High-molecular-weight cordyceps militaris polysaccharide, preparation method thereof and application of high-molecular-weight cordyceps militaris polysaccharide in preparation of anticomplement drugs

A technology of Cordyceps militaris polysaccharide and weight average molecular weight, which is applied in the direction of drug combination, medical preparations containing active ingredients, and pharmaceutical formulas, etc.

Active Publication Date: 2020-12-15
YANBIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no report on the isolation and preparation of uniform high molecular weight polysaccharides with anti-complement activity in Cordyceps militaris

Method used

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  • High-molecular-weight cordyceps militaris polysaccharide, preparation method thereof and application of high-molecular-weight cordyceps militaris polysaccharide in preparation of anticomplement drugs
  • High-molecular-weight cordyceps militaris polysaccharide, preparation method thereof and application of high-molecular-weight cordyceps militaris polysaccharide in preparation of anticomplement drugs
  • High-molecular-weight cordyceps militaris polysaccharide, preparation method thereof and application of high-molecular-weight cordyceps militaris polysaccharide in preparation of anticomplement drugs

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 prepares Cordyceps militaris polysaccharide CMP-1, CMP-2 and CMP-3

[0027] Take 2.5kg of Cordyceps militaris material and crush, extract three times with 95% ethanol, filter to get the medicinal residue, and dry to constant weight. Weigh the dried Cordyceps militaris slag, take the liquid-solid ratio of 35:1 (mL / g), extract the temperature at 90° C. for 2 hours, and filter to obtain the filtrate. The filtrate was concentrated to 1 / 5 of the original volume, and 95% ethanol was added to the final concentration of 85%. After standing at 4°C for 24 hours, the supernatant was removed by centrifugation, and the precipitate was taken and dried to constant weight to obtain Cordyceps militaris crude polysaccharide. Using wet packing, the pretreated DEAE-52 cellulose was loaded into the chromatography column, the crude polysaccharide of Cordyceps militaris was redissolved in deionized water, and the samples were loaded, followed by deionized water, 0.1mol / L and 0.2m...

Embodiment 2

[0028] Embodiment 2 The structural characterization of Cordyceps militaris polysaccharides (CMP-1, CMP-2 and CMP-3)

[0029] (1) Determination of weight average molecular weight

[0030]The homogeneity and molecular weight of Cordyceps militaris polysaccharides were determined by high performance gel permeation chromatography combined with refractive index detector (HPGPC-RID). The 2.0 mg / mL 20 μL Cordyceps militaris polysaccharide solution was passed through a 0.45 μm microporous membrane, and then injected into a Shodexsugar KS-804 sugar column (8.0 mm×300 mm). The chromatographic conditions were as follows: ultrapure water was the mobile phase, the flow rate was 1.0 mL / min, the column temperature was 50°C, and the temperature of the differential refraction detector was 35°C. Data processing was recorded on an N2000 GPC chromatography workstation. The molecular weights of polysaccharides were calculated using a series of dextran with different weight average molecular weig...

Embodiment 4

[0050] Example 4 Alternative pathway complement inhibition test

[0051] Normal healthy human serum by GVB-Mg 2+ / EGTA buffer was incubated on ice for 15min as the source of complement for this alternative pathway. Take 2% rabbit erythrocytes with GVB-Mg 2+ / EGTA buffer to 5×10 8 After cells / mL, NHSP and different concentrations of Cordyceps militaris polysaccharide after dilution were added, pre-incubated for 15 min, and rabbit erythrocytes (Erab) were added and incubated in a 37°C water bath for 30 min, cooled on ice, terminated the reaction, and centrifuged. Dilute the supernatant from each tube in a 96-well plate, and measure the absorbance at 412 nm. At the same time, a blank group, a complete hemolysis group, an NHSP group, a negative drug control group (glucose) and a positive drug control group (heparin) were set up in the experiment. . The inhibition rate of hemolysis under different concentration gradients of Cordyceps militaris polysaccharide was calculated. An...

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Abstract

The invention belongs to the technical field of polysaccharide drugs, and particularly relates to three uniform high-molecular-weight polysaccharides in cordyceps militaris, a preparation method thereof and application of the high-molecular-weight polysaccharides in preparation of anticomplement drugs. The three uniform high-molecular-weight polysaccharides CMP-1, CMP-2 and CMP-3 are separated from the cordyceps militaris, and experiments prove that the uniform high-molecular-weight polysaccharides have a remarkable inhibition effect on complement activation and can be further used as active ingredients for preparing novel anticomplement drugs.

Description

technical field [0001] The invention belongs to the technical field of polysaccharide medicines, and particularly relates to three uniform high molecular weight polysaccharides in Cordyceps militaris, a preparation method thereof, and use in the preparation of anti-complement medicines. Background technique [0002] The prior art discloses that the complement system is the first line of defense of the body's natural immune defense, plays an important role in monitoring, defense and clearance in the elimination of foreign pathogenic microorganisms and apoptotic cell metabolism, and plays an important role in the activation and correct orientation of adaptive immunity. deep influence. The complement system is a "double-edged sword". Excessive activation can cause damage to the body's own normal tissues, such as rheumatoid arthritis, senile dementia, systemic lupus erythematosus, and rejection after organ transplantation. In multiple organ failure syndromes, such as ischemia-r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/715A61K36/068A61P7/00C08B37/00
CPCA61K31/715A61K36/068A61K2236/333A61K2236/39A61K2236/51A61K2236/53A61K2236/55A61P7/00C08B37/0003
Inventor 孙金凤胡政宇李镐周微王佳鸣金龙
Owner YANBIAN UNIV
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