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Method for preparing Thymalfasin

A thymus method and resin technology, applied in the field of peptide chemistry, can solve the problems of high cost, cumbersome preparation process of thymus method and low efficiency.

Pending Publication Date: 2021-08-31
JIANGSU GENSCRIPT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a simple and efficient new preparation method of the thymus method to solve the problems of cumbersome, low efficiency or high cost of the current new preparation method of the thymus method

Method used

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  • Method for preparing Thymalfasin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1: preparation of thymus fasin

[0069] (1) Preparation of Fmoc-Asp-resin complex

[0070] Add 1 g of Rink amide AM resin (0.5 mmol / g) into a reactor with a filter device, inject 10 ml of DMF, let stand for 1 h, and filter.

[0071] Add 10ml of PIP / DMF solution with a volume ratio of 1:4 into the reactor, and react at 30°C for 20min. After the reaction is complete, filter. Then add 10ml DMF to wash the resin six times and filter.

[0072] Take 4mmol Fmoc-Asp-OtBu, 4mmol HOBt and 4mmol DIC, dissolve in 10ml DMF, add to the resin, and react at 30°C for 2h. After the reaction, filter. Then add 10ml of DMF, wash the resin six times, and filter.

[0073] (2) Preparation of protected thymus fasin-resin complex

[0074] Add 10ml of PIP / DMF solution with a volume ratio of 1:4 into the reactor, and react at 30°C for 20min. After the reaction is complete, filter. Then add 10ml DMF to wash the resin six times and filter.

[0075] Take 4mmol Fmoc-Glu(OtBu)-OH, 4mm...

Embodiment 2

[0081] Embodiment 2: preparation of thymus fasin

[0082] (1) Preparation of Fmoc-Asp-resin complex

[0083] Add 1 g of Rink amide MBHA resin (0.5 mmol / g) into a reactor with a filter device, inject 10 ml of DMF, let stand for 1 h, and filter.

[0084] Add 10ml of PIP / DMF solution with a volume ratio of 1:3 into the reactor, and react at 20°C for 10min. After the reaction is complete, filter. Then add 10ml DMF to wash the resin six times and filter.

[0085] Take 3mmol Fmoc-Asp-OtBu, 3mmol HOBt and 3mmol DIC, dissolve in 10ml DMF, add to the resin, and react at 20°C for 4h. After the reaction, filter. Then add 10ml of DMF, wash the resin six times, and filter.

[0086] (2) Preparation of protected thymus fasin-resin complex

[0087] Add 10ml of PIP / DMF solution with a volume ratio of 1:3 into the reactor, and react at 20°C for 10min. After the reaction is complete, filter. Then add 10ml DMF to wash the resin six times and filter.

[0088] Take 3mmol Fmoc-Glu(OtBu)-OH,...

Embodiment 3

[0094] Embodiment 3: preparation of thymus fasin

[0095] (1) Preparation of Fmoc-Asp-resin complex

[0096] Add 1 g of amino 2-chlorotrityl resin (0.5 mmol / g) into a reactor with a filter device, inject 10 ml of DMF, let stand for 1 h, and filter.

[0097] Add 10ml of PIP / DMF solution with a volume ratio of 1:1 into the reactor, and react at 60°C for 60min. After the reaction is complete, filter. Then add 10ml DMF to wash the resin six times and filter.

[0098] Take 6mmol Fmoc-Asp-OtBu, 6mmol HBTU and 12mmol DIEA, dissolve in 10ml DMF, add to the resin, and react at 60°C for 1h. After the reaction, filter. Then add 10ml of DMF, wash the resin six times, and filter.

[0099] (2) Preparation of protected thymus fasin-resin complex

[0100] Add 10ml of PIP / DMF solution with a volume ratio of 1:1 into the reactor, and react at 60°C for 60min. After the reaction is complete, filter. Then add 10ml DMF to wash the resin six times and filter.

[0101] Take 6mmol Fmoc-Glu(Ot...

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Abstract

The invention discloses a method for preparing Thymalfasin. The method comprises the following steps: (1) preparing a Fmoc-Asp-resin complex; (2) preparing a protected Thymalfasin-resin complex; (3) removing a lateral-chain protective group by employing a cutting reagent, splitting Thymalfasin from resin, and carrying out ethyl ether precipitation, so as to obtain a Thymalfasin crude product; and (4) purifying the crude product by employing high-performance liquid chromatography, thereby obtaining refined Thymalfasin. The method for preparing the Thymalfasin, provided by the invention, is simple, convenient and efficient, and the problem that Thymalfasin preparation processes are tedious and are low in efficiency and high in cost at present is solved.

Description

technical field [0001] The invention belongs to the field of polypeptide chemistry, and in particular relates to a new preparation method of thymus method. Background technique [0002] Thymofasin consists of 28 amino acids, the N-terminal contains acetylation structure, the molecular weight is 3108, and the primary structure is Ac-Ser 1 -Asp 2 -Ala 3 -Ala 4 -Val 5 -Asp 6 -Thr 7 -Ser 8 -Ser 9 -Glu 10 -Ile 11 -Thr 12 -Thr 13 -Lys 14 -Asp 15 -Leu 16 -Lys 17 -Glu 18 -Lys 19 -Lys 20 -Glu 21 -Val 22 -Val 23 -Glu 24 -Glu 25 -Ala 26 -Glu 27 -Asn 28 -OH. Thymalfasin is a regulator of biological responses, an immune enhancer for T lymphocytes, it can promote the maturation and differentiation of T cells, and can promote mature T cells and NK cells to secrete a variety of lymphokines (IL -2, INF-α, INF-γ, etc.), but also promote the production of high-affinity interleukin-2 receptor (IL-ZR). In addition, thymofasin has a direct inhibitory or killing effect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/575C07K1/04C07K1/06C07K1/20C07K1/14
CPCC07K14/57581Y02P20/55
Inventor 王卫国张藤王翔
Owner JIANGSU GENSCRIPT BIOTECH CO LTD
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