A kind of refining method of vasopressin

A refining method, vasopressin technology, applied in the preparation method of peptide, oxytocin/vasopressin, separation method, etc., can solve the problem of large amount of organic hazardous waste liquid, high cost of waste liquid treatment, uneconomical, etc. question

Active Publication Date: 2021-05-14
SPH NO 1 BIOCHEM & PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome that when refining vasopressin in the prior art, the removal rate of impurities in the crude vasopressin solution is low, and the amount of organic hazardous waste liquid produced is large, resulting in high cost of waste liquid treatment. , uneconomical defects, but provides a refined method of vasopressin

Method used

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  • A kind of refining method of vasopressin
  • A kind of refining method of vasopressin
  • A kind of refining method of vasopressin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Packing of L&L4002 preparative column with inner diameter of 50mm

[0049] Using Load&Lock dynamic axial compression and static locking technology, packing ODS-AQ, pore size 10nm, particle size 10μm, packed to column bed pressure 1000psi, using Varian chromatography packing system, 300g dry powder filler, 600ml isopropanol after stirring and homogenizing, poured into L&L4002 preparative column with inner diameter 50mm, compression ratio 1.5: 1, the carrier gas is N 2 , adjust the carrier gas pressure so that the oil pressure gauge pressure is 1500psi, and the dynamic axial compression to the height of the column bed is 25cm, which is used as a preparative column for reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification schemes.

Embodiment 2

[0050] Example 2 Reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification of crude vasopressin solution

[0051] Instrument: Varian SD-1 high pressure liquid phase preparation system

[0052] Chromatographic column: self-packed preparative column Load&Lock4002 50×250mm, ODS-AQ 10μm 10nm

[0053] The structural formula of vasopressin is The concentration of the crude reduced vasopressin in the crude reduced vasopressin solution is 1.0 mg / ml, and the solvent in the crude vasopressin solution is an aqueous solution containing trifluoroacetic acid and acetic acid.

[0054] The mobile phase A is 0.02% acetic acid / water by volume, and the mobile phase B is 0.02% acetic acid / acetonitrile by volume. Sample C1 is a crude product solution of vasopressin. The HPLC purity determined by the HPLC method is 73.63%, and the mobile phase C2 is 10 mM NH 4 Ac-NH 4 OH pH7.5 aqueous solution.

[0055] The reverse-phase enrichment, reverse-phase salt transfer a...

Embodiment 3

[0059] Example 3 Reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification of crude vasopressin solution

[0060] Instrument: Varian SD-1 high pressure liquid phase preparation system

[0061] Chromatographic column: self-packed preparative column Load&Lock4002 50×250mm, ODS-AQ 10μm 10nm

[0062] The structural formula of vasopressin is The concentration of the crude reduced vasopressin in the crude reduced vasopressin solution is 1.5 mg / ml, and the solvent in the crude vasopressin solution is an aqueous solution containing trifluoroacetic acid and acetic acid.

[0063] Mobile phase A is 0.05% acetic acid / water by volume, and mobile phase B is 0.05% acetic acid / acetonitrile by volume. Sample C1 is a crude vasopressin solution. The HPLC purity determined by the HPLC method is 75.23%, and the mobile phase C2 is 20mM NH 4 Ac-NH 4 OH pH8.5 aqueous solution.

[0064] The reverse-phase enrichment, reverse-phase salt transfer and reverse-phase pur...

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Abstract

The invention discloses a refining method of vasopressin. The refining method comprises the following steps: using high-performance liquid phase chromatography to sequentially carry out reverse-phase enrichment, reverse-phase salt conversion, and reverse-phase purification of the crude vasopressin solution , that is, the crude product solution of vasopressin is obtained by oxidizing the crude product solution of reduced vasopressin synthesized in solid phase; the filler for high performance liquid phase chromatography is a super water-resistant filler. The present invention uses the one-step method of on-line reverse-phase enrichment, reverse-phase salt conversion, and reverse-phase purification to obtain pure polypeptide products. The removal rate of impurities in crude vasopressin products is high, and the purity of pure vasopressin products is higher. The production process, in which the mobile phase in the stages of column balance, sample enrichment and salt conversion is an aqueous solution, which is environmentally friendly and pollution-free, and the effluent waste liquid can be directly treated for sewage and recycled.

Description

technical field [0001] The invention relates to a refining method of vasopressin. Background technique [0002] Vasopressin is a synthetic polypeptide consisting of nine amino acid residues with a chemical structure of The theoretical molecular weight of vasopressin is 1084.24. Belongs to neurohypophyseal hormone, also known as vasopressin or vasopressin, it has two receptors V1 and V2. V1 is mainly distributed on the membrane of vascular smooth muscle cells, and acts through the receptor-G protein-second messenger pathway to constrict blood vessels and increase blood pressure. V2 is found in the epithelial cells of the distal tubule and collecting duct of the kidney, and physiological doses can promote the reabsorption of water by the distal tubule and collecting duct of the kidney, thereby exerting an antidiuretic effect. [0003] At present, most of the common purification methods for peptide drugs on the market use preparative high-performance liquid chromatography, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/16C07K1/20
CPCC07K7/16B01D15/206B01D15/327B01D15/426C07K1/20C07K7/54
Inventor 江锡铭朱鑫磊黄龙丁金国黄臻辉
Owner SPH NO 1 BIOCHEM & PHARMA CO LTD
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