Search

CN-121975036-A - Poria cocos uniform polysaccharide and preparation method and application thereof

CN121975036ACN 121975036 ACN121975036 ACN 121975036ACN-121975036-A

Abstract

The invention discloses a poria cocos uniform polysaccharide and a preparation method and application thereof. The invention combines anion exchange column chromatography technology and molecular sieve technology to separate and purify the pachyman from the poria cocos, and in vitro cell researches show that the pachyman has strong immune activation effect on macrophages, can enhance phagocytic effect of the macrophages, and provides a foundation for research and development of pachyman health function products.

Inventors

  • CHEN WEIDONG
  • ZHANG YUE
  • WANG YANYAN
  • Pan Xiuyi
  • Zhang Ningxiao
  • WANG MENGYA

Assignees

  • 安徽中医药大学

Dates

Publication Date
20260505
Application Date
20260318

Claims (9)

  1. 1. The preparation method of the poria cocos uniform polysaccharide is characterized by comprising the following steps of: (1) Drying and pulverizing Poria sclerotium into uniform powder, extracting with water and precipitating with ethanol to obtain Poria crude extract; (2) Removing protein and micromolecular impurities from the crude extract of poria cocos through a Sevag method and dialysis to obtain crude poria cocos polysaccharide; (3) Separating and purifying crude pachyman by adopting DEAE-52 cellulose column and SEPHACRYL S HR gel column to obtain pachyman.
  2. 2. The method for preparing pachyman according to claim 1, wherein in the step (1), the water extraction ratio is 1:8-12, the extraction temperature is 50-70 ℃, the extraction time is 1-2h, and the extraction times are 2-3 times.
  3. 3. The method for preparing pachyman according to claim 1, wherein in the step (3), DEAE-52 cellulose column is used for separation and purification, specifically, after crude pachyman solution is loaded, ultrapure water, naCl solution of 0.3M, 0.6M, 0.9M and 1M are sequentially used as eluent for stepwise elution, the fractions eluted by ultrapure water and 0.3M NaCl solution are collected, and PCP-W and PCP-S polysaccharide are obtained after concentration and freeze drying, respectively.
  4. 4. The method for preparing pachyman according to claim 1, wherein in the step (3), SEPHACRYL S HR gel column is used for separation and purification, specifically, PCP-W polysaccharide is loaded, ultrapure water is used as eluent for elution, concentration and dialysis are combined, and pachyman PCP-W1 is obtained after freeze drying.
  5. 5. A pachyman homogeneous polysaccharide prepared by the preparation method of any one of claims 1 to 4.
  6. 6. The pachymaran according to claim 5, wherein the pachymaran has a chain structure of → 2, 6) - α -D-Galp- (1 → 6) - β -D-Glcp- (1 → 3) - β -D-Glcp- (1 → and → 3, 6) - β -D-Glcp- (1 → residues constituting a main chain and branching sites located at the C-2 and → 3, 6) - α -D-Galp- (1 → C-6 positions of → 2, 6) - β -D-Glcp- (1 → and the pachymaran has a weight average molecular weight of 18.3 kDa, mainly composed of galactose, glucose, mannose and fucose.
  7. 7. Use of a pachyman according to claim 5 or 6 for the preparation of an immunomodulatory drug.
  8. 8. The use according to claim 7, wherein the medicament comprises the pachymaran according to claim 5 or 6 and a pharmaceutically acceptable carrier.
  9. 9. The medicament according to claim 8, wherein the dosage form of the medicament is a tablet, a capsule, a granule, a pill or an oral liquid.

Description

Poria cocos uniform polysaccharide and preparation method and application thereof Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to a poria cocos uniform polysaccharide, a preparation method and application thereof. Background Poria is dry sclerotium of Polyporaceae fungus Poria cocos (Schw.) Wolf, and is one of the well-known ten Anhui medicines, and has pharmacological effects of enhancing organism immunity, resisting tumor, protecting liver and resisting inflammation. And has the advantages of low toxic and side effects, high biological activity and the like, and has higher medicinal value. Polysaccharide is the main chemical composition of Poria cocos, and its total sugar content is about 80% of dry weight of Poria cocos. Research shows that the medicinal fungus polysaccharide has the function of activating macrophage, can regulate the secretion of cytokines such as interleukin, interferon, tumor necrosis factor superfamily, colony stimulating factor, chemotactic factor, growth factor and the like, and has the function of immunoregulation. The structural characteristics of polysaccharide such as space three-dimensional structure, monosaccharide composition, sugar residue composition and molecular weight are important substance structural basis for exerting pharmacological activity. Different structural features may cause different pharmacologically active responses. At present, the research on pachyman is mainly focused on extraction and separation, primary structure analysis, traditional pharmacological effects and the like. There is a delicate hunting in terms of the higher structure analyse and structure-activity relationship, which severely restricts the scientific research and product development of pachyman. Disclosure of Invention Aiming at the problems in the prior art, the invention aims to provide a poria cocos uniform polysaccharide and a preparation method and application thereof. The invention is realized by the following technical scheme: A preparation method of pachyman comprises the following steps: (1) Drying and pulverizing Poria sclerotium into uniform powder, extracting with water and precipitating with ethanol to obtain Poria crude extract; (2) Removing protein and micromolecular impurities from the crude extract of poria cocos through a Sevag method and dialysis to obtain crude poria cocos polysaccharide; (3) Separating and purifying crude pachyman by adopting DEAE-52 cellulose column and SEPHACRYL S HR gel column to obtain pachyman. In the step (1), the water extraction liquid ratio is 1:8-12, the extraction temperature is 50-70 ℃, the extraction time is 1-2h, and the extraction times are 2-3 times. In the step (3), DEAE-52 cellulose column is adopted for separation and purification, specifically, after crude pachyman solution is loaded, ultrapure water, 0.3M, 0.6M, 0.9M and 1M NaCl solution are sequentially used as eluent for step elution, the components eluted by the ultrapure water and the 0.3M NaCl solution are collected, and PCP-W and PCP-S polysaccharide are respectively obtained after concentration and freeze drying. In the step (3), SEPHACRYL S HR gel column is adopted for separation and purification, specifically, PCP-W polysaccharide is loaded, ultrapure water is used as eluent for elution, concentration and dialysis are combined, and the pachymaran PCP-W1 is obtained after freeze drying. The invention also provides a poria cocos uniform polysaccharide, which is prepared by the preparation method. The invention provides a chain structure of poria cocos uniform polysaccharide, wherein the chain structure of 2, 6) -alpha-D-Galp- (1- & gt, 6) -beta-D-GlcP- (1- & gt, 3) -beta-D-GlcP- (1- & gt and 3, 6) -beta-D-GlcP- (1- & gt residues form a main chain, branch sites are positioned at C-2 and C-6 positions of 2, 6) -alpha-D-Galp- (1- & gt, 3, 6) -beta-D-GlcP- (1- & gt, and the weight average molecular weight of the poria cocos uniform polysaccharide is 18.3 kDa and mainly comprises galactose, glucose, mannose and fucose. The invention also provides an application of the poria cocos uniform polysaccharide in preparing an immunoregulatory medicament. The medicine provided by the invention comprises the poria cocos uniform polysaccharide and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers described herein include, but are not limited to, lubricants, fillers, binders, disintegrants, pH adjusters, surfactants, antioxidants, and the like, in amounts conventional in the art. The pharmaceutical dosage form of the invention can be prepared according to the conventional method in the field, and the pharmaceutical dosage form is tablets, capsules, granules, pills or oral liquid and the like. Compared with the prior art, the invention has the following advantages: The invention combines anion exchange column chromatography technology and molecular sieve technology to separate and purify the pachyma