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Microwave-assisted continuous flow large-scale polypeptide synthesis method and device

A synthesis device and microwave-assisted technology, which is applied in the field of microwave-assisted continuous flow large-scale peptide synthesis method and its device, can solve the problems of low output, low efficiency and low yield, etc., and achieve increased output, shortened reaction time, and significant social impact. Effects on Value and Economic Benefits

Inactive Publication Date: 2009-09-23
CHINESE PEPTIDE CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, at present, the output of domestic polypeptide synthesis enterprises is relatively low, and most of them only reach the gram level, while many foreign enterprises have already reached the kilogram level or even tens of kilograms level, but the synthesis efficiency and yield are not very high.

Method used

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  • Microwave-assisted continuous flow large-scale polypeptide synthesis method and device

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation of embodiment 1 Thymopentin

[0041] ① Put 5.25kg of Fmoc-Tyr(tBu)-Wang resin with a degree of substitution of 0.4mmol / g into the reactor, add 30L of deprotecting group reagent, start the pump to circulate the reactants between the microwave reaction center and the reactor, and turn on the microwave The microwave generating device in the reaction center enables it to generate microwaves, and the microwave settings are: 70°C, 1.2MPa. When the reactant passes through the microwave reaction center, the reaction of removing the amino protecting group is accelerated under microwave radiation. After 5 minutes, turn off the pump and stop the microwave radiation, filter off the deprotecting group reagent, add reagent DMF 10.5L to wash the resin 3 times, and drain with nitrogen gas. According to the polypeptide sequence, add pre-activated Fmoc-Val-OH and coupling reagent HBTU to the reaction kettle, the molar ratio of the reagents used is raw resin: Fmoc-Val-OH:...

Embodiment 2

[0044] The preparation of embodiment 2 Thymopentin

[0045] The polypeptide to be synthesized in this example and the operation are exactly the same as in Example 1, only the process conditions used are different.

[0046] ① The time of microwave irradiation is 20 minutes, the resin is washed 5 times after filtering off the deprotecting group reagent, and the resin is washed 3 times after filtering off the residual amino acid and reagent liquid, and the rest are the same as in Example 1.

[0047] 2., 3. steps are with embodiment 1. Obtain 1.38kg of crude polypeptide. The purity measured by liquid chromatography was 92.5%, and the yield was 96%.

Embodiment 3

[0048] Example 3 Preparation of Glucagon-Like Peptide-1 (Glucagon-Like Peptide I)

[0049] ① Put 1.26kg of Fmoc-Arg(Pbf)-Amide resin with a degree of substitution of 0.4mmol / g into the reactor, add 10L of deprotecting group reagent, start the pump to circulate the reactant between the microwave reaction center and the reactor, and turn on the microwave The microwave generating device in the reaction center enables it to generate microwaves, and the microwave settings are: 70°C, 1.2MPa. When the reactant passes through the microwave reaction center, the reaction of removing the amino protecting group is accelerated under microwave radiation. After 1 minute, turn off the pump and stop the microwave radiation, filter off the deprotecting group reagent, add reagent DMF 12.6L to wash the resin 3 times, and drain with nitrogen gas. According to the polypeptide sequence, add pre-activated Fmoc-Gly-OH and coupling reagent HBTU to the reaction kettle, the molar ratio of the reagents i...

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Abstract

The invention provides a microwave-assisted continuous flow large-scale polypeptide synthesis method and a device thereof. The device mainly comprises a reaction kettle, a microwave reaction center and a connecting pipeline, and an on-line detection device and a power device are arranged on the connecting pipeline between the reaction kettle and the microwave reaction center. The invention leads reaction substances to flow cyclically between the reaction kettle and the microwave reaction center, thereby achieving the purposes of locally adding microwave radiation in a reaction system, shortening the reaction time, realizing the large-scale rapid synthesis of target products and increasing the yield of polypeptides to kilograms or tens of kilograms.

Description

technical field [0001] The invention relates to a polypeptide synthesis method and a device thereof, in particular to a microwave-assisted continuous flow large-scale polypeptide synthesis method and a device thereof, belonging to the fields of organic chemistry and medicinal chemistry. Background technique [0002] Polypeptides are biologically active substances involved in various cell functions in organisms. The artificial synthesis of polypeptides has a history of hundreds of years. With the development of biotechnology, peptides have been widely used in the field of medicine, such as peptide vaccines, anti-tumor peptides, anti-viral peptides, peptide-oriented drugs, cytokine mimetic peptides, anti-active peptides, etc. Peptides are not only a target for new drug development, but also a screening target for the development of other drugs. Therefore, the synthesis of peptides has become an important subject of research by scholars from all over the world. [0003] Polyp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/06C07K1/04
Inventor 李湘徐琪卜远波
Owner CHINESE PEPTIDE CO
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