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Radix arnebiae polysaccharides and application thereof to preparation of anti-complement drugs

A technology of comfrey polysaccharide and application, applied in the field of natural homogeneous polysaccharide, preparation of anti-complement drugs, and polysaccharide, can solve the problems such as separation, preparation and activity report of homogeneous polysaccharide with anti-complement activity 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

But so far, there is no report on the isolation, preparation and activity of a homogeneous polysaccharide that has anti-complement activity in Comfrey

Method used

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  • Radix arnebiae polysaccharides and application thereof to preparation of anti-complement drugs
  • Radix arnebiae polysaccharides and application thereof to preparation of anti-complement drugs
  • Radix arnebiae polysaccharides and application thereof to preparation of anti-complement drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of comfrey polysaccharides APS-1 and APS-2

[0022] 20Kg of comfrey is crushed, extracted by cold immersion in 95% ethanol, filtered, extracted 3 times with hot water, filtered, combined extracts, concentrated, centrifuged, added 4 times the volume of 95% ethanol to the supernatant, let stand, Centrifuge to remove the supernatant, redissolve the precipitate in water, recover under reduced pressure, and remove ethanol; then use trichloroacetic acid to remove free protein from the reconstituted solution, centrifuge, adjust the supernatant to neutral, dialyze, concentrate, and freeze-dry to obtain crude polysaccharide. Crude polysaccharide 0.5 g was dissolved in distilled water, centrifuged, and the supernatant was divided into DEAE-cellulose columns (Cl -1 type, 30×2.5 cm) chromatography for preliminary separation. Elute with distilled water, 0.4, 0.8, 1.2 and 2.0 mol / L NaCl solutions, the elution volume is greater than 2 times the column volume (a...

Embodiment 2

[0024] Example 2 Structural characterization of comfrey polysaccharides (APS-1 and APS-2)

[0025] (1) Molecular weight determination

[0026] APS-1 and APS-2 were compared with standard T-series dextran, and the results showed that the molecular weights of APS-1 and APS-2 were both less than 2×106 Da, but greater than 1×106 Da.

[0027] (2) Elemental analysis results

[0028] The elemental analysis results of APS-1 are: C, 42.35%; H, 6.59%; N, 1.84%.

[0029] APS-2 element analysis results are: C, 30.96%; H, 4.46%; N, 1.50%.

[0030] (3) Specific rotation

[0031] APS-1 specific curl: [ α ] D 25 =+14.2 ( c 0.318, H 2 O).

[0032] APS-2 specific curl: [ α ] D 25 = +24.7 ( c 0.205, H 2 O).

[0033] (4) Determination of total sugar, uronic acid, protein and sulfate content

[0034] The total sugar content of APS-1 determined by sulfuric acid-phenol method was 95.36%; the total sugar content of APS-2 was 95.81%.

[0035] The content of uronic acid in APS-1 was ...

Embodiment 3

[0046] Example 3 Anti-complement classical pathway test in vitro

[0047] Take 0.1ml of complement (guinea pig serum), add BBS to prepare a 1:5 solution, and double-dilute with BBS to 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1: 640 solution. Dissolve 0.1 ml of 1:1000 hemolysin, each concentration of complement and 2% SRBC in 0.3 ml BBS, mix well, put in a low-temperature high-speed centrifuge at 37 ºC for 30 min, and centrifuge at 5000 rpm and 4 ºC for 10 min. Take 0.2 ml of the supernatant from each tube in a 96-well plate, and measure the absorbance at 405 nm. A full hemolysis group (0.1 ml 2% SRBC dissolved in 0.5 ml 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. Taking the dilution of complement as the X-axis, the percentage of hemolysis caused by each dilution of complement is plotted as the Y-axis. The lowest com...

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Abstract

The invention belongs to the field of traditional Chinese medicine, and relates to active ingredients with the anti-complement function in natural medicine, in particular to natural homogeneous polysaccharides in radix arnebiae and application thereof to preparation of anti-complement drugs. The two homogeneous polysaccharides APS-1 and APS-2 are separated from radix arnebiae. Experiments confirm that the homogeneous polysaccharides in radix arnebiae have a significant inhibiting effect on the classical pathway and the bypass pathway for complement activation, wherein the CH50 and AP50 values of the radix arnebiae polysaccharide APS-1 for inhibiting the classical pathway and the bypass pathway of a complement system are 203+ / -20 mug / ml and 45+ / -8 mug / ml respectively, and the CH50 and AP50 values of the radix arnebiae polysaccharide APS-2 for inhibiting the classical pathway and the bypass pathway of the complement system are 282+ / -11 mug / ml and 144+ / -17 mug / ml respectively. The natural homogeneous polysaccharides in radix arnebiae can further serve 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 polysaccharides, in particular to the natural homogeneous polysaccharides in comfrey and its use in the preparation of anti-complement drugs. Background technique [0002] The prior art discloses that the normal activation of the complement system 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, the abnormal activation of this system will cause the excessive reaction of the human immune system, cause damage to the normal tissues of the human body, and participate in the pathological process of various diseases. Studies have shown that chronic diseases related to excessive activation of complement include rheumatoid arthritis, senile dementia, systemic lupus erythematosus (SLE) and rejection after organ transplantation; Overactivation of complement also ...

Claims

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Application Information

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IPC IPC(8): A61K31/715A61P29/00A61P19/02A61P25/28A61P37/02A61P37/06A61P9/10A61P11/00
CPCA61K31/715A61K36/30
Inventor 陈道峰卢燕金家宏
Owner FUDAN UNIV
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