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Cedar polysaccharide, its preparation method and its application in the preparation of anti-complement drugs

A cedar polysaccharide and complement technology, applied in the preparation of anti-complement drugs, five natural homogeneous polysaccharides in cedar and its preparation field

Active Publication Date: 2021-09-07
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Based on the basis of the prior art and no reports on the separation and preparation of homogeneous polysaccharides with anti-complement activity in Cedar cypress, the inventors of the present application tried to strengthen the research on the anti-complement active components of related traditional Chinese medicines, so as to provide substances for the treatment of complement-related diseases The basis, specifically providing cedar polysaccharide and its preparation method and its use in the preparation of anti-complement drugs

Method used

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  • Cedar polysaccharide, its preparation method and its application in the preparation of anti-complement drugs
  • Cedar polysaccharide, its preparation method and its application in the preparation of anti-complement drugs
  • Cedar polysaccharide, its preparation method and its application in the preparation of anti-complement drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of cedar polysaccharides XB-PS2, XB-PS3, XB-PS4, XB-PS5 and XB-PS6

[0026] Cedar herb 3Kg is crushed, extracted with 95% ethanol, filtered, the dregs are extracted 3 times with aqueous solution, concentrated, centrifuged, and 4 times the volume of 95% ethanol is added to the supernatant, left to stand, centrifuged to remove the supernatant, the precipitate is washed with water Redissolve, recover under reduced pressure, remove ethanol; reconstitute the solution with trichloroacetic acid to remove free protein, centrifuge, adjust the supernatant to neutral, dialyze, concentrate, freeze-dry to obtain crude polysaccharide, add 100g of crude polysaccharide to dissolve in distilled water, centrifuge , the supernatant was initially separated by DEAE-cellulose column chromatography, eluted with distilled water, 0.1, 0.4, 0.8, 1.6 and 2.0mol / L NaCl solution, the elution volume was greater than 2 times the column volume (about 3L), and the flow rate was 10...

Embodiment 2

[0033] The structural characterization of embodiment 2 cedar polysaccharides (XB-PS2, XB-PS3, XB-PS4, XB-PS5 and XB-PS6)

[0034] (1) Determination of molecular weight

[0035] 18-angle laser light scattering gel chromatography system is used to detect molecular weight. The basic principle is that homogeneous polysaccharides pass through gel permeation chromatography to form symmetrical chromatographic peaks, and form light scattering after being irradiated by 18-angle laser light. The light scattering signal is directly related to the molecular weight. . The data is calculated with Astar (version 5.3.1) software and directly gives the molecular weight;

[0036] Experimental method: Accurately weigh 5.0 mg of homogeneous polysaccharide, make it into a 10 mg / ml solution, and pass through a 0.45-micron microporous membrane before sample injection. Chromatographic conditions: flow rate 0.5mg / ml, injection volume 20μl, 0.1% NaCl solution as mobile phase, column temperature 25°C,...

Embodiment 4

[0058] Example 4 Alternative Pathway Complement Inhibition Test

[0059] Serum was taken from healthy adult male volunteers and mixed with VBS-Mg-EGTA buffer (barbital buffer, pH=7.4, containing 5mM Mg 2+and 8mM EGTA) diluted 1:8 as a source of complement for the alternative pathway. Rabbit red blood cells stored in 3.8% sodium citrate solution were prepared into 0.5% rabbit red blood cells with VBS-Mg-EGTA buffer solution. Accurately weigh about 3 mg of polysaccharide, add VBS-Mg-EGTA buffer, and dilute to 8 concentrations in double. 150 μL of polysaccharide solutions of different concentrations and 150 μL of 1:8 complement were pre-incubated at 37°C for 10 minutes, then 200 μL of 0.5% rabbit red blood cells were added, placed in a low-temperature high-speed centrifuge after 30 minutes in a water bath at 37°C, and centrifuged at 5000 rpm and 4°C for 10 minutes . Take 200 μL supernatant from each tube in a 96-well plate, and measure the absorbance at 405 nm. At the same ti...

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Abstract

The invention belongs to the technical field of traditional Chinese medicine, and relates to five homogeneous polysaccharides in cedarwood, a preparation method thereof and an application in preparation of anti-complement drugs. The present invention separates and obtains five homogeneous polysaccharides XB‑PS2, XB‑PS3, XB‑PS4, XB‑PS5 and XB‑PS6 from Cedar Cedar, a traditional Chinese medicine for clearing away heat and detoxification, and experiments prove that these Cedars homogeneous polysaccharides have significant effect on complement activation. The inhibitory effect can be further used as an active ingredient to prepare new anti-complement drugs.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicine, and relates to polysaccharides, a preparation method thereof, and applications in pharmacy, in particular to five natural homogeneous polysaccharides from cedarwood, a preparation method thereof, and an application in preparing anti-complement drugs. Background technique [0002] The prior art discloses that the complement system is an important part of the human immune system, and its normal activation plays an important role in eliminating foreign microorganisms, removing damaged or dead cells and tissues in the body, and maintaining the balance of the body. However, excessive activation of this system will cause excessive response of the human immune system, resulting in damage to the normal tissues of the human body, such as rheumatoid arthritis, senile dementia, systemic lupus erythematosus (SLE) and rejection after organ transplantation. In multiple organ failure syndromes, such ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61K31/715A61P37/02A61P19/02A61P29/00A61P11/00A61P31/12A61P13/12A61P1/16
CPCA61K31/715A61P1/16A61P11/00A61P13/12A61P19/02A61P29/00A61P31/12A61P37/02C08B37/0003C08B37/006
Inventor 卢燕陈道峰夏龙朱孟夏
Owner FUDAN UNIV
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