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Sabina tibetica polysaccharides as well as preparation method and application thereof to preparation of anticomplement drugs

A technology of juniper polysaccharides and complements, applied in the preparation of anti-complement drugs, four natural homogeneous polysaccharides in juniper juniper and their preparation, can solve the problem that has not yet been seen

Active Publication Date: 2019-11-26
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The study found that the crude polysaccharide of Juniper juniper had significant anti-complement activity, CH 50 and AP 50 76±9μg / ml and 87±4μg / ml, respectively, but there is no report on the isolation and preparation of homogeneous polysaccharides with anti-complement activity in Juniper japonica

Method used

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  • Sabina tibetica polysaccharides as well as preparation method and application thereof to preparation of anticomplement drugs
  • Sabina tibetica polysaccharides as well as preparation method and application thereof to preparation of anticomplement drugs
  • Sabina tibetica polysaccharides as well as preparation method and application thereof to preparation of anticomplement drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Preparation of Juniper polysaccharide YB-PS2, YB-PS3, YB-PS4 and YB-PS5

[0025] Grind 3Kg of Juniper juniper, extract with 95% ethanol, filter, extract the dregs with aqueous solution for 3 times, concentrate, centrifuge, add 4 times the volume of 95% ethanol to the supernatant, let it stand, centrifuge to remove the supernatant, and precipitate Reconstitute with water, recover under reduced pressure, remove ethanol; reconstitute the solution with trichloroacetic acid to remove free protein, centrifuge, adjust the supernatant to neutral, dialyze, concentrate, and freeze-dry to obtain crude polysaccharide. 100g of crude polysaccharide was dissolved in distilled water, centrifuged, and the supernatant was initially separated by DEAE-cellulose chromatography, and eluted with distilled water, 0.1, 0.4, 0.8, 1.6 and 2.0mol / L NaCl solution, and the elution volume was greater than 2 times the column volume (about 3L), the flow rate is 10mL / min, collect each fraction...

Embodiment 2

[0030] Example 2 Structural characterization of juniper polysaccharides (YB-PS2, YB-PS3, YB-PS4 and YB-PS5)

[0031] (1) Determination of molecular weight

[0032] 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;

[0033] Experimental method: Accurately weigh 5.0mg of homogeneous polysaccharide, configure it into a 10mg / ml solution, pass it through a 0.45 micron microporous membrane before injection, chromatographic conditions: flow rate 0.5mg / ml, injection volume 20μl, 0.1% NaCl solution As the mobile phase, the column temperature is 25°C, the la...

Embodiment 4

[0053] Example 4 Alternative Pathway Complement Inhibition Test

[0054] 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 erythrocytes stored in 3.8% sodium citrate solution were prepared into 0.5% rabbit erythrocytes with VBS-Mg-EGTA buffer solution, accurately weighed about 3 mg of polysaccharide, added VBS-Mg-EGTA buffer solution, and double-diluted to 8 concentrations 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 10min. Take 200 μL of supernatant from each tube in a 96-well plate, and measure the absorbance at 405 nm. At the same time, ...

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Abstract

The invention belongs to the technical field of traditional Chinese medicines and relates to four homogeneous polysaccharides in Sabina tibetica as well as a preparation method and application of thefour homogeneous polysaccharides to preparation of anticomplement drugs. The four homogeneous polysaccharides YB-PS2, YB-PS3, YB-PS4 and YB-PS5 are separated from Sabina tibetica serving as a traditional Chinese medicine capable of clearing away heat and toxic materials, and proved by experiments, the homogeneous polysaccharides have an obvious effect on inhibiting complement activation and can befurther used as active components to prepare novel anticomplement drugs.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicine, and relates to a polysaccharide, a preparation method thereof and an application in preparation thereof, in particular to four natural homogeneous polysaccharides in juniper juniper, a preparation method thereof and an application in preparation of anti-complement drugs. Background technique [0002] 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 as ische...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A61P37/02
CPCA61K31/715A61P37/02C08B37/0003C08B37/006
Inventor 卢燕陈道峰夏龙朱孟夏
Owner FUDAN UNIV
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