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Fucosan sulphate, preparation method thereof, and application of fucosan sulphate in preparing anti-influenza virus medicine

A fucoidan sulfate, anti-influenza virus technology, applied in the directions of antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc., to achieve the effects of strong inhibitory effect, simple preparation process and high safety

Active Publication Date: 2014-06-25
威海人生药业有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestic and foreign patents have reported some patents on the preparation and application of fucoidan sulfate (publication number CN103288978A), but Kjellmaniella crassifolia, Laminaria saccharina or mozuku derived from brown algae (Nemacystus decipiens) fucoidan sulfate preparation and its research and application in anti-influenza A H1N1, H5N1 and H3N2 influenza viruses have not yet been reported

Method used

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  • Fucosan sulphate, preparation method thereof, and application of fucosan sulphate in preparing anti-influenza virus medicine
  • Fucosan sulphate, preparation method thereof, and application of fucosan sulphate in preparing anti-influenza virus medicine
  • Fucosan sulphate, preparation method thereof, and application of fucosan sulphate in preparing anti-influenza virus medicine

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preparation example Construction

[0037] The preparation method of fucoidan sulfate provided by the present invention comprises the following steps:

[0038] (1) Hot water extraction and dilute acid precipitation: extract brown algae powder after degreasing treatment with hot water, combine the extracts, precipitate with dilute hydrochloric acid, and centrifuge. The hot water extraction temperature is 70-90° C., and the pH of the dilute acid precipitation solution is 1-3.

[0039] (2) Calcium chloride precipitation: add 1 to 3 mol / L calcium chloride aqueous solution to the supernatant after (1) centrifugation, take the supernatant after centrifugation, concentrate, dialyze, precipitate with ethanol, and obtain fucoidan sulfate after drying .

[0040] (3) DEAE-Cellulose purification: Dissolve the fucoidan sulfate obtained in (2) in water, use sodium chloride aqueous solution as the mobile phase (0<sodium chloride concentration≤2mol / L), and undergo DEAE-Cellulose anion exchange Resin separation, sulfuric acid-...

Embodiment 1

[0043] Embodiment 1: Preparation of fucoidan sulfate ester

[0044] 1. Extract the defatted kelp powder in 80°C water bath for 3 to 5 times, combine the extracts, cool to room temperature, adjust the pH of the extracts to 1 to 3 with dilute hydrochloric acid, centrifuge, and collect the supernatant.

[0045] 2. Stir and add 3mol / L of CaCl to the supernatant prepared in step 1 2 Aqueous solution, until the precipitation no longer occurs, centrifuge to take the supernatant, transfer the supernatant to a dialysis bag with a molecular weight cut-off of 7000D, dialyze until the conductivity of the dialysis fluid no longer changes, concentrate the fluid in the dialysis bag under reduced pressure, and vacuum freeze-dry Get fucoidan sulfate (KCP).

[0046] 3. Dissolve the fucoidan sulfate prepared in step 2 with distilled water, then pass through DEAE-Cellulose anion exchange resin for further purification, and use NaCl aqueous solution (0<sodium chloride concentration≤2mol / L) as elu...

Embodiment 2

[0050] Embodiment 2: the effect of fucoidan sulfate against influenza A (H1N1) virus

[0051] 1. Fucoidan sulfate inhibits NA activity of H1N1, H5N1 and H3N2 influenza viruses

[0052] The fucoidan sulfate (KCP), H1N1, H5N1 and H3N2 neuraminidase NA and NA-specific substrate MUNANA (4-methylumbelliferyl-N-acetyl-α-neuralminic acid) were co-incubated in MES buffer, Using 4MU (7-hydroxy-4-methylcoumarin), the product of the interaction between NA and MUNANA, can produce 460nm fluorescence under the excitation of 355nm incident wavelength, and calculate the inhibition of fucoidan sulfate (KCP) on NA activity by detecting the change of fluorescence intensity Rate (the test method is recorded in the literature Li Hanbing, etc. Acta Pharmacina Sinica, 2009, 44(2): 162-166.), with zanamivir as the positive control drug.

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Abstract

The invention belongs to the field of marine medicines, and relates to a fucosan sulphate, a preparation method thereof and application of the fucosan sulphate in preparing an anti-influenza virus medicine. Polysaccharide with a main chain taking alpha-1,2-D-mannose and beta-1,4-D-glucuronic acid as repetitive units, and with a branched chain of alpha-1,3-L-fucosan sulphate is obtained by virtue of hot-water extraction, calcium chloride precipitation and DEAE-Cellulose chromatographic column purification. The fucosan sulphate prepared by the invention is high in inhibition effect on the influenza virus neuraminidase activities of A H1N1, H5N1 and H3N2, and obvious in protection effect on the dog kidney epithelial cells infected by A H1N1. The fucosan sulphate provided by the invention has the advantages of being rich in raw material source, simple in preparation process, easy to industrialize, high in product water solubility, high in stability, free from toxic and side effects, and the like, and has the prospect of being developed to the anti-influenza virus medicine.

Description

technical field [0001] The invention belongs to the field of marine medicines, and relates to a fucoidan sulfate extracted from brown algae Kjellmaniella crassifolia, Laminaria saccharina or Nemacystus decipiens, a preparation method thereof and preparation methods thereof. Application of anti-influenza virus drugs. Background technique [0002] Influenza A Virus (IAV) belongs to Orthomyxoviridae family (Orthomyxoviridae) and has an enveloped negative-strand RNA virus. It can cause acute respiratory infectious diseases. It has the characteristics of strong infectivity and high pathogenicity, and seriously endangers human health. . Due to the different structure and antigenicity of hemagglutinin protein (HA) and neuraminidase protein (NA) on the surface of influenza A virus, it can be divided into different subtypes, such as H1N1, H3N2 and H5N1. Since the outbreak of the H1N1 influenza virus that originated in Mexico in April 2009, different subtypes of influenza viruses su...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/737A61P31/16
Inventor 于广利吴建东王伟赵小亮吕友晶刘潇潇
Owner 威海人生药业有限公司
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