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A kind of preparation method of insulin glargine crystal

A technology of insulin glargine and crystal, which is applied in the field of insulin crystallization, can solve the problems of analysis, many interfering impurities and low purity, etc.

Active Publication Date: 2022-04-26
YICHANG HEC CHANGJIANG PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the reports on the crystallization methods of insulin glargine in the prior art are all crystallization methods used in the production of insulin glargine. Most of the obtained products are in the form of mixed crystals, with low purity and many interfering impurities, which cannot be used for crystal structure analysis. analyze

Method used

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  • A kind of preparation method of insulin glargine crystal
  • A kind of preparation method of insulin glargine crystal
  • A kind of preparation method of insulin glargine crystal

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0032] This example provides a method for preparing insulin glargine crystals.

[0033] 1. Preparation of stock solution

[0034] 1M citric acid-1M Tris buffer solution: Weigh 19.2g of anhydrous citric acid and 12.1g of Tris, add 100ml of water to dissolve, adjust the pH with NaOH solution, and obtain a buffer solution with a pH value of 8.0-11.0.

[0035] 1M ammonium acetate solution: weigh 0.77g of ammonium acetate, add 10ml of water to dissolve it.

[0036] 4MNaOH: Weigh 1.16g of sodium hydroxide, add 5ml of water to dissolve.

[0037] 1M phenol solution: weigh 94mg of phenol, add 1ml of water to dissolve, and mix well.

[0038] 0.5M zinc acetate solution: Weigh 219mg of zinc acetate, add 2ml of water to dissolve, and mix well.

[0039] 0.01M hydrochloric acid solution: Measure 0.9ml hydrochloric acid, dilute it with water to 1000ml, and shake well.

[0040] 2. Preparation of insulin glargine solution

[0041] Weigh 20mg of insulin glargine, add 5ml of 0.01M hydrochlor...

Embodiment 2

[0060] This example provides a method for preparing insulin glargine crystals. Compared with Example 1, the only difference is that the composition of the crystallization solution is: Tris 0.4M, citric acid 0.4M, ammonium acetate 25mM; the pH of the crystallization solution is 8.5.

[0061] In this embodiment, insulin glargine crystals in the form of single crystals can be obtained.

Embodiment 3

[0063] This example provides a method for preparing insulin glargine crystals. Compared with Example 1, the only difference is that the composition of the crystallization solution is: Tris 0.7M, citric acid 0.7M, ammonium acetate 30mM; the pH of the crystallization solution is 9.0.

[0064] In this embodiment, insulin glargine crystals in the form of single crystals can be obtained.

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Abstract

The invention relates to the technical field of insulin crystallization, in particular to a method for preparing insulin glargine crystals. The method for preparing insulin glargine crystals adopts the gas-phase diffusion hanging drop method, comprising the following steps: (1) preparing a crystallization solution containing the following components: Tris 0.2-1.0M, citric acid 0.2-1.0M; the pH of the crystallization solution is 8.0 to 11.0; (2) Mix the insulin glargine solution with the crystallization solution to obtain a mixed crystallization solution; (3) Hang the mixed crystallization solution above the mother liquor pool filled with the crystallization solution, and let it stand for cultivation . The method can be used to obtain insulin glargine crystals with regular shape and suitable size in the form of single crystals, and the resolution of X-ray diffraction is high, and the crystal structure of insulin glargine can be correctly analyzed.

Description

technical field [0001] The invention relates to the technical field of insulin crystallization, in particular 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 glycine-arg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/62C07K1/30
CPCC07K14/62C07K2299/00
Inventor 郭林峰张淼封启媛徐军林小鹊章琛莫隆兴李晓平陈小锋李文佳
Owner YICHANG HEC CHANGJIANG PHARMA CO LTD
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