Facile synthesis of insulin fusion derivatives through sortase A ligation.
10.1016/j.apsb.2020.11.011
- Author:
Maria M DISOTUAR
1
;
Jake A SMITH
1
;
Jinze LI
1
;
Steve ALAM
1
;
Nai-Pin LIN
1
;
Danny Hung-Chieh CHOU
1
Author Information
1. Department of Biochemistry, School of Medicine, University of Utah, Salt Lake City, UT 84112, USA.
- Publication Type:Journal Article
- Keywords:
Alb, albumin;
Albumin-binding peptide SA21;
Boc, tert-butyloxycarbonyl;
DCM, dichloromethane;
DIEA, N,N-diisopropylethylamine;
DMEM, Dulbecco's Modified Eagle Medium;
DMF, dimethylformamide;
DMSO, dimethyl sulfoxide;
DOI, desoctapeptide (B23−30) insulin;
Diabetes mellitus;
EDT, 1,2-ethanedithiol;
FBS, fetal bovine serum;
Fmoc, 9-fluorenylmethoxycarbonyl;
HATU, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate;
HBTU, O-(benxontriazol-1-yl)-1,1,3,3-tetramethyluronium;
HPLC, high performance liquid chromatography;
HTRF, homogeneous time resolved fluorescence;
IR-B, human insulin receptor isoform B;
ITT, insulin tolerance test;
Insulin synthesis;
LC‒MS, liquid chromatography mass spectrometry;
Long-acting insulin;
Mtt, 4-methyltrityl;
NBD-X, 6-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoic acid;
STZ, streptozotocin;
Sortase A (SrtA) ligation;
SrtA, sortase A;
THF, triflouroacetic acid;
TIS, triisoproylsilane;
i.p., intraperitoneal;
pAkt, phosphorylated protein kinase B;
t-Bu, tert-butyl
- From:
Acta Pharmaceutica Sinica B
2021;11(9):2719-2725
- CountryChina
- Language:English
-
Abstract:
Insulin derivatives such as insulin detemir and insulin degludec are U.S. Food and Drug Administration (FDA)-approved long-acting insulin currently used by millions of people with diabetes. These derivatives are modified in C-terminal B29 lysine to retain insulin bioactivity. New and efficient methods for facile synthesis of insulin derivatives may lead to new discovery of therapeutic insulin. Herein, we report a new method using sortase A (SrtA)-mediated ligation for the synthesis of insulin derivatives with high efficiency and functional group tolerance in the C-terminal B chain. This new insulin molecule (Ins-SA) with an SrtA-recognizing motif can be conjugated to diverse groups with N-terminal oligoglycines to generate new insulin derivatives. We further demonstrated that a new insulin derivative synthesized by this SrtA-mediated ligation shows strong cellular and
