METHODS FOR AMINO GROUP PEGYLATION TO OBTAIN SITE-SPECIFIC CONJUGATES OF PROTEINS Russian patent published in 2020 - IPC A61K31/00 A61K38/28 A61K47/60 C07K14/62 

FIELD: medicine.

SUBSTANCE: group of inventions relates to a method of producing a protein-polyethylene glycol conjugate. Method comprises preparing an aqueous protein solution containing protein, a buffer solution which provides the desired pH, and a chelating agent, wherein said protein is insulin, said buffer solution, providing the desired pH, is an acetate or citrate, and said chelating agent is selected from a group consisting of aminopolycarboxylic acid, hydroxyaminocarboxylic acid, N-substituted glycine, 2-(2-amino-2-oxoethyl)aminoethanesulfonic acid (BES) and deferoxamine (DEF); adding to the aqueous protein solution a boron-containing reducing agent and methoxypolyethylene glycol aldehyde, wherein the boron-containing reducing agent and methoxypolyethylene glycol aldehyde are taken in molar ratio from 25:1 to 1.5:1; and reacting methoxy polyethylene glycol aldehyde and protein to form a protein-polyethylene glycol conjugate, wherein the buffer solution during reaction provides pH of the aqueous protein solution from 3.88 to 4.27, and as a result of said interaction of methoxy polyethylene glycol aldehyde with said protein conjugate of PEG and protein, pegylated in one position is formed. Group of inventions also relates to a method of producing an insulin-polyethylene glycol conjugate and a method of producing delayed release microspheres containing a protein conjugate with polyethylene glycol.

EFFECT: group of inventions enables to obtain pegylated proteins with high output, as well as higher clinical effectiveness in diabetes and other diseases.

35 cl, 16 dwg, 19 ex