Total cedar polysaccharide and application of total cedar polysaccharide in preparation of immunosuppressive drug

A technology of total polysaccharide and cedar, applied in the field of preparing anti-complement and anti-inflammatory drugs

Active Publication Date: 2017-08-25
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported in the literature that cedar contains volatile oils, flavonoids, phenylpropanoids, st...

Method used

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  • Total cedar polysaccharide and application of total cedar polysaccharide in preparation of immunosuppressive drug
  • Total cedar polysaccharide and application of total cedar polysaccharide in preparation of immunosuppressive drug
  • Total cedar polysaccharide and application of total cedar polysaccharide in preparation of immunosuppressive drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 Preparation of Cedar Cedar Total Polysaccharides

[0015] Grind the cedar branches and leaves, and use 95% ethanol to heat and reflux under reduced pressure at a ratio of 1:30 to liquid (55°C, 3 times, 4 hours each time). Ratio of material to liquid is subjected to decompression heating and reflux extraction (70°C, divided into 3 times, each time for 5h). Concentrate the water extract to 0.4g crude drug / ml, add 4 times the volume of 95% ethanol, filter after alcohol precipitation for 24h, and dilute with water. The total polysaccharides were obtained by freeze-drying after precipitation and reconstitution; according to this process, three batches of total polysaccharides of cedar cedar SP1, SP2 and SP3 were prepared respectively.

Embodiment 2

[0016] Example 2 Structural characterization of cedarwood total polysaccharides

[0017] (1) Molecular weight distribution:

[0018] Cedar cypress total polysaccharide SP3 was formulated into 0.3mg / ml aqueous solution for HPLC analysis: TSK-GEL GMPWXL gel column (300×7.6 mm) was used, the mobile phase was deionized water, the flow rate was 0.8 mL / min, and the column temperature was 25°C , the detection wavelength is 210nm; the polysaccharide standard is analyzed by the same method, and the comparison of the spectrum shows that the molecular weight of cedar polysaccharide is basically less than 25000 Da;

[0019] (2) Monosaccharide composition:

[0020] Take 8 mg polysaccharide sample and add 2ml of 1M TFA, heat at 110°C for 5h, centrifuge, evaporate the supernatant to dryness, and wash the precipitate three times with methanol; 0.3 M NaOH, heated at 70°C for 2h for hydrolysis. After cooling, add 1ml 0.3M HCl to neutralize, add chloroform to extract 3 times, discard the chlo...

Embodiment 3

[0033] Example 3 Determination of anti-complement activity of total polysaccharides of cedar cypress by cell hemolysis

[0034] Take about 2.0mg total polysaccharides of cedar cedar, accurately weighed, dissolve in 0.8ml BBS (barbiturate buffer solution), and double-dilute to obtain 2.5, 1.25, 0.625, 0.3125, 0.1563, 0.0781, 0.0391, 0.0195mg / Each 0.4ml of sample solution in 1ml was drawn into 0.2ml to 0.4ml of BBS as a blank control; 0.2ml of complement, 0.1ml of hemolysin and 0.1ml of 2% sheep red blood cell solution were added to the remaining sample solution, and centrifuged Then take the supernatant and measure the absorbance at 405nm, and calculate the IC of the hemolysis inhibition rate of the three batches of cedarwood polysaccharide after deducting the absorbance of the blank control 50 SP1: 25.04μg / ml, SP2: 35.40μg / ml, SP3: 23.82μg / ml respectively; IC of positive control drug heparin sodium 50 It was 27.81 μg / ml.

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Abstract

The invention belongs to the field of traditional Chinese medicine and relates to a total cedar polysaccharide. The polysaccharide is a water-soluble crude polysaccharide extracted and prepared from Sabina pingii var.wilsonii branches and leaves, and the structural characteristics such as sugar content, uronic acid content, protein content, molecular weight distribution and monosaccharide composition of the polysaccharide are represented. Activity determination results indicate that the cedar polysaccharide has an obvious anticomplementary action, the half effective inhibition concentration of a complement activation classical pathway is up to 23-35 micro-g/ml, and the polysaccharide is equivalent to a positive control drug in heparin sodium activity. In addition, the level of mouse peritoneal macrophage cell secretion NO under LPS stimulation can be remarkably inhibited, and it is indicated that the total cedar polysaccharide has a very good anti-inflammatory effect. The total cedar polysaccharide can be used for preparing anti-complement, anti-inflammatory and other immunosuppressive related drugs.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicine, and relates to a plant polysaccharide and its medicinal use, in particular to a total polysaccharide extracted and prepared from cedar (Sabina pingii var. wilsonii) and its use in preparing anti-complement and anti-inflammatory 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 overreaction of the human immune system, cause damage to the normal tissues of the human body, and participate in the pathological process of various diseases such as rheumatoid arthritis, systemic lupus erythematosus, and acute respiratory distress syndrome. Currently, immunosuppressants such as glucocorticoids, cyclophosphamide, a...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A61P29/00A61P37/06
Inventor 卢燕陈道峰朱孟夏凌丽君张庆琳
Owner FUDAN UNIV
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