Method for preparing high-purity thymalfasin

A new and high-purity technology of thymus method, which is applied in the preparation method of peptides, thymosin, chemical instruments and methods, etc., can solve the problems such as the difficulty of synthesizing peptides

Active Publication Date: 2014-06-25
郑州大明药物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] At present, there are mainly solid-phase synthesis methods for chemically synthesizing thymosin. Although this method is relatively mature, it is very difficult to use this solid-phase synthesis method to synthesize polypeptides.

Method used

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  • Method for preparing high-purity thymalfasin
  • Method for preparing high-purity thymalfasin
  • Method for preparing high-purity thymalfasin

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Embodiment

[0076] The present invention prepares the new method of high-purity thymus method, and the detailed steps of this preparation method are as follows:

[0077] (1) Preparation of polypeptide fragment-Asn-Glu-Ala-Glu-Glu-Val-Val-Glu-Lys-Lys-Glu resin:

[0078] a. First add Fmoc-Asn(Trt)-wang resin resin into the container, then add dimethylformamide to swell, the swelling time is 30min; the ratio of the amount of the resin to dimethylformamide For 1g resin: 10mL dimethylformamide;

[0079]b, the resin after step a swelling is added in the mixed solution of piperidine and dimethylformamide to carry out deprotection, the mass ratio between piperidine and dimethylformamide in the mixed solution is 1:1, every gram of Fmoc -Asn(Trt)-wang resin was added to 10mL of the mixture, and after deprotection, it was washed 5 times with dimethylformamide;

[0080] c. Add the deprotected and washed resin in step b into the reactor, then add Fmoc-Glu(OtBu)-OH amino acid, condensation agent HATU...

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Abstract

The invention discloses a method for preparing high-purity thymalfasin. The preparation method comprises: firstly preparing a first resin polypeptide fragment, a second resin polypeptide fragment, a third resin polypeptide fragment, a fourth resin polypeptide fragment and a fifth resin polypeptide fragment respectively; cleaving the second resin polypeptide fragment, the third resin polypeptide fragment, the fourth resin polypeptide fragment and the fifth resin polypeptide fragment to obtain crude peptide fragments; purifying the obtained second, third, fourth and fifth crude polypeptide fragments respectively; linking each purified polypeptide fragment to the first resin polypeptide fragment; carrying out an acetylation reaction on resin polypeptide fragments to obtain thymalfasin wang resin; cleaving to obtain the thymalfasin crude product; then purifying the obtained thymalfasin crude product twice; and collecting the mobile phase containing thymalfasin in a collection and purification process, evaporating to dryness under reduced pressure and centrifuging to obtain thymalfasin, and vacuum drying to obtain thymalfasin products. The thymalfasin is prepared by the method disclosed by the invention, the reaction time is effectively shortened, the reaction yield and the quality of the final product are improved; and the purity of thymalfasin prepared is more than 99%.

Description

technical field [0001] The invention relates to the technical field of polypeptide solid-phase synthesis, in particular to a new method for preparing high-purity thymus. Background technique [0002] Thymosin is a group of physiologically active polypeptides secreted by thymus tissue, which can promote lymphocyte transformation, enhance macrophage phagocytosis, and can be used to treat various immunodeficiency diseases. [0003] The new chemical structural formula of thymus method is: [0004] Ac-Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu-Lys-Glu-Lys-Lys-Glu-Val-Val-Glu- Glu-Ala-Glu-Asn-OH. [0005] In the treatment of chronic hepatitis B with thymusfasin alone, domestic and foreign studies have shown that the sustained response rate of thymusfasin alone in the treatment of chronic hepatitis B is about 37%, which is similar to that of interferon alone, but without the toxic side effects of interferon. [0006] The clinical research on the application ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/575C07K1/06C07K1/04
CPCC07K14/57581
Inventor 张宝国李沁沁王艳侨翟腾朱赞梅张亚惠苗海敏王娜
Owner 郑州大明药物科技有限公司
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