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Ganoderma lucidum beta-glucan and preparation method thereof and application thereof to preparing immunomodulatory drugs

A technology of glucan and glucan molecular weight, applied in the field of medicine, to achieve the effects of abundant raw material sources, high product purity, and promoting cell phagocytosis of neutral red

Pending Publication Date: 2018-12-11
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

None of the above 3 patents involves the research on the residue after water extraction

Method used

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  • Ganoderma lucidum beta-glucan and preparation method thereof and application thereof to preparing immunomodulatory drugs
  • Ganoderma lucidum beta-glucan and preparation method thereof and application thereof to preparing immunomodulatory drugs
  • Ganoderma lucidum beta-glucan and preparation method thereof and application thereof to preparing immunomodulatory drugs

Examples

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Embodiment 1

[0028] Example 1: Preparation of Ganoderma lucidum β-glucan

[0029] The preparation method of ganoderma lucidum β-glucan of the present invention specifically comprises the following steps:

[0030] (1) Hot water extraction to remove water-soluble polysaccharides: crush red Ganoderma lucidum through a 60-mesh sieve, add distilled water according to the mass ratio of material to liquid 1:10, heat and extract at 80°C twice, each time for 2 hours, remove water-soluble polysaccharides, and remove water-soluble polysaccharides from red Ganoderma lucidum. Drying of medicinal material residues;

[0031] (2) Extraction of crude β-glucan: Add the residue of the medicinal material obtained in step (1) to a 2M aqueous NaOH solution at a mass ratio of 1:10, heat and extract at 60°C for 1 hour, extract twice, and centrifuge (4500r / min, 20 min) combined to collect the supernatant, add a molar concentration of 1M HCl aqueous solution, neutralize the supernatant, and concentrate under vacu...

Embodiment 2

[0034] Example 2: Structural characterization of Ganoderma lucidum β-glucan

[0035] The structural characterization of Ganoderma lucidum β-glucan of the present invention specifically includes the following steps:

[0036] (1) Absolute molecular weight determination and purity analysis: The absolute molecular mass of Ganoderma lucidum β-glucan was determined and the purity was analyzed by high performance gel permeation chromatography (HPGPC)-octadecanine laser light scattering method (MALLs).

[0037] Chromatographic conditions: Column: Shodex OHPak SB804HQ column connected in series with Shodex OHPak SB802.5HQ column; mobile phase: 0.1mol L -1 Na 2 SO 4 Aqueous solution; Differential detector and 18-angle laser light scattering instrument are connected in series to detect on-line, measure the molecular weight of β-glucan; Sample is single symmetrical peak in GPC spectrum (such as figure 1 Shown, the peak after 30min is NaCl), the sample purity is very high, and its absol...

Embodiment 3

[0042] Example 3: Determination of Immunomodulatory Activity of Ganoderma lucidum β-glucan

[0043] The immunomodulatory activity assay of Ganoderma lucidum β-glucan of the present invention specifically includes the following process:

[0044] RAW264.7 cells were cultured, planted in 96-well plates, the number of cells per well was 20,000, and cultured with drugs for 24 hours. After 24 hours, discard the supernatant, wash the cells 3 times with PBS, add 100 μL of 0.075 wt% concentration to each well, incubate for 20 minutes, discard the supernatant, wash the cells 3 times with PBS, add 200 μL of cell lysate to each well, and measure each well at 540 nm. Well absorbance value. At the same time, the effect of the test substance on the proliferation of RAW264.7 cells was measured by MTT method, and the phagocytosis was expressed by the ratio of the absorbance of the RAW264.7 cell phagocytosis neutral red test to the absorbance of the cell proliferation test measured by the MTT ...

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Abstract

The invention belongs to the technical field of medicines and particularly relates to a ganoderma lucidum beta-glucan, a preparation method thereof and an application thereof to preparing immunomodulatory drugs. The molecular weight of the Ganoderma lucidum beta-glucan is 5-500 kDa; the monosaccharide composition comprises glucose and glucuronic acid; the ratio of the two monosaccharides is 30:1-4:1; the connection mode mainly comprises ->3) Glc (beta1-> and->6) Glc (beta1->; and the ratio of the two connection modes is 1:1-3:1. The preparation method comprises the steps of carrying out hot water extraction on ganoderma lucidum medicinal materials to remove water-soluble polysaccharides; extracting residue by using an alkali solution, neutralizing extract by using hydrochloric acid and concentrating; precipitating by using ethanol to obtain a crude polysaccharide; and purifying the crude polysaccharide by using anion exchange resin to finally obtain the high-purity beta-glucan. Provedby cell experiments, the beta-glucan disclosed by the invention is capable of significantly promoting the process of phagocytosing neutral red by RAW264.7 cells and can be used for preparing the immunomodulatory drugs.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a ganoderma lucidum beta-glucan, a preparation method thereof and an application in preparation of immunoregulatory medicines. Background technique [0002] Ganoderma lucidum polysaccharide is one of the more important active ingredients in Ganoderma lucidum. Structural analysis shows that polysaccharides of Ganoderma lucidum are mainly high molecular weight sugar polymers, mainly formed by monosaccharides such as glucose, xylose, mannose, galactose and fucose. Composition of sugar macromolecules. Ganoderma lucidum polysaccharides obtained from different sources and extraction methods have different characteristics such as monosaccharide composition, molecular weight, branched conformation, and solubility, and can induce various immune responses to exert immunomodulatory effects. [0003] The traditional extraction methods of Ganoderma lucidum polysaccharides mainly...

Claims

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Application Information

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IPC IPC(8): C08B37/02A61P37/02
CPCA61P37/02C08B37/0003C08B37/0087
Inventor 于广利顾菲菲蔡超郝杰杰李佳
Owner OCEAN UNIV OF CHINA
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