A method for preparing insulin glargine crystals

Abstract

The invention relates to a method for preparing a recombinant glycine-arginine insulin crystal. The method specifically comprises the following steps of (1) dissolving recombinant glycine-arginine insulin, a phenol derivative, a zinc-containing substance and organic acid in water so as to obtain crystal fluid; and (2) taking a pH regulating agent for regulating the pH value of the crystal fluid to 4.5-7.0, performing stirring for 3-9 hours, and performing separation so as to obtain the recombinant glycine-arginine insulin crystal, wherein the concentration of the recombinant glycine-arginine insulin in the crystal fluid is 1-3.5g / L, the mass / volume percentage of the phenol derivative is 0.01-1.00%, the mass / volume percentage of the zinc-containing substance is 0.005-1.5%, and the mass / volume percentage of the organic acid is 0.01-2.0%. The crystal fluid prepared by the method does not contain organic solvents, the operation process is safe, the technology is simple and easy to control, poisonous and harmful substances in chromatography in the previous step can be effectively removed, the time for filtration, collection, crystal washing and freeze drying of products is shortened, the cost is reduced, and the method is suitable for industrialized production of the recombinant glycine-arginine insulin.

Description

technical field [0001] The present invention relates to the field of crystallization of insulin. Specifically relates to a method for preparing insulin glargine crystals. Background technique [0002] Insulin is a protein hormone secreted by pancreatic β cells stimulated by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, and glucagon. It was first discovered by Canadians F.G. Banting and C.H. Best in 1921. It began to be used clinically in 1922 to save the diabetic patients who died in the past. In recent years, the prevalence of diabetes in the world has been increasing rapidly, and the demand for insulin is increasing. Simple extraction from animal cells can no longer meet the needs of patients, and the production of insulin and insulin analogues is imminent. One of the newly developed long-acting insulin analogues, insulin glargine, is being increasingly accepted and used by doctors and patients. [0003] Insulin glargine (recombinant gl...

AI Technical Summary

Problems solved by technology

[0006] Chinese patent CN102219851B discloses a method for preparing insulin glargine crystals from a crystallization solution containing insulin glargine, organic solvents, zinc, phenol derivatives, salts, and organic acids under the condition of pH 7.0 to 9.0. The prepared crystal has the advantages of stable solid molecular form, easy mass production operation, small sedimentation volume, easy separation from the supernatant, short centrifugation time and freeze-drying time, and high production efficiency. However, there are a large number of organic reagents in the crystallization process. Residues of poor and highly toxic organic solvents will seriously affect the quality of insulin products

This will not only bring risks to the final crystallized product, but also bring a lot of trouble to subsequent operations

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