Dimethyl Sulfoxide → (-)-Asparagine

In a 1.5-mL Eppendorf tube, DIPEA (4.8 μL, 27.6 μmol) and the 0.03 M sugar linker reagent prepared above (15.0 μL, 4.5 μmol) were added to a DMSO (300 μL) solution of Lys26Arg34GLP-1(7-37) synthesized in Synthesis Example 4 (SEQ ID NO: 80) (2.8 mg, 0.83 μmol). After stirring at room temperature for 2 hours, the reaction was terminated by the addition of an aqueous solution (200 μL) of glycine (2 mg, 26.6 μmol). Fractions of peaks at a retention time of 15.5 min were directly collected by HPLC [column: Zorbax 300SB-CN, φ4.6×250 mm, gradient: A solution: 0.1% TFA water, B solution: 0.09% TFA/10% water/90% AN 1.0 ml/min; 10→40% B (0-8 min) 40-+50% B (8-20 min) linear gradient). After freeze-drying, the following 26Lys-asialo oligosaccharide chain Asn linker-modified Arg34GLP-1(7-37) (0.7 mg) was obtained (MALDI TOF Mass calculated for [M (average)+H]+ 5236.35. found 5236.1):

1. customsynthesized in Synthesis Example 4 (SEQ ID NO
2. customFractions of peaks at a retention time of 15.5 min were directly collected by HPLC [column
3. custom10→40% B (0-8 min) 40-+50% B (8-20 min) linear gradient
4. customAfter freeze-drying

ORD record

ord-a0507a51c40743c2b8c9a484e850b8da

Source dataset

uspto-grants-2014_07

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US08765669B2