METHOD FOR PRODUCING SUBSTANCE OF INSULIN SUITABLE FOR DELAYED RELEASE DOSAGE FORM

摘要

The method for producing the substance of insulin suitable for delayed release dosage form comprises the aqueous and alcohol extraction of the porcinepancreas followed by concentration and isolation of insulin substance from the extract. Then the substance is purified chromatographically with the aim of discarding proinsulin, the foreign proteins, and the insulin-related impurities. The product is crystallized. The substance isolated is dissolved in the acid and mixed with the aqueous solutions of other ingredients. The porcine insulin substance is subjected to the reversed-phase chromatography on the silica gels C8 or C18. With the aim of purifying the substance from proinsulin and the foreign proteins the elution is performed employing aqueous-organic solution with pH not exceeding 3.5 comprising 12-21% of isopropanol or propanol (preferably 18-20%) and no more than 0.25 M sodium acetate, ammonia acetate, ammonia sulfate, or sodium sulfate (preferably 0.01-0.06 M). The substance is purified from the insulin-related impurities employing isocratic mode of elution or the gradient of the organic modifier. The substance of insulin is dissolved in hydrochloric acid with the addition of zinc chloride or zinc oxide. Then the aqueous solution of sodium acetate and sodium chloride is prepared. Both solutions are mixed and the seed for priming crystallization is added. The reaction mixture is stabilized with the aqueous solution of zinc chloride or zinc oxide. The citric acid and the preserving agent are also added.