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Purification method of thymalfasin

A technology of thymus method and purified water, which is applied in the preparation method of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problem of low purity of thymus method products, low yield of thymus method products, and low yield of thymus method. Can not be effectively separated and other problems, to achieve the effect of low impurity, high yield, high purity

Active Publication Date: 2015-05-13
ADLAI NORTYE BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The technical problem that the present invention needs to solve is: select a kind of new purifying method of thymus method, solve (1) the new product purity of thymus method is not high, (2) the yield of new product of thymus method is not high, (3) thymus method is new and The problem that the impurity D-Asn28-thymus fasin cannot be effectively separated

Method used

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  • Purification method of thymalfasin
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  • Purification method of thymalfasin

Examples

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Embodiment 1

[0120] 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 (Thymofasin) Preparation

[0121] step 1

[0122] Synthesis of Fmoc-Asp(RinkAmide-MBHA-Resin)-OtBu

[0123]Weigh Rink Amide MBHA resin (substitution degree 0.45mmol / g) (2.67g, 1.0equiv) into a solid-phase synthesis tube, add DCM (10mL), stir and swell at a temperature of 10-30°C for 30min, and remove the liquid by suction filtration. Then add DMF (10mL×2) to wash. Measure 20% (v / v, the same below) Piperidine / DMF solution (9.0 mL), add it to the solid-phase synthesis tube, control the temperature at 20-30 °C and stir for 5 min, then remove the liquid by suction filtration. Measure 20% Piperidine / DMF solution (9.0 mL) again, add it to the solid-phase synthesis tube, control the temperature at 20-30°C and stir for 10 minutes, then remove the liquid by suction filtration. Measure DMF (10mL×7) into the solid-phase synthesis tube for washing, and remove t...

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Abstract

The invention relates to a new purification method of the thymus method, which is characterized in that it comprises the following steps: step 1, dissolving the thymus method crude product in water, and adjusting the pH to 3-5; step 2, setting a gradient according to the volume fraction, and using mobile phase A to wash the polymer Reverse-phase chromatographic column; step 3, load the solution of step 1 into the polymer reverse-phase chromatographic column; step 4, set the gradient according to the volume fraction, the initial state of the elution gradient mobile phase B is 5%, keep for 2 minutes, and then Increase the proportion of mobile phase B to 25% within 5 minutes, then increase the proportion of mobile phase B to 40% within 30 minutes. Collect the eluted fractions in parts; step 5, dry the eluted fractions to obtain pure thymusfasin.

Description

Technical field: [0001] The invention relates to a purification method of polypeptide drugs, in particular to a new purification method of thymus method. Background technique: [0002] Thymalfasin, British name: Thymalfasin, is a linear polypeptide with the following structural formula: [0003] [0004] Can be abbreviated as: [0005] N-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 [0006] Thymosin α1 (also known as thymosin α1) is the main biologically active component of thymosin and an important immunomodulatory substance in the body. It is a polypeptide composed of 28 amino acids acetylated at the nitrogen end. Studies have shown that thymus fasin promotes the development and differentiation of bone marrow stem cells into pro-lymphocytes and pro-lymphocytes; induces the differentiation and maturation of T lymphocytes, and further differentiates mature T cells into several different subgroups...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/575C07K1/20
Inventor 路杨杨东晖陈晓航
Owner ADLAI NORTYE BIOPHARMA CO LTD
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