CN)
LIU XIAORUI (CN)
CHEN SONG (CN)
ZHANG HAONING (CN)
Application Number:
PCT/CN2020/138950
Publication Date:
March 17, 2022
Filing Date:
December 24, 2020
Export Citation:
Assignee:
AMPHASTAR NANJING PHARMACEUTICALS INC (CN)
International Classes:
C07K14/62; C12N15/11
Foreign References:
| CN103981242A | 2014-08-13 | |||
| CN102015762A | 2011-04-13 | |||
| US5656722A | 1997-08-12 | |||
| CN103981242A | 2014-08-13 | |||
| US6777207B2 | 2004-08-17 |
Other References:
BERGER M, A COMMENT. DIABETES RES CLIN PRACT, vol. 6, 1989, pages 25 - 31
SANLIOGLU AD ET AL.: "Clinical utility of insulin and insulin analogs", ISLETS, vol. 5, no. 2, 2013, pages 67 - 78
FRANK, B.H ET AL.: "Pierce Chemical Company", ROCKFORD, 1981, pages 729 - 738
See also references of EP 4206220A4
SANLIOGLU AD ET AL.: "Clinical utility of insulin and insulin analogs", ISLETS, vol. 5, no. 2, 2013, pages 67 - 78
FRANK, B.H ET AL.: "Pierce Chemical Company", ROCKFORD, 1981, pages 729 - 738
See also references of EP 4206220A4
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