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A kind of low molecular weight imitation sea cucumber glycosaminoglycan and its preparation method and application

A sea cucumber glycosaminoglycan and low-molecular-weight technology, which can be used in blood diseases, extracellular fluid diseases, drug combinations, 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, etc., to prolong blood coagulation The effect of enzyme time, high product purity, and simple and convenient method

Active Publication Date: 2017-12-05
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 to 45000Da; and the patent CN101057859A utilizes 5% H 2 o 2 Low molecular weight sea cucumber glycosaminoglycans with a molecular weight of 8000-12000 were prepared by degradation of 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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  • A kind of low molecular weight imitation sea cucumber glycosaminoglycan and its preparation method and application
  • A kind of low molecular weight imitation sea cucumber glycosaminoglycan and its preparation method and application
  • A kind of low molecular weight imitation sea cucumber glycosaminoglycan and its preparation method and application

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Embodiment 1

[0024] Example 1: Preparation of low molecular weight imitation sea cucumber glycosaminoglycan

[0025] Dissolve 10mg of sea cucumber glycosaminoglycan (HG, weight average molecular weight about 98kDa) in 1mL of 100mmol / L K containing 50mmol / L hydrogen peroxide 2 HPO 4 In the solution, the pH value of the solution is 7.5. 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, 4000rpm Centrifuge for 20 minutes to remove the pellet. 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 mol...

Embodiment 2

[0029] Implementation example 2: In vitro anticoagulant activity experiment of low molecular weight imitation sea cucumber glycosaminoglycan

[0030] 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.

[0031] A. In vitro anticoagulation experiment of low molecular weight imitation sea cucumber glycosaminoglycan

[0032]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.

[0033] 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 invention provides a low-molecular-weight imitation sea cucumber glycosaminoglycan and its preparation method and application. Its structural formula is as follows:. Its skeleton structure is →4GlcUAβ1→3GalNAcβ1→disaccharide repeating structure, and the O-4 / 6 position of acetylgalactosamine is sulfated. There is a fucose branch on each disaccharide unit, which is 2,4-disulfate-substituted fucose, and the possible connection site is the O-3 position (R) of glucuronic acid, or the acetylamino group O‑4 position (R1) or O‑6 position (R2) of galactose. m=7~12. The weight average molecular weight (Mw) of the low molecular weight imitation japonicus glycosaminoglycans in the range of 7000-12000 accounted for 95%. Its purity is greater than 98%. The low-molecular-weight imitation sea cucumber glycosaminoglycan prepared by the invention can be applied to anticoagulant and antithrombotic treatment, and can reduce the risk of bleeding at the same time, and provides sufficient method support for the development of sea cucumber 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. Apostichopus japonicus is an edible sea cucumber belonging to the phylum Echinodermata, the class Holothuria, the order Apostichopus...

Claims

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Application Information

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