Patent ID: 9725493
Date: 2017-08-08
CPC Classifications: A61K,A61P,C07K,C12P

Claim:
1. A method for preparing a des-tripeptide [B1-B3] insulin analogue, the method comprising: providing a folded single-chain or two-chain insulin-related precursor polypeptide containing Arg or Lys at position B3; adding trypsin to the folded single-chain or two-chain insulin-related precursor polypeptide under conditions conducive to the proteolytic activity of trypsin to provide a reaction mixture; incubating the reaction mixture for sufficient time to yield a des-tripeptide[B1-B3]-des-octapeptide [B23-B30]-insulin intermediate product; adding a synthetic peptide in molar excess, wherein the synthetic peptide comprises residues corresponding to residues B23-B30 of insulin or an analogue thereof; adding trypsin under conditions conducive to the enzymatic activity of trypsin such that trypsin catalyzes formation of a peptide bond between ArgB22 of the des-tripeptide [B1-B3]-des-octapeptide [B23-B30]-insulin intermediate product and the synthetic peptide to yield the des-[B1-B3]insulin analogue.