METHOD FOR PREPARING READY MEDICINAL FORMULATION OF INSULIN WITH SHORT EFFECT Russian patent published in 2005 - IPC

FIELD: medicine, endocrinology, chemical-pharmaceutical industry, hormones.

SUBSTANCE: method involves preparing human insulin ester by transpeptidation reaction of porcine insulin in the molar excess of threonine di-tert.-butyl ester in an aqueous-organic medium in the presence of trypsin, inhibition of reaction by acidification, purification of preparing human insulin ester by chromatography, removal of protecting groups with trifluoroacetic acid and purification of prepared human crude insulin, its following dissolving in diluted acid and mixing with preserving agent solutions, isotonic agent and substances with buffer capacity. Preparing human insulin ester is carried out in the weight ratio trypsin to porcine insulin = 1:(300-1000) followed by additional inhibition of reaction by dilution of the reaction mixture with water by 2-3 times before acidifying. Purification of human insulin ester is carried out by HPLC method followed by precipitation of ester derivative fractions in the presence of zinc ions, removal of protecting groups and preparing human crude insulin crystals that is purified by repeated carrying out HPLC method. Both processes of purification are carried out by using sorbent DIASOGEL ODS (C18) as immobile phase with particles size from 15 mcm and pores size from 100 to 150 . At the first stage 0.06 M glycine - HCl buffer containing 0.015 M of ammonium sulfate and propanol-2 in the concentration from 20% to 35%, pH 2.5 is used as a mobile phase, and 0.05 M acetate buffer with the content of propanol-2 from 15% to 25%, pH 2.5 is used at the second stage. The purified human insulin is dissolved in diluted acid by stages wherein finely dispersed suspension of insulin crystals in water is prepared followed by addition a diluted acid to it. Invention provides the development of effective, economy method for preparing the ready medicinal formulation of insulin with short effect and with low immunological properties that provides reducing loss of insulin in the process of its preparing. Invention can be used in manufacturing ready insulin formulations of high quality and with short effect.

EFFECT: improved preparing method.

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