METHOD FOR PREPARING COMBINED DOSAGE FORM OF HUMAN INSULIN

摘要

The method for preparing the combined dosage form of human insulin comprises transpeptidation of porcine insulin with the excess of di-tret-butyl ester of threonine in aqueous-organic medium in the presence of trypsin for obtaining the ether of the human insulin. Transpeptidation is performed in 100-500-molar excess of di-tret-butyl ester of threonine and 1 : (300-1000) mass ration of trypsin to porcine insulin. The reaction mixture is diluted 2-3-fold with the water. The insulin ether and insulin obtained upon the direct crystallization are purified using high-performance liquid chromatography. When the short-term and long-term components of the combined dosage form of human insulin are prepared, the insulin crystals are dissolved in the diluted acid step-by-step. First, the fine dispersion of insulin crystals in the water obtained and then the diluted acid is added. To prepare the long-term components of the combined dosage form, the mixture with the buffered solutions of m-cresol, phenol, and glycerol is kept at the temperature of 18-21 DEGREE C for 20-22 hours.