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Ilex asprella root polysaccharides and application thereof to preparation of anticomplement drugs

A technology of polysaccharides and polysaccharides, which is applied to polysaccharides, natural homogeneous polysaccharides in Chinese medicinal materials, polysaccharides, and the preparation of anti-complement drugs, which can solve the problems of research reports and anti-complement activities of polysaccharides in plums that have not yet been seen.

Active Publication Date: 2017-12-12
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Modern pharmacological researches have shown that genus genus has obvious antipyretic, analgesic, anti-inflammatory, and immune-regulating effects, but so far, there has been no research report on the anti-complement activity of genus genus and its polysaccharides

Method used

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  • Ilex asprella root polysaccharides and application thereof to preparation of anticomplement drugs
  • Ilex asprella root polysaccharides and application thereof to preparation of anticomplement drugs
  • Ilex asprella root polysaccharides and application thereof to preparation of anticomplement drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Preparation of Gangmega root polysaccharide GDS1, GDS2 and GDS3

[0025] 19Kg of Gangmei root medicinal material is pulverized, refluxed and extracted with 95% ethanol for 3 times, the ethanol in the filter residue is evaporated, the filter residue is extracted with hot water for 3 times, filtered, the combined extracts are concentrated to an appropriate volume, and 4 times the volume of Anhydrous ethanol, let it stand for about 3 hours, filter to obtain a precipitate, redissolve the precipitate in water, add trichloroacetic acid to make the concentration 15% to remove free protein, centrifuge, and adjust the supernatant to neutral with 1.0mol / L NaOH , concentrated to an appropriate volume, dialyzed with flowing water for 3 days with a dialysis membrane with a molecular weight cut-off of 3000, concentrated the dialysate to an appropriate volume, and obtained about 15.8 g of fluffy crude polysaccharide after freeze-drying. Weigh 12.0 g of crude polysaccharide, ...

Embodiment 2

[0029] Structural characterization of embodiment 2 ganglion polysaccharides (GDS1, GDS2 and GDS3)

[0030] (1) Determination of molecular weight

[0031] Using TSK-GEL GMPWXL gel column (300×7.6mm), the mobile phase is 0.01mol / LNaCl, the flow rate is 0.8mL / min, the column temperature is 25°C, GDS1, GDS2 and GDS3 are accurately calculated by Wyatt eighteen-angle laser light scattering The molecular weights are 92.77KDa, 16.22KDa and 21.84KDa, respectively.

[0032] (2) Elemental analysis results

[0033] GDS1: C, 40.30%; H, 6.68%; N, 0.56%.

[0034] GDS2: C, 37.80%; H, 5.88%; N, 0.57%.

[0035] GDS3: C, 29.91%; H, 5.45%; N, 0.67%.

[0036] (3) Specific rotation and CD spectrum

[0037] GDS1: CD: 202nm (-4.594); GDS2: CD: 201nm(-2.242), 223nm(1.882); GDS3: CD: 194nm(1.882), 230nm(0.993), 209nm(-2.158)

[0038] (4) Determination of total sugar, uronic acid and protein content

[0039] The total sugar content of GDS1 determined by sulfuric acid-phenol method was 95....

Embodiment 4

[0054] Example 4 Anti-complement alternative pathway test in vitro

[0055] Take complement (human serum) 0.2mL, add AP diluent (barbital buffer, pH=7.4, containing 5mMMg 2+ , 8mM EGTA) was prepared as a 1:1 solution, and double-diluted into 1:2, 1:4, 1:8, 1:16, 1:32, 1:64 and 1:128 solutions. Take 0.15mL of complement, 0.15mL of AP diluent and 0.20mL of 0.5% rabbit erythrocytes (RE), mix well, place in a low-temperature high-speed centrifuge at 37°C for 30min, and centrifuge at 5000rpm and 4°C for 10min. Take 0.20 mL of the supernatant from each tube and place it in a 96-well plate, and measure the absorbance at 405 nm. A full hemolysis group (0.20mL 0.5% RE dissolved in 0.3mL triple distilled water) was also set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Plot the dilution of complement on the X-axis and the percentage of hemolysis on the Y-axis. The ...

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Abstract

The invention belongs to the field of traditional Chinese medicine, and relates to three homogenous polysaccharides in a water extract of ilex asprella roots and application thereof to preparation of anticomplement drugs. The three ilex asprella root homogenous polysaccharides GDS1, GDS2 and GDS3 are isolated from ilex asprella roots, a commonly used heat-clearing and detoxifying traditional Chinese medicine in Lingnan. In vitro test results show that the prepared ilex asprella root polysaccharides GDS1, GDS2 and GDS3 have an obvious inhibitory effect on hemolysis caused by activation of classical and alternative pathways of a complement, and have a remarkable inhibiting effect on activated classical pathways and alternative pathways of the complement, wherein CH50 values are 0.134+ / -0.024 mg / mL, 0.187+ / -0.031 mg / mL and 0.098+ / -0.011 mg / mL respectively, and AP50 values are 0.297+ / -0.037 mg / mL, 0.312+ / -0.033 mg / mL and 0.195+ / -0.030 mg / ML respectively. The ilex asprella root polysaccharides can be used as an active ingredient to further prepare natural anti-complement drugs.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicines and relates to polysaccharides, in particular to the natural homogeneous polysaccharides in the Chinese medicinal material Gangmei and its application in preparing anti-complement drugs. Background technique [0002] Respiratory system virus infection (such as SARS, H1N1 and other viral infections) can easily cause acute lung injury, and then lead to acute respiratory distress syndrome (ARDS), which is not only highly contagious, but also has a high fatality rate and is extremely harmful. Studies have shown that the pathogenesis of the disease can trigger excessive activation of the complement system and produce a large number of biologically active complement products. These complement components can cause blood vessels to dilate, release inflammatory mediators, and promote systemic inflammatory responses, resulting in tissue damage. Therefore, the industry believes that inhibiting th...

Claims

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Application Information

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IPC IPC(8): A61K31/715A61K31/702A61P7/00C07H3/06C07H1/08C08B37/00
CPCA61K31/702A61K31/715A61K36/185C07H1/08C07H3/06C08B37/006A61K2300/00
Inventor 陈道峰卢燕温泉
Owner FUDAN UNIV
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