Patent ID: 7524813
Filing Date: 2009-04-28
Classification: A61K,C07C,C07K,C12P

Abstract:
1. A conjugated peptide obtainable by a method comprising the steps of i) reacting in one or more steps a peptide with a first compound bearing one or more functional groups, which are not accessible in any of the amino acids residues constituting said peptide, in the presence of an enzyme capable of catalyzing the incorporation of said first compound into the C-terminal of said peptide to form a transacylated peptide, and ii) reacting in one or more steps said transacylated peptide with a second compound comprising one or more functional groups, wherein said functional group(s) do not react with functional groups accessible in the amino acid residues constituting said peptide, and wherein said functional group(s) in said second compound is capable of reacting with said functional group(s) in said first compound so that one or more covalent bond between said transacylated peptide and said second compound is formed, and wherein said peptide P is reacted in one or more steps with a first compound, which is an α-amino acid amide represented by the formula in the presence of carboxypeptidase to form a transacylated peptide of the formula said transacylated peptide being further reacted in one or more steps with a second compound of the formula to form a conjugated peptide of the formula wherein R represents a linker or a bond; P′ represents a peptide formed by removing the C-terminal amino acid from the peptide P, P selected from the group consisting of: human insulin, GLP-1, GLP-2, human growth hormone (GH) and factor VII; X is selected from amongst keto-, aldehyde-, —NH—NH Y is selected from amongst keto-, aldehyde-, —NH—NH E represents a linker or a bond; A represents oxime, hydrazones, phenylhydrazone, semicarbazone or triazole moieties; and Z comprises the moiety to be conjugated to the peptide where said moiety decreases the clearance of compound of the formula in comparison with the clearance of peptide P and where the moiety is selected from the group consisting of: PEG; mPEG; straight, branched and/or cyclic C 2-22 alkyl, C 2-22 alkenyl, C 2-22 alkynyl, C 1-22 heteroalkyl, C 2-22 heteroalkenyl, C 2-22 heteroalkynyl, wherein each may be optionally substituted with one or more substituents selected from hydroxyl, halogen, carboxyl, heteroaryl and aryl; steroid; lipids; polysaccharides; ethylene/maleic anhydride polymer; and peptides.