Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Low anticoagulant heparin and oligosaccharides thereof, and preparation methods and application of low anticoagulant heparin and oligosaccharides thereof in preparation of anti-Alzheimer's disease drugs

A low anticoagulant and oligosaccharide technology, applied in the field of medicine, can solve the problems of low anticoagulant heparin by-products, waste of biological resources, etc., and achieve the effect of abundant raw material sources, low cost, and avoiding bleeding

Active Publication Date: 2016-03-16
OCEAN UNIV OF CHINA
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] my country is the main producer and exporter of crude heparin in the world. In the refining process of crude heparin, there are a large number of by-products of low anticoagulant heparin. Since no new use has been found, most of them are discarded, resulting in waste of biological resources.

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
  • Low anticoagulant heparin and oligosaccharides thereof, and preparation methods and application of low anticoagulant heparin and oligosaccharides thereof in preparation of anti-Alzheimer's disease drugs
  • Low anticoagulant heparin and oligosaccharides thereof, and preparation methods and application of low anticoagulant heparin and oligosaccharides thereof in preparation of anti-Alzheimer's disease drugs
  • Low anticoagulant heparin and oligosaccharides thereof, and preparation methods and application of low anticoagulant heparin and oligosaccharides thereof in preparation of anti-Alzheimer's disease drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Preparation of low anticoagulant heparin

[0035] The preparation method of low anticoagulant heparin of the present invention specifically comprises the following steps:

[0036] 1. Chondroitinase ABC enzyme removes chondroitin impurities: dissolve pig intestinal mucosa crude heparin (10g) purchased from a heparin production plant in 3L enzymolysis buffer (0.05mol L -1 Ammonium acetate, 2mmol·L -1 Calcium chloride), 50 Units of chondroitinase ABC were added thereto, reacted on a shaking table at 35° C. for 6 hours, and inactivated in a boiling water bath for 10 minutes.

[0037] 2. Hydrochloric acid precipitation to remove nucleic acid impurities: centrifuge the enzymatic solution and take the supernatant. Add 1 mol·L dropwise to the supernatant with stirring -1 hydrochloric acid until the pH of the enzymolysis solution is 3.0, centrifuge, take the supernatant and use 1mol·L -1 neutralized to pH 7.0 with NaOH.

[0038] 3. Ethanol precipitation: After co...

Embodiment 2

[0040] Example 2: Structural characterization of the low anticoagulant heparin

[0041](1) Absolute molecular weight determination and purity analysis: The absolute molecular mass of the low anticoagulant heparin was determined and the purity was analyzed by high performance gel permeation chromatography (HPGPC)-octadecanine laser light scattering method (MALLs).

[0042] Chromatographic conditions: Chromatographic column: ShodexOHPakSB804HQ column and ShodexOHPakSB802.5HQ column in series; mobile phase: 0.1mol L -1 Na 2 SO 4 Aqueous solution; The molecular weight of low anticoagulant heparin is detected and determined in series by a differential detector and an octagonal laser light scattering instrument in series; the sample is in the middle of a single symmetrical peak in the HPGPC spectrogram (such as figure 1 Shown), the sample purity is very high, and its absolute molecular mass is measured as 10.24kDa.

[0043] (2) Analysis of disaccharide composition: make low antic...

Embodiment 3

[0049] Example 3: Anticoagulant activity of low anticoagulant heparin

[0050] Follow the steps indicated in the kit to determine the inhibitory effect of the sample on the activity of FXa and FIIa, the initial concentration of the sample is 0.001, 0.01, 0.1, 1, 10 and 100 μg·mL -1 . Add 20 μL of antithrombin III (ATIII) and 20 μL of sample solution to each well of a 96-well plate, incubate at 37°C for 1 min, then add 40 μL of FXa or FIIa, incubate at 37°C for 1 min, add 100 μL of FXa or FIIa substrate, and incubate at 37°C 4min, finally add 100μL stop solution, detect absorbance at 405nm wavelength, calculate inhibition rate: result ( Figure 4 ) showed that LAH had weak anticoagulant activity, which was much lower than that of HP and ES standards, and slightly higher than that of HS standards.

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
Molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to low anticoagulant heparin and oligosaccharides thereof, and preparation methods and an application of the low anticoagulant heparin and the oligosaccharides thereof in preparation of anti-Alzheimer's disease drugs. A porcine small intestinal mucosa crude product heparin is treated by chondroitinase ABC to remove chondroitin sulfate and hyaluronic acid, subjected to hydrochloric acid precipitation to remove nucleic acid, subjected to ethanol precipitation and subjected to anion exchange resin purification, and finally the high-purity low anticoagulant heparin is obtained; furthermore, the low anticoagulant heparin is degraded to obtain the oligosaccharides with different molecular weights by heparinase or a beta-removal method; the obtained low anticoagulant heparin has the molecular weight of 10-20 kDa, the oligosaccharides have the molecular weight of 1-7 kDa, and each disaccharide unit averagely contains 1.5-2.5 sulfates. The prepared low anticoagulant heparin and the oligosaccharides thereof can significantly inhibit the BACE1 activity through experimental evidence and can be used for preparation of the anti-Alzheimer's disease drugs.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a low-anticoagulant heparin, its oligosaccharides, a preparation method thereof, and an application in preparation of anti-Alzheimer's medicine. Background technique [0002] Heparin (Heparin, HP) has a variety of biological functions. Currently, it is mainly used as an anticoagulant to prevent thrombosis. In addition, it also has various functions such as adjusting blood lipid concentration, enhancing immunity, anti-inflammation, and anti-allergy. However, due to side effects such as hemorrhage, thrombocytopenia, hyperkalemia and osteoporosis in the clinical application of heparin, its clinical application in other active aspects is limited. Therefore, research on low anticoagulant heparin (Lowanticoagulantheparin, LAH) products has become a hot spot. Compared with heparin, low anticoagulant heparin has the advantages of lower anticoagulant activity, does not bind ...

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): C08B37/10A61K31/727A61P25/28
Inventor 于广利张晓李国云蔡超李芹英
Owner OCEAN UNIV OF CHINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com