Preparation and purification method of heparin hexasaccharides containing N-unsubstituted glucosamine

A technology of glucosamine heparin and heparin pyridine hexasaccharide, which is applied in the field of preparation and purification of natural products, and can solve problems such as high price

Inactive Publication Date: 2014-04-16
FUZHOU UNIV
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, the methods for preparing heparin oligosaccharides are many steps and expensive
There are few reports on direct...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation and purification method of heparin hexasaccharides containing N-unsubstituted glucosamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] To prepare GlcNH containing 3 + Heparin hexasaccharide as an example:

[0041] 1 Preparation and purification of heparin sodium hexasaccharide

[0042] Heparinase Ⅰ, Ⅱ, Ⅲ Enzymolysis of low molecular weight heparin (3-8kD) at 37°C for 48h (commercially available) enzymatic hydrolysis of low molecular weight heparin through gel chromatography column (Bio-Gel P-10), the mobile phase is 0.2M NH 4 HCO 3 , at UV 232nm, measure the absorbance of the eluate. Collect the elution peak of heparin sodium hexasaccharide mixture, and then remove NH in 55°C water bath for 24h×3 4 HCO 3 , Freeze-dried at -80°C for 5 h. Then use SAX-HPLC, two-phase linear gradient elution under the conditions of 0-0.6M NaCl, 0.6-1.3M NaCl, separate and collect the highly sulfated heparin sodium hexasaccharide crude product. The specific parameters are as follows:

[0043] Chromatographic column: ProPac PA1 (9×250mm) Semi-Prep

[0044] Flow rate: 4ml / min

[0045] Mobile phase: A:pH3.5H ...

Embodiment 2

[0075] To prepare GlcNH containing 3 + Heparin hexasaccharide as an example:

[0076] 1 Preparation and purification of heparin sodium hexasaccharide

[0077] Heparinase Ⅰ, Ⅱ, Ⅲ Enzymolysis of low molecular weight heparin (3-8kD) at 37°C for 48h (commercially available) enzymatic hydrolysis of low molecular weight heparin through gel chromatography column (Bio-Gel P-10), the mobile phase is 0.2M NH 4 HCO 3 , at UV 232nm, measure the absorbance of the eluate. Collect the elution peak of heparin sodium hexasaccharide mixture, and then remove NH in 55°C water bath for 24h×3 4 HCO 3 , Freeze-dried at -80°C for 5 h. Then use SAX-HPLC, two-phase linear gradient elution under the conditions of 0-0.6M NaCl, 0.6-1.3M NaCl, separate and collect the highly sulfated heparin sodium hexasaccharide crude product. The specific parameters are as follows:

[0078] Chromatographic column: ProPac PA1 (9×250mm) Semi-Prep

[0079] Flow rate: 4ml / min

[0080] Mobile phase: A: pH3.5 H...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Wavelengthaaaaaaaaaa
Login to view more

Abstract

The invention relates to a preparation and purification method of heparin hexasaccharides containing (i)N(i)-unsubstituted glucosamine (GlcNH3+). The preparation and purification method comprises the following steps: using enzymolysis low molecular weight heparin as the raw material, separating and purifying the heparin sodium hexasaccharide by methods of gel chromatography and ion exchange HPLC; preparing heparin pyridine hexasaccharide by using cation exchange resin and pyridine neutralization process; finally preparing heparin hexasaccharides with different numbers of GlcNH3+ by the (i)N(/i)-sulfate removal method, and separating and purifying the heparin hexasaccharide by the SAX-HPLC method. The preparation and purification method can specifically prepare the heparin oligosaccharides containing different numbers and sequences of GlcNH3+, and provide important oligosaccharides libraries for researching the structure and function of the GlcNH3+ residues in the heparin sulfate and the relationship between the GlcNH3+ residues and diseases.

Description

technical field [0001] The present invention relates to the preparation of GlcNH-containing 3 + The method for heparin oligosaccharide and its purification belong to the field of preparation and purification of natural products. Background technique [0002] There are 12 representative disaccharides in heparan sulfate, including GlcNAc, GlcNSO 3 The disaccharide of the residue is the main form, while the positively charged GlcNH 3 + The disaccharide content of the residue is minimal and its presence varies depending on the cell and tissue of origin. Although GlcNH 3 + The content of heparan sulfate in heparan sulfate is very small, but it is closely related to the invasion of viruses, the occurrence of inflammation, brain tissue damage and other diseases, and plays an important role in biology and pathophysiology in organisms. GlcNH 3 + The research on the structure and function of carbohydrate compounds is of great significance in the treatment of diseases and the ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C08B37/10
Inventor 魏峥梁群焘林江慧魏可镁
Owner FUZHOU UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap