High-molecular weight Cordyceps militaris polysaccharide, its preparation method and its application in the preparation of anti-complement drugs

A technology of Cordyceps militaris polysaccharide and weight-average molecular weight is applied in directions such as pharmaceutical combinations, medical preparations containing active ingredients, pharmaceutical formulations, etc., to achieve the effect of significant complement inhibitory activity

Active Publication Date: 2021-10-22
YANBIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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, its preparation method and its application in the preparation of anti-complement drugs
  • High-molecular weight Cordyceps militaris polysaccharide, its preparation method and its application in the preparation of anti-complement drugs
  • High-molecular weight Cordyceps militaris polysaccharide, its preparation method and its application in the preparation of anti-complement drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Cordyceps militaris polysaccharide CMP-1, CMP-2 and CMP-3

[0027] Take 2.5 kg of Cordyceps militaris medicinal material and pulverize it, extract three times with 95% ethanol, filter to get the dregs, and dry to constant weight. Weigh the dried Cordyceps militaris residue, extract at a liquid-to-solid ratio of 35:1 (mL / g) at an extraction temperature of 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 a final concentration of 85%. After standing at 4°C for 24 hours, the supernatant was removed by centrifugation, and the precipitate was dried to constant weight to obtain the crude polysaccharide of Cordyceps militaris. Wet column packing is used, the pretreated DEAE-52 cellulose is loaded into the chromatography column, the crude polysaccharide of Cordyceps militaris is redissolved with deionized water, and the sample is loaded, followed by deionized wa...

Embodiment 2

[0028] Structural characterization of embodiment 2 Cordyceps militaris polysaccharide (CMP-1, CMP-2 and CMP-3)

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

[0030]The homogeneity and molecular weight of polysaccharides from Cordyceps militaris were determined by high performance gel permeation chromatography with refractive index detector (HPGPC-RID). Pass 2.0 mg / mL 20 μL of Cordyceps militaris polysaccharide solution through a 0.45 μm microporous membrane, and then inject it into a Shodexsugar KS-804 sugar column (8.0 mm×300 mm). The chromatographic conditions are: ultrapure water is the mobile phase, the flow rate is 1.0 mL / min, the column temperature is 50°C, and the temperature of the differential refraction detector is 35°C. Data processing was recorded on a N2000 GPC chromatography workstation. A series of dextran with different weight-average molecular weights were used as standard products to make a calibration curve to calculate the molecular weigh...

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 15 min as the source of complement for this bypass pathway. Take 2% rabbit erythrocytes with GVB-Mg 2+ / EGTA buffer to 5×10 8 After cells / mL, NHSP and different concentrations of diluted Cordyceps militaris polysaccharides were added, pre-incubated for 15 min, rabbit red blood cells (Erab) were added and incubated in a 37°C water bath for 30 min, cooled on ice, terminated, and centrifuged. The supernatant of each tube was diluted in a 96-well plate, and the absorbance value was measured at 412nm. At the same time, the blank group, the full hemolysis group, the NHSP group, the negative drug control group (glucose) and the positive drug control group (heparin) were set up in the experiment. . Calculate the hemolysis inhibition rate of Cordyceps militaris polysaccharide under different concentration gradients. And use GraphPad...

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Abstract

The invention belongs to the technical field of polysaccharide drugs, and in particular relates to three uniform high-molecular-weight polysaccharides in Cordyceps militaris, a preparation method thereof, and an application in preparing anti-complement drugs. The present invention separates and obtains three homogeneous high molecular weight polysaccharides CMP-1, CMP-2 and CMP-3 from Cordyceps militaris. Experiments confirm that these homogeneous high molecular weight polysaccharides have significant inhibitory effect on complement activation, and can be further used as active Components for the preparation of novel anti-complement drugs.

Description

technical field [0001] The invention belongs to the technical field of polysaccharide drugs, and in particular relates to three uniform high-molecular-weight polysaccharides in Cordyceps militaris, a preparation method thereof, and an application in preparing anti-complement drugs. 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 elimination in the elimination of foreign pathogenic microorganisms and metabolism of apoptotic cells, and plays a role in the activation and correct guidance 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; studies have shown that, In multiple organ failure syndromes, such a...

Claims

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Application Information

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Patent Type & Authority Patents(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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