Ularitide preparation method

A technology of uralitide and purification method, which is applied in the field of biomedicine, can solve the problems of low purity and long time, and achieve the effect of simplified process technology and easy control

Inactive Publication Date: 2015-02-25
JIANGSU HANBON SCI & TECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a new method for purifying high-purity ularitide by using a liquid chromatography system to solve the problems of long time and low purity in the process of purifying polypeptide substances such as ularitide. The method includes the following steps:

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] 1. Take 1g of crude ralitide and dissolve it in water to prepare a saturated solution, put it in a filter device, and filter to remove solid particles;

[0010] 2. Pump the ularitide solution into the dynamic axial compression column to prepare the chromatographic system. The column packing size is Φ50×250mm, the filler is octadecyl silica gel, the particle size is 10um, and the sample volume is 1g. The volume ratio of the mobile phase was 10:90~30:70 for gradient elution for 80 minutes. After ulinaritide was eluted, the mobile phase was switched to acetonitrile with a high proportion to wash out the rear-end impurities, and a separation cycle ended. The detection wavelength of the ultraviolet-visible photometric detector used is 215nm, and the fractions with a retention time of 37-45min are collected, and the purity is ≥98.7% by HPLC analysis.

Embodiment 2

[0012] 1. Take 1.5g of crude ralitide and dissolve it in water to prepare a saturated solution, place it in a filter device, and filter to remove solid particles;

[0013] 2. Pump the ularitide solution into the dynamic axial compression column to prepare the chromatographic system. The column packing size is Φ50×250mm, the packing is octadecyl silica gel, the particle size is 10um, and the sample loading is 1.5g. The volume ratio of the mobile phase was 5:95-30:70 for gradient elution for 80 minutes. After ulinaritide was eluted, the mobile phase was switched to acetonitrile with a high proportion to wash out the rear-end impurities, and a separation cycle ended. The detection wavelength of the ultraviolet-visible photometric detector used was 215nm, and the fractions with a retention time of 42-50min were collected, and the purity was ≥98.5% by HPLC analysis.

Embodiment 3

[0015] 1. Dissolve 2.5g of crude ralitide in water, prepare a saturated solution, put it in a filter device, and filter to remove solid particles;

[0016] 2. Pump the ularitide solution into the dynamic axial compression column to prepare the chromatographic system. The column packing size is Φ150×250mm, the packing is octadecyl silica gel, the particle size is 10um, and the sample loading is 2.5g. The volume ratio of the mobile phase was 5:95-30:70 for gradient elution for 100 minutes. After ulinaritide was eluted, the mobile phase was switched to acetonitrile with a high proportion to wash out the rear-end impurities, and a separation cycle ended. The detection wavelength of the ultraviolet-visible photometric detector used was 215nm, and the fractions with a retention time of 51-59min were collected, and the purity was ≥98.2% by HPLC analysis.

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Abstract

The invention discloses a purifying method of ularitide, appropriate column separation conditions can be obtained by optimization of conditions, and the appropriate column separation conditions are further enlarged onto a dynamic axial compression industrial preparative chromatography system for separation for obtaining of high purity ularitide. The advantages of the method are that, good effect of separation and purification can be achieved by direct use of the dynamic axial compression industrial preparative chromatography for separation of an extracted crude product, the method has the advantages of simple operation, short period and high efficiency, and the purity can reach more than 98%.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for purifying ularitide. technical background [0002] Uralitide is a natriuretic cyclic peptide composed of 32 amino acid residues, which was originally isolated from urine by Schulz-Knappe et al. in 1988 and belongs to the atrial natriuretic peptide (ANP) family. Renal natriuretic peptide. Endogenous uraritide is synthesized in renal distal tubular cells, secreted after the lumen, and binds to the downstream natriuretic peptide A receptor in the inner medullary collecting duct, which can regulate the excretion of sodium and water in the kidney. Therefore, uraritide It has vasodilation and natriuretic effects, and it has been confirmed that ularitide can reduce the reabsorption of urine by the kidneys. The clinical treatment of decompensated heart failure (DHF) is aimed at relieving symptoms and stabilizing the patient's hemodynamics. The currently use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/58C07K1/16
CPCC07K14/58
Inventor 张宇刘根水祁威
Owner JIANGSU HANBON SCI & TECH CO
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