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Novel fucosylated chondroitin sulfate FCS hm and preparation method and application thereof

A chondroitin sulfate and fucosification technology is applied in the field of biomedicine to achieve the effects of good anticoagulant and antithrombotic activity, easy industrial production and simple operation.

Active Publication Date: 2019-01-22
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for better quality products made from enzylosed cartilage glycosaminogens obtained during chewing or baking processes. It also includes various techniques like filtrations and ionexchange resins to remove unwanted substances such as sugars and proteins while maintaining their structure integrity. These technical benefits make this type of material more effective than previous methods used before making them.

Problems solved by technology

This patented describes how certain types of carboxylic sugars called fucoidan have various beneficial properties like binding specific proteins involved in immune response pathways, ability to reduce platelets' activation after they come into contact with activating agents, promoting clotting reactions, reducing inflammations caused by viruses, enzyme production, suppressing lipid accumulations associated with cardiac disease, controlling bleeding outcomes, protecting against cancer spreads, improving drug delivery systems targeting Toll-like Receptor 4(Toll Like Receptors-4), and other applications related thereto including use in animal models.

Method used

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  • Novel fucosylated chondroitin sulfate FCS hm and preparation method and application thereof
  • Novel fucosylated chondroitin sulfate FCS hm and preparation method and application thereof
  • Novel fucosylated chondroitin sulfate FCS hm and preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0040] Example 1: Novel fucosylated chondroitin sulfate FCS hm preparation of

[0041] The novel fucosylated chondroitin sulfate FCS provided by the present invention hm , the FCS hm from Holothuria Mexicana Sea cucumber, its molecular weight is 50~200 kDa, the mass ratio of sulfate radical is 20~40%, and described FCS hm The main chain is composed of chondroitin sulfate A (CSA) and chondroitin sulfate E (CSE), and its fine structure is analyzed by multi-dimensional nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry, and it is found that it is mainly composed of four structural fragments: two The species are α-Fuc-2S4S and α-L-Fuc-4S by O -3 is linked to glucuronic acid (GlcA), the other two are α-L-Fuc-4S and α-L-Fuc-3S4S through O -6 is attached to acetylgalactosamine (GalNAc), and FCS hm The side chain contains 1~5 Fuc, the molar ratio of the structural fragment with 1 Fuc side chain is 40~80%, and the molar ratio of the structural ...

Embodiment 2

[0048] Example 2: Novel fucosylated chondroitin sulfate FCS hm structural representation of

[0049] (1) Determination of absolute molecular weight and purity analysis: the FCS was determined by high performance gel permeation chromatography (HPGPC)-multi-angle laser scattering method (MALLS) hm absolute molecular mass and purity.

[0050] The FCS that will be produced hm 0.1 mol L for sample -1 Na 2 SO 4 Dissolved to make 5 mg·mL -1 aqueous solution. Chromatographic analysis conditions: Shodex OHpak SB-804 HQ and Shodex OHpak SB-802.5 HQ are used in series, and the mobile phase is 0.1 mol L -1 Na 2 SO 4 Aqueous solution, the injection volume is 100 μL, the column temperature is 35°C, and the flow rate is 0.6 mL min -1 , the acquisition time is 45min, and the detector is a differential detector combined with a multi-angle laser scattering instrument. Experimental results such as figure 2 As shown, the differential and laser signals both present a single symmetr...

Embodiment 3

[0055] Example 3: Novel fucosylated chondroitin sulfate FCS hm Anticoagulation

[0056] (1) Determination of anticoagulant activity: The activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) were measured using a coagulation analyzer and a kit purchased from Nanjing Jiancheng Biotechnology Company. Such as Figure 10 As shown in A and 10B, compared with the negative control, FCS hm The sample can significantly prolong the APTT and TT of plasma, but has no activity on PT, indicating that FCS exerts anticoagulant activity through the intrinsic coagulation pathway, and this effect is dose-dependent, and its activity is similar to that of low molecular weight heparin. In addition, CSA and CSE standard products have no anticoagulant activity, which proves that their fucose side chain is very important for their anticoagulant effect.

[0057] (2) Exploration of the anticoagulant mechanism: Experiments were carried out on 96-well plates using t...

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Abstract

The invention provides a novel fucosylated chondroitin sulfate FCS hm and a preparation method and an application thereof. The fucosylated chondroitin sulfate has the molecular weight of 50-200 kDa and has the sulfate content of 20-40%; a main chain of the fucosylated chondroitin sulfate is mainly composed of chondroitin sulfate A and chondroitin sulfate E. The FCS hm is mainly composed of four structural fragments, and a side chain contains 1-5 fucoses. The FCS hm exhibits good anticoagulant and antithrombotic activities through an endogenous coagulation pathway, is similar to low-molecular-weight heparin, and has inhibitory effects on FIIa and FXa mediated by antithrombin III. In addition, the FCS hm has high affinity with fibroblast growth factors FGF1 and FGF2, and has anti-angiogenesis effect. The preparation process of the compound provided by the invention has the advantages of low cost, simple operation and easy industrialized production.

Description

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Claims

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Application Information

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Owner OCEAN UNIV OF CHINA
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