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Purification method for atosiban

A purification method and solution technology, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as failure to meet industrialization requirements, low peptide purity, and low application value, and achieve simple and feasible operation, The effect of high purity

Active Publication Date: 2012-07-18
HYBIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the purity of the atosiban target peptide obtained by the above method is low, which cannot meet the requirements of industrialization, and the application value is not high

Method used

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  • Purification method for atosiban
  • Purification method for atosiban

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Dissolve the crude peptide at a concentration of 100 g / L with 50% acetic acid aqueous solution by volume, stir to dissolve the sample completely, filter it with a filter membrane, and collect the filtrate. Add ferric chloride reagent according to the proportion of 5% of the weight of the crude peptide, and dilute the volume ratio of the acetic acid in the crude peptide solution to 40% with water for later use.

[0037] A chromatographic column with octadecylsilane bonded silica gel as the stationary phase, the chromatographic column is: 5cm×25cm (diameter×length). Rinse the chromatographic column with more than 50% acetonitrile, and then equilibrate the sample, and the sample load is 1.5-3g. The mixed solution with perchlorate aqueous solution and acetonitrile is mobile phase, and wherein mobile phase A phase is 0.4% perchloric acid aqueous solution (v / v), adjusts pH value 2.5 with the sodium hydroxide aqueous solution of 50mmol; Mobile phase B phase is Acetonitrile. Th...

Embodiment 2

[0040] Dissolve the crude peptide at a concentration of 100 g / L with 80% acetic acid aqueous solution by volume, stir to dissolve the sample completely, filter it with a filter membrane, and collect the filtrate. The ferric sulfate reagent is added according to the ratio of 4% of the weight of the crude peptide, and the volume ratio of the acetic acid in the crude peptide solution is diluted to 30% by water for use.

[0041] A chromatographic column with octaalkylsilane bonded silica gel as the stationary phase, the chromatographic column is: 15cm×25cm (diameter×length). Rinse the chromatographic column with more than 50% acetonitrile, and then balance the sample, and the sample load is 25-40g. With the mixed solution of perchlorate aqueous solution and acetonitrile as mobile phase, wherein mobile phase A phase is 0.3% perchloric acid aqueous solution (v / v), adjust pH value 3.2 with the potassium hydroxide aqueous solution of 50mmol; Mobile phase B phase is Acetonitrile. The...

Embodiment 3

[0044] Dissolve the crude peptide at a concentration of 100 g / L with 90% acetic acid aqueous solution by volume, stir to dissolve the sample completely, filter it with a filter membrane, and collect the filtrate. Add ferric chloride reagent according to the ratio of 8% of the weight of the crude peptide, and dilute the volume ratio of the acetic acid in the crude peptide solution to 40% with water for later use.

[0045] A chromatographic column with octadecylsilane bonded silica gel as the stationary phase, the chromatographic column is: 30cm×25cm (diameter×length). Rinse the chromatographic column with more than 50% acetonitrile, and then equilibrate the sample, and the sample load is 80-170g. With the mixed solution of perchlorate aqueous solution and acetonitrile as mobile phase, wherein mobile phase A phase is 0.8% perchloric acid aqueous solution (v / v), adjust pH value 3.0 with the potassium hydroxide aqueous solution of 50mmol; Mobile phase B phase is Acetonitrile. Th...

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Abstract

The invention belongs to the technical field of pharmaceutical chemistry, and discloses a purification method for atosiban. The purification method comprises the following steps of: treating atosiban coarse peptide solution by taking trivalent iron ion salt as a reducing agent; removing impurities which are not fully oxidized in the atosiban coarse peptide solution; purifying the atosiban coarse peptide solution by using high performance liquid chromatography; and converting atosiban perchlorate prepared through purification to prepare an atosiban pure product by using the high performance liquid chromatography. The purification method provided by the invention is easy and feasible to operate; the prepared atosiban pure peptide product has high purity; meanwhile, the yield of the atosiban pure peptide product prepared by the method is high; and over 1,000 g of the atosiban pure peptide product can be obtained in one batch and is sufficient to meet the industrial requirements.

Description

technical field [0001] The invention belongs to the technical field of medicinal chemistry, and in particular relates to a purification method of atosiban. Background technique [0002] Atosiban, English name Atosiban, chemical name: 1-(3-mercaptopropanol acid)-2-(0-ethyl-D-tyrosine)-4-L-threonine-8-L - Ornithine - Oxytocin. Atosiban is a synthetic cyclic polypeptide. The peptide chain contains three unnatural amino acids D-Tyr (Et), Mpa and Orn and a pair of disulfide bonds forming a ring between Mpa and Cys. The molecular formula is C 43 h 67 N 11 o 12 S 2 , the molecular weight is 994.19, and its structural formula is as follows: [0003] [0004] Atosiban is a breakthrough product in obstetrics and gynecology. It is an oxytocin analogue, and it is an oxytocin competitive antagonist of receptors on the decidua and fetal membranes in the uterus. It can directly interact with oxytocin Competing for oxytocin receptors, inhibiting the combination of oxytocin and oxy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/16C07K1/36C07K1/16C07K1/20
CPCC07K7/16C07K7/06C07K1/20
Inventor 赵忠卫刘建马亚平袁建成
Owner HYBIO PHARMA
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