RU2002135041A - METHOD FOR PRODUCING A HUMAN INSULIN COMBINED DRUG - Google Patents

Claims (1)

A method of obtaining a combined preparation of human insulin, which includes obtaining human insulin ester by transpeptidation of porcine insulin with an excess of threonine di-tert-butyl ether in an aqueous-organic medium in the presence of trypsin, chromatographic purification of the obtained insulin ester, deprotection, crystallization of human insulin, preparation of a portion of short the action of the combined drug by dissolving a certain amount of insulin crystals in dilute acid and mixing with solutions preservative, isotonic agent and substances with a buffer capacity, preparation of a part of the prolonged action of the combined preparation by dissolving the remaining insulin crystals also in dilute acid, followed by the addition of acidic solutions of zinc ions and protamine sulfate, mixing with buffered solutions of m-cresol, phenol and glycerol, keeping the solution until the formation of crystals and the connection of the obtained parts of the combined insulin preparation, characterized in that the transpeptidation reaction is carried out dyat with a molar excess of threonine di-tert-butyl ether from more than 100 to 500 and with a weight ratio of trypsin and pork insulin from 1 to 300-1000, then the reaction mixture is diluted with water 2-3 times, purification by high-pressure liquid chromatography of insulin ester of man and obtained after direct crystallization of human insulin, in the preparation of both parts of the combined preparation, the dissolution of insulin crystals in dilute acid is carried out in stages, and a fine suspension of ins crystals is first obtained the uline in water, and then dilute acid is added to it, and when preparing a part of the prolonged action of the combined preparation, the mixture obtained after mixing with buffered solutions of m-cresol, phenol and glycerol is kept at a temperature of 18-21 ° C for 20-22 h .