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Low molecular weight imitated stichopus japonicus selenka glycosaminoglycans, preparation method, and applications thereof

A sea cucumber glycosaminoglycan and low-molecular-weight technology, which is applied to medical preparations containing active ingredients, pharmaceutical formulas, organic active ingredients, etc., can solve problems such as the pH is not effectively controlled, the molecular weight of the product is single, and the structure is not clear. , to achieve the effect of prolonging the thromboplastin time and thrombin time, high product purity, and simple and convenient method

Active Publication Date: 2015-05-13
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent CN103285031A established a 5% H 2 o 2 2% acetic acid solution, 60 ° C for 20 hours of free radical degradation system to prepare low molecular weight sea cucumber glycosaminoglycans, and obtain oligosaccharides with a molecular weight of 26000Da~45000Da; and the patent CN101057859A uses 5% H 2 o 2 Low molecular weight sea cucumber glycosaminoglycans with a molecular weight of 8,000-12,000 were prepared by degrading 2% acetic acid solution at 45°C. However, the pH of the two reaction conditions was not effectively controlled, which easily caused the acid degradation of the polysaccharide. The structure is damaged, and the obtained product has a single molecular weight and an unclear structure, and there are certain defects

Method used

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  • Low molecular weight imitated stichopus japonicus selenka glycosaminoglycans, preparation method, and applications thereof
  • Low molecular weight imitated stichopus japonicus selenka glycosaminoglycans, preparation method, and applications thereof

Examples

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Embodiment 1

[0023] Example 1: Preparation of low molecular weight imitation sea cucumber glycosaminoglycan

[0024] Dissolve 10mg of japonicus glycosaminoglycan (HG, weight average molecular weight about 98kDa) in 1mL of 100mmol / L K containing 50mmol / L hydrogen peroxide 2 HPO 4 (pH 7.5) solution, add copper acetate to the aqueous solution to a concentration of 1mmol / L, ultrasonically dissolve, then react at 55°C for 1.5h, add 1mL 20mmol / L EDTA×2Na to terminate the reaction, and centrifuge at 4000rpm for 20 minutes to remove the precipitate . Add 4 times the volume of 95% ethanol to the above supernatant, let stand overnight at 4°C to precipitate, and collect the precipitate by centrifugation. The precipitate was dissolved in a small amount of deionized water, purified by Sephdex G25, concentrated and freeze-dried to obtain low molecular weight imitation sea cucumber glycosaminoglycans DHG-1, DHG-2, DHG-3, and DHG-4. The general structural formula of the low molecular weight imitation ...

Embodiment 2

[0027] Implementation example 2: In vitro anticoagulant activity experiment of low molecular weight imitation sea cucumber glycosaminoglycan

[0028] The main pharmacological activity experimental results of the low-molecular-weight japonicus japonicus glycosaminoglycan prepared by the method of the present invention are as follows, and the following results are all obtained through systematic experimental research according to the method stipulated by the state.

[0029] A. In vitro anticoagulation experiment of low molecular weight imitation sea cucumber glycosaminoglycan

[0030]Activated partial thromboplastin time (APTT) experiment: mix 90 μL lamb plasma with 10 μL sample dissolved in normal saline, add 100 μL preheated APTT reagent, incubate at 37°C for 5 minutes, add 100 μL preheated 0.25 mol / L Calcium chloride, and start timing, record APTT time.

[0031] Prothrombin time (PT) experiment: Mix 90 μL lamb plasma with 10 μL sample dissolved in normal saline, incubate at ...

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Abstract

The present invention provides low molecular weight imitated stichopus japonicus selenka glycosaminoglycans, a preparation method, and applications thereof. Structural formula of the low molecular weight imitated stichopus japonicus selenka glycosaminoglycan is as follows: the skeletal structure is repetition structures of -4GlcUA beta 1-3GalNAc beta 1-disaccharide, and the O-4 / 6 site of acetyl aminogalactose is sulfated. Every disaccharide unit has a fucose branched chain, the fucose is 2, 4-bis substituted by sulphate groups, the possible connection sites are O-3 site (R) of glucuronate, or O-4 site (R1) or O-6 site (R2) of acetyl aminogalactose. M=7-12. The low molecular weight imitated stichopus japonicus selenka glycosaminoglycans with weight average molecular weight (Mw) in the range of 7000 to 12,000 accounts for 95%. The purity is more than 98%. The low molecular weight imitated stichopus japonicus selenka glycosaminoglycans can be applied to anticoagulation and antithrombotic therapy, can reduce bleeding risk at the same time, and provides method support for the development of holothurian glycosaminoglycan products with similar structures.

Description

technical field [0001] The invention relates to the field of anticoagulant and antithrombotic drugs, in particular to a low-molecular-weight japonicus japonicus glycosaminoglycan and its preparation method and application. Background technique [0002] Sea cucumber has always been a valuable nourishing food, known as "sea ginseng". Traditional Chinese medicine believes that: sea cucumber tastes dry and salty, nourishes the kidney, benefits the essence, stimulates urination, strengthens impotence and treats impotence. Moreover, studies have shown that sea cucumbers can improve memory, delay gonad aging, prevent arteriosclerosis, diabetes and anti-tumor effects. Sea cucumbers contain various nutrients that are beneficial to the human body, such as proteins, amino acids, sea cucumber polysaccharides, vitamins, minerals, and sea cucumber saponins. Imitation sea cucumber ( Apostichopus japonicus ) is a kind of edible sea cucumber belonging to the phylum Echinodermata, the class...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/726A61P7/02
Inventor 吕志华王远红杨洁
Owner OCEAN UNIV OF CHINA