Water-soluble low-molecular-weight cordyceps polysaccharide with anti-tumor activity, preparation method and application thereof

A water-soluble, low-molecular technology, applied in the direction of anti-tumor drugs, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve the lack of regular scientific basis, chemical composition, chain structure, molecular size and anti-tumor effect Less research, complex structure and other issues

Inactive Publication Date: 2010-08-18
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Cordyceps polysaccharide is an anti-tumor active substance with a high content in Cordyceps sinensis, but there are many types of polysaccharides and complex structures. The current research preliminarily believes that the biological activity of polysaccharides is related to its structure, but the chemical composition, chain structure, and molecular size of Cordyceps polysaccharides There are few studies on the relationship between anti-tumor effects, and there is still a lack of regular scientific basis

Method used

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  • Water-soluble low-molecular-weight cordyceps polysaccharide with anti-tumor activity, preparation method and application thereof
  • Water-soluble low-molecular-weight cordyceps polysaccharide with anti-tumor activity, preparation method and application thereof
  • Water-soluble low-molecular-weight cordyceps polysaccharide with anti-tumor activity, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Embodiment 1: Preparation of water-soluble low-molecular-weight Cordyceps polysaccharide

[0010] Using artificial fermentation and cultivation of Cordyceps mycelium as raw material, Cordyceps mycelium powder → 30% ethanol solution 70 ℃ hot reflux extraction → centrifugation, suction filtration → retain the clear liquid, repeated extraction of the residue → combined extracts → concentration → freeze-drying → Crude polysaccharide CPS → degreasing → protein removal by sevage method → ​​centrifugation → retain the supernatant → add ethanol to make the concentration reach 50% → centrifuge to obtain a precipitate, and the precipitate is subjected to DEAE-Sephadex A-25 column chromatography → phenol sulfuric acid method tracking detection → merge near the peak Eluent→freeze drying→obtaining solid→passing Sephadex G-75 column chromatography→obtaining refined polysaccharide.

Embodiment 2

[0011] Example 2: Anti-tumor activity test of water-soluble low-molecular-weight Cordyceps polysaccharide detected by MTT reduction method

[0012] 1. Cell culture

[0013] MFC cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum, 100 μg / mL penicillin and streptomycin, 5% CO 2 , 37°C, under the condition of saturated humidity for routine culture, replace the fresh medium every 48h to 72h, and subculture at a ratio of 1:3 to ensure that the cells used in the experiment are in the logarithmic growth phase.

[0014] MFC cells were routinely cultured, and fresh medium was replaced every 48 hours, and Hanks solution (without Ga 2+ , Mg 2+ Plasma) After washing the cells twice, add 1-2 mL of trypsin (containing 0.05% trypsin and 0.02% EDTA) to cover the bottom of the culture flask, digest in the incubator for 10-15 minutes, add 3-4 mL of medium containing serum to stop digestion , collect the cell liquid and centrifuge (1000rpm / min, 7.5min), pour off the supe...

Embodiment 3

[0019] Embodiment 3: In vivo anti-tumor activity experiment

[0020] Mice were injected intravenously on the second day after being inoculated with Sarcoma 180 tumor cells. For 10 consecutive days, the various manifestations, tumor growth and death of the mice were observed. Tumor inhibition rate, the results showed: ① 25, 50, 100 mg / kg had inhibitory effects on the growth of S180 tumors, respectively 41.2, 51.5, and 70.1%, suggesting that the water-soluble low-molecular-weight Cordyceps polysaccharide has a strong anti-tumor effect.

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Abstract

The invention discloses water-soluble low-molecular-weight polysaccharide with anti-tumor activity for artificially cultured cordyceps mycelium, a preparation method and an application thereof. 30 percent ethanol is used as extractant, and the best extraction process of the polysaccharide has the temperature of 70 DEG C, material-liquid ratio of 1 to 20, and extracting time of 2 hours. The polysaccharide is subjected to grease removal, protein removal, 50 percent ethanol precipitation and 70 percent ethanol precipitation to prepare crude polysaccharide after freezing and drying. The crude polysaccharide is purified grade by grade through a DEAE Sephadex A-25 column and a Sephadex G-75 column to prepare the water-soluble low-molecular-weight polysaccharide. The polysaccharide has higher purity, wherein the polysaccharide content is 94.57 percent, and relative molecular mass is about 9,874. The water-soluble polysaccharide has obvious inhibiting effect on proliferation of liver cancer cell line Bel-7204 and gastric cancer cell line SGC-7901 of human beings, and has obvious inhibiting effect on growth of S180 tumor cells implanted in a mouse body. The water-soluble low-molecular-weight cordyceps polysaccharide has clear active components and controllable quality, and can be used as an anti-tumor medicament or a health-care product for improving body immunity.

Description

technical field [0001] The invention relates to a cordyceps polysaccharide, and at the same time, the invention also relates to a preparation method of the cordyceps polysaccharide and its application in anti-tumor and improving body immunity. Background technique [0002] Cordyceps polysaccharide is an anti-tumor active substance with a high content in Cordyceps sinensis, but there are many types of polysaccharides and complex structures. The current research preliminarily believes that the biological activity of polysaccharides is related to its structure, but the chemical composition, chain structure, molecular size of Cordyceps polysaccharides There are few studies on the relationship between it and anti-tumor effect, and there is still a lack of regular scientific basis. Contents of the invention [0003] The object of the present invention is to provide a polysaccharide from Cordyceps sinensis, which has clear active ingredients, clear structure, clear molecular weig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A61K31/715A61P35/00A61P37/04A61K36/068
Inventor 朱振元司传领钟玥如原静刘安军
Owner TIANJIN UNIV OF SCI & TECH
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