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Method for synthesizing and preserving insulin dimer crystal

A synthesis method and insulin technology, applied in the preparation methods of insulin and peptides, chemical instruments and methods, etc., can solve the problems of complex preparation methods of insulin dimers, short storage time, unstable products, etc., and achieve stable and easy storage methods. performance, stable preparation conditions and high purity

Inactive Publication Date: 2014-12-10
韩飞 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims to solve the technical problems of complex preparation methods, low yield, unstable product, and short storage time of existing insulin dimers; and provides a method for synthesizing and storing insulin dimer crystals

Method used

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  • Method for synthesizing and preserving insulin dimer crystal
  • Method for synthesizing and preserving insulin dimer crystal
  • Method for synthesizing and preserving insulin dimer crystal

Examples

Experimental program
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specific Embodiment approach 1

[0024] Specific embodiment one: the synthesis method of insulin dimer crystal in this embodiment is carried out according to the following steps:

[0025] Step 1, arginine, glutamic acid, L-glutamine, methionine, L-glutathione, zinc acetate, sodium citrate, tris (Tris), sodium sulfate, Add 2-N-morpholineethanesulfonic acid (MES) and phenol acetone solution into deionized water to configure the reaction solution, and adjust the pH value;

[0026] Step 2. Stir 200ml of the prepared reaction solution at 12°C for 2h, add 80mL of R with a concentration of 32g / L 6 Insulin hexamer aqueous solution (prepared with deionized water), mix well, add 20ml of dipeptide with a concentration of 0.1mol / L, continue to stir for 2h, then add 10ml of catalyst, and then continue to stir and react at 12°C for 12 hours , the stirring speed is kept at 300r / min (using a magnetic stirrer) throughout the reaction process to obtain an insulin dimer solution;

[0027] Described catalyst is made up of argi...

specific Embodiment approach 2

[0042] Embodiment 2: This embodiment differs from Embodiment 1 in that: in step 3, the pH of the crystallization solution is adjusted to 6-6.8, and other reactions remain unchanged, and the product is stored in a sealed light-proof glass container at 3-10°C.

[0043] Insulin dimers can be stored for more than 1 year.

specific Embodiment approach 3

[0044] Specific embodiment three: the synthesis method of insulin dimer crystal in this embodiment is carried out according to the following steps:

[0045] The synthetic method of insulin dimer crystal in this embodiment is carried out according to the following steps:

[0046] Step 1, arginine, glutamic acid, L-glutamine, methionine, L-glutathione, zinc acetate, sodium citrate, tris (Tris), sodium sulfate, Add 2-N-morpholineethanesulfonic acid (MES) and phenol acetone solution into deionized water to configure the reaction solution, and adjust the pH value;

[0047] Step 2. Stir 200ml of the prepared reaction solution at 14°C for 1.5h, add 80mL of T3R3 insulin hexamer aqueous solution with a concentration of 32g / L, mix well, add 20ml of dipeptide with a concentration of 0.1mol / L, and continue Stir for 2 hours, then add 10ml of catalyst, and then continue to stir and react at 14°C for 12 hours. During the reaction, the stirring speed is always kept at 600r / min (using magneti...

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Abstract

The invention discloses a method for synthesizing and preserving an insulin dimer crystal, relating to a method for synthesizing and preserving insulin and aiming at solving the technical problems of complexity in method, low yield, unstable products and short storage time in the existing methods for preparing insulin dimmers. The synthesis process comprises the steps of enabling insulin hexamer as the raw material to react with dipeptide in the presence of a catalyst, and then crystallizing in a crystallization solution prepared from arginine, glutamic acid, L-glutamine, L methionine, L-glutathione, zinc acetate, sodium citrate, trihydroxymethyl aminomethane, sodium chloride, 2-N-morpholinoethanesulfonic acid and a phenol propanol solution; and the preserving process comprises the steps of putting the insulin dimer crystal in the crystallization solution with the pH value of 6-6.8 and storing in a closed glass container away from light at 3-10 DEG C. The insulin dimer prepared according to the method is a natural structure, has no genetically engineered components and small side effects, is suitable for people who are allergic to insulin analogues, and can be stored for more than one year.

Description

technical field [0001] The invention relates to a method for synthesizing and preserving insulin. Background technique [0002] As an important protein, insulin has been studied for nearly one hundred years. Insulin is essentially a biologically active protein, usually in the form of a relatively stable hexamer, and is widely used in the treatment of diabetes. [0003] The current insulin preparations can be divided into five categories according to the time of action, among which insulin aspart and insulin lispro have the fastest onset of action, and the onset time is about 15 minutes. The disadvantage is that these insulin drugs are all insulin analogs, and their structure is different from that of natural insulin. The corresponding amino acids are used to replace the original amino acids to achieve the purpose of rapid onset of action. It is not suitable for diabetic patients who are allergic to the corresponding amino acids or insulin analogs , and synthetic insulin an...

Claims

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Application Information

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IPC IPC(8): C07K14/62C07K1/30
CPCC07K14/62C07K2299/00
Inventor 张京宏韩飞
Owner 韩飞
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