3 results about "Insulin Precursor" patented technology

The invention provides a preparation method and application of a long-acting insulin analogue, and the preparation method comprises the following steps: taking eicosandioic acid A as a starting raw material, carrying out unilateral esterification with trimethylsilyl ethanol to obtain an intermediate B, reacting the intermediate B with Hosu to obtain an active intermediate C, reacting the intermediate C with L-glutamic acid-1-trimethylsilyl ethyl ester to obtain an intermediate D, and carrying out condensation reaction on the intermediate D to obtain the long-acting insulin analogue. The intermediate D reacts with Hosu to obtain an active intermediate E, the intermediate E reacts with AEEA-AEA to obtain an intermediate F, the intermediate F reacts with Hosu to obtain an active intermediate G, and trimethylsilylethyl is removed from the intermediate G through tetrabutylammonium fluoride trihydrate to obtain the Ecobainsulin side chain. The route of the method has the following advantages that related raw materials are simple and easy to obtain, the reaction condition is mild, the yield is high, and the method is suitable for industrial production. The preparation method is simple and easy to implement, large-scale production is easy, the prepared side chain of the Ekoinsulin and A14E, B16E, B25H and DesB30 insulin precursors are coupled with lysine at the B29 site under the alkaline condition to obtain a crude Ekoinsulin product, and the crude product is subjected to two-step purification to obtain the Ekoinsulin with the purity of 99.0% or above.

The invention provides a preparation method of an Icodec insulin analogue precursor, which belongs to the technical field of biology and comprises the following steps: (1) construction and culture of engineering bacteria: inserting an expression vector into a base sequence of a fusion protein containing the Icodec insulin precursor, recombining the expression vector into yeast or escherichia coli to obtain the engineering bacteria, and performing fermentation and induced expression to obtain the Icodec insulin analogue precursor; fermentation liquor or thalli containing the Icodec insulin precursor fusion protein are obtained; (2) carrying out chromatographic purification treatment on the fermentation liquor of the yeast engineering bacteria, and then carrying out enzyme digestion; thalli obtained by fermenting escherichia coli engineering bacteria need to be crushed, subjected to denaturation and renaturation and then subjected to enzyme digestion; (3) reverse phase chromatography purification: carrying out reverse phase chromatography purification treatment on the enzyme digestion reaction solution by using C8 silica gel resin to obtain a reverse phase chromatography solution; and (4) precipitate collection: diluting the reverse phase chromatography solution, adjusting the pH value, and centrifugally collecting the precipitate. The method has the advantages of few chromatography processes, simple process and high recovery rate.

The present invention relates to a recombinant insulin precursor. To improve the recombinant production of insulin, a recombinant insulin precursor according to the chemical formula LBCA is provided, wherein A is the A-chain of insulin, B is the B-chain of insulin, C is a C-peptide connecting the A-chain and the B-chain, and L is an N-terminal L-peptide. The C-peptide comprises an amino acid sequence according to SEQ ID NO: 1 and / or the L-peptide comprises an amino acid sequence according to SEQ ID NO: 2.