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Lily leaf polysaccharide and its preparation method and use in the preparation of anti-complement medicine

A technology of polysaccharide and lily, applied in the direction of drug combination, pharmaceutical formula, plant raw material, etc., can solve the problems that have not been seen before.

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

Crude polysaccharides from the leaves of Sapura chinensis have obvious inhibitory effects on the classical and alternative pathways of complement activation, CH 50 and AP 50 78±5μg / mL and 110±9μg / mL respectively, but so far there is no report on the isolation and preparation of homogeneous polysaccharides with anti-complement activity in the leaves of S.

Method used

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  • Lily leaf polysaccharide and its preparation method and use in the preparation of anti-complement medicine
  • Lily leaf polysaccharide and its preparation method and use in the preparation of anti-complement medicine
  • Lily leaf polysaccharide and its preparation method and use in the preparation of anti-complement medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of polysaccharides from the leaves of Sapura chinensis BL-PS1, BL-PS3, BL-PS4, BL-PS5 and BL-PS6

[0026] 3Kg of the herb leaves of Ficus praecox were crushed, extracted with 95% ethanol, filtered, the dregs were extracted 3 times with aqueous solution, concentrated, centrifuged, and 4 times the volume of 95% ethanol was added to the supernatant, left to stand, centrifuged to remove the supernatant, precipitated with Redissolve in water, recover under reduced pressure, and remove ethanol; the reconstituted solution is then removed with trichloroacetic acid to remove free protein, centrifuged, and the supernatant is adjusted to neutrality, dialyzed, concentrated, and freeze-dried to obtain crude polysaccharide. 100 g of crude polysaccharide was dissolved in distilled water, centrifuged, and the supernatant was preliminarily separated by a DEAE-cellulose chromatographic column. Elute with distilled water, 0.1, 0.4, 0.8, 1.6 and 2.0 mol / L NaCl solutio...

Embodiment 2

[0033] Example 2 Structural Characterization of Polysaccharides from Fructus Lilii Leaf (BL-PS1, BL-PS3, BL-PS4, BL-PS5 and BL-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 temper...

Embodiment 4

[0059] Example 4 Alternative Pathway Complement Inhibition Test

[0060] 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 to 1:8, as the complement source of the bypass pathway; rabbit red blood cells stored in 3.8% sodium citrate solution were configured into 0.5% rabbit red blood cells with VBS-Mg-EGTA buffer solution; accurately weighed polysaccharide about 3 mg, add VBS-Mg-EGTA buffer solution, double-dilute to 8 concentrations, 150 μL of polysaccharide solution of different concentrations and 150 μL of 1:8 complement are pre-incubated at 37°C for 10 minutes, add 200 μL of 0.5% rabbit red blood cells, and incubate at 37°C After bathing in water for 30 minutes, put it into a low-temperature high-speed centrifuge, and centrifuge 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...

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Abstract

The invention belongs to the field of traditional Chinese medicines, and relates to five homogeneous polysaccharides in leaves of Sapura chinensis, a preparation method thereof and an application in preparation of anti-complement drugs. The present invention separates and obtains five homogeneous polysaccharides BL-PS1, BL-PS3, BL-PS4, BL-PS5 and BL-PS6 from the heat-clearing and detoxifying traditional Chinese medicine Saplings sativa leaves. Experiments confirm that these homogeneous polysaccharides activate the classical pathway and peripheral pathway of complement. Both pathways have significant inhibitory effects, and can be further used as active ingredients to prepare novel anti-complement 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 pharmacy; in particular, it relates to five natural homogeneous polysaccharides in the leaves of Sapura chinensis, 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 an overreaction 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; Studies ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61P37/02A61K31/715
CPCA61K31/715A61K36/60A61P37/02C08B37/0003C08B37/006
Inventor 陈道峰夏龙卢燕
Owner FUDAN UNIV
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