Patent ID: 7402565
Filing Date: 2008-07-22
Classification: C07K,C12P

Abstract:
1. A method for making an acylated insulin or an insulin analogue, said method comprising: a) acylating an insulin intermediate with an acylating agent in a solvent, wherein said insulin intermediate has the formula: wherein the X 3 —X 4 sequence is selected from the group of sequences consisting of Asp-Pro, Glu-Pro, Asp-Glu, Asp-Gly and Glu-Gly; and the X 1 —X 2 sequence is selected from the group of sequences consisting of Asp-Asp-Gly-Asp-Pro (amino acids 1-5 of SEQ ID NO:1), Glu-Glu-Gly-Glu-Pro (amino acids 1-5 of SEQ ID NO:2), Asp-Asp-Gly-Asp-Gly (amino acids 1-5 of SEQ ID NO:3) and Glu-Glu-Gly-Glu-Gly (amino acids 1-5 of SEQ ID NO:4), B-chain is the B-chain of human insulin or an analogue thereof, and A-chain is the A-chain of human insulin or an analogue thereof, at the C-terminal lysine amino group in the B-chain, and b) cleaving the peptide bond between Arg and B1 and between Arg and A1 in the acylated intermediate by trypsin or a trypsin like protease to produce an acylated insulin or insulin analogue.