Thymalfasin peptide resin pyrolysis method

A thymosin and peptide resin technology, applied in the field of peptide solid-phase synthesis, can solve problems such as hidden dangers of explosion safety, product degradation, and long time, and achieve the effect of improving purity, reducing dosage, and eliminating centrifugation.

Inactive Publication Date: 2019-05-17
SINOPEP ALLSINO BIOPHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ether solutions such as ether MTBE have potential safety hazards of explosion;
[0014] 2. Vacuum drying takes a long time, usually more than 8 hours;
[0015] 3. Due to the long time of the crude peptide state, the product is degraded and a large Asp is produced. 28 - Thymofasin

Method used

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  • Thymalfasin peptide resin pyrolysis method
  • Thymalfasin peptide resin pyrolysis method
  • Thymalfasin peptide resin pyrolysis method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, a kind of cracking method of thymofaxin peptide resin, its steps are as follows:

[0044](1) Take thymofasin resin and add 6 times the volume of lysate, crack at 10°C for 3 hours, filter off the resin to obtain the filtrate; the lysate is composed of TFA and TIS, and the volume ratio TFA:TIS=93:7;

[0045] (2) under stirring, add the filtrate to the ether at -10°C which is 6 times the volume of the filtrate; the ether is diethyl ether;

[0046] (3) under stirring, add 0.5M sodium bicarbonate solution of 0.5 times volume to the above solution;

[0047] (4) under stirring, add alkaline solution until the solution is clear; alkaline solution is sodium bicarbonate solution;

[0048] (5) Turn off the stirring, leave to stand for stratification, take the upper organic phase and add an appropriate amount of water to wash, and the water added in the washing is 30 times the quality of the resin; the lower water phase is combined, and the residual ether is removed ...

Embodiment 2

[0050] Embodiment 2, a kind of cracking method of thymofaxin peptide resin, its steps are as follows:

[0051] (1) Take thymofasin resin and add 12 times the volume of lysate, lyse at 30°C for 1 hour, filter out the resin to obtain the filtrate; the lysate is a mixed type composed of TFA and a combination of EDT, TA, and TC The lysate, wherein the content of TFA accounts for 90% of the mixed lysate;

[0052] (2) under stirring, the filtrate is added in the ether of 10 ℃ of 8 times of filtrate volume; Described ether is MTBE or sherwood oil;

[0053] (3) under stirring, add 1 times the volume of 1.5M ammonia solution to the above solution;

[0054] (4) under stirring, add alkaline solution until the solution is clear; alkaline solution is ammonia solution;

[0055] (5) Turn off the stirring, leave to stand for stratification, take the upper organic phase and add an appropriate amount of water to wash, and the water added in the washing is 40 times the quality of the resin; th...

Embodiment 3

[0057] Embodiment 3, a kind of cracking method of thymofasin resin, its steps are as follows:

[0058] (1) Take thymofasin resin and add 10 times the volume of lysate, crack at 25°C for 2 hours, filter off the resin to obtain the filtrate; the lysate is a mixed lysate composed of TFA and TIS, and the volume ratio TFA:TIS =95:5.

[0059] (2) under stirring, add the filtrate to the ether with 10 times the volume of the filtrate at 0° C.; the ether is selected from diethyl ether, MTBE, petroleum ether or isopropyl ether;

[0060] (3) under stirring, add 0.6 times the volume of 1.0M sodium bicarbonate or ammonia solution to the above solution;

[0061] (4) under stirring, add alkaline solution until the solution is clear; alkaline solution is sodium bicarbonate solution, ammonia solution, sodium hydroxide solution or sodium carbonate solution;

[0062] (5) Turn off the stirring, leave to stand for stratification, take the upper organic phase and add an appropriate amount of wate...

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Abstract

The invention discloses a thymalfasin peptide resin pyrolysis method, and belongs to the field of polypeptide solid-phase synthesis. The method comprises the steps of taking thymalfasin peptide resin,adding a lysate which is 6-12 times the volume of the resin, conducting pyrolysis at 10-30 DEG C for 1-3 h, and filtering out resin to obtain a filtrate; adding the filtrate into ether which accountsfor 6 times or more of the volume of the filtrate; adding a 0.5M-1.5M sodium bicaronate or ammonium hydroxide solution which is 0.5-1 time the volume of the solution into the solution; adding an alkaline solution until the solution is clarified; conducting standing and layering, taking an upper-layer organic phase, adding a moderate amount of water for washing, merging the upper-layer organic phase with a lower-layer aqueous phase, and conducting vacuum pressure reduction to remove residual ether; adding acid to adjust the pH value to 2-8, and conducting filtering and purification. By means of the method, there is no need to use ether to wash crude peptide, the use amount of the ether is greatly reduced, the centrifuging operation is omitted at the same time, and the fire blast risk of the ether is lowered; the crude peptide drying step is omitted, the production labor hour is shortened, the risk of thymalfasin degradation is also lowered, and the purity of crude products is improved.

Description

Technical field: [0001] The invention relates to the field of solid-phase synthesis of polypeptides, in particular to a thymus method new peptide resin cracking method. Background technique: [0002] Thymus Faxin formerly known as thymosin α1, also known as thymus 28 peptide, thymosin α1 or Tα1. Thymus fasin is a kind of thymosin, a natural polypeptide produced by human thymus and present in the thymus circulatory system in the human body. Thymus fasin can promote the transformation of lymphocytes, enhance the phagocytic activity of macrophages, and can be used to treat various immunodeficiency diseases. [0003] In the treatment of chronic hepatitis B with thymus fasin alone, domestic and foreign studies have shown that the sustained response rate is about 37%, which is similar to that of interferon alone, and there is no toxic side effect of interferon. The clinical research on the application of thymusfasin in the treatment of chronic hepatitis B in Taiwan, China shows ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/575C07K1/34
Inventor 王蔡典姜建军玄其存赵呈青
Owner SINOPEP ALLSINO BIOPHARMACEUTICAL CO LTD
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