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BCG-polysaccharide and preparation and analysis identification methods thereof

A BCG polysaccharide and polysaccharide technology, applied in analytical materials, material separation, instruments, etc., can solve the problems of complexity, reduced drug action selectivity, increased adverse reactions, etc., achieving simple and efficient operation, low equipment requirements, and enhanced body The effect of resistance

Active Publication Date: 2016-03-02
JIUZHITANG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the complexity of the components of BCG-PSN reduces the selectivity of drug action and increases the number of adverse reactions; the active ingredients are not clear, which brings certain difficulties to the research of drug quality control, the development of new preparations, the structure-activity relationship and the interpretation of the mechanism of action. Restricts the optimization and innovation of BCG drugs, affecting its medicinal value and market competitiveness among similar drugs
There are no reports on the structure of BCG polysaccharides at home and abroad, which brings challenges to the interpretation of the structure-activity relationship and mechanism of action of BCG polysaccharides

Method used

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  • BCG-polysaccharide and preparation and analysis identification methods thereof
  • BCG-polysaccharide and preparation and analysis identification methods thereof
  • BCG-polysaccharide and preparation and analysis identification methods thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation of embodiment 1 BCG polysaccharide

[0034] Take 300 mg of BCG polysaccharide nucleic acid sample and dissolve it in deionized water, pre-treat and equilibrate with 0.5 mol / L NaCl solution 3 times the column volume to obtain a DEAE-32 cellulose column (2.5*40cm), and absorb the sample for 5 hours. Elute 3 column volumes with deionized water, collect the eluate, concentrate to an appropriate amount, dialyze with a dialysis bag (molecular weight cut-off 3000Da) for 72 hours, and change the deionized water every 5 hours. The solution in the dialysis bag was concentrated and freeze-dried at 70°C to obtain a white solid polysaccharide with a yield of 34.2%. The obtained BCG polysaccharide was used for the following experimental analysis.

[0035] Physical and chemical properties and structural analysis

[0036] Properties: white solid, easily soluble in water.

[0037] Ultraviolet absorption: no absorption at 200-800nm, and strong absorption peak at 630nm ...

Embodiment 2

[0047] Example 2 Effect of BCG polysaccharide on T lymphocyte subsets and cytokines in immunocompromised model mice

[0048] 1. Establishment of immunocompromised model mice

[0049] The mouse model of immunocompromised function was completed with reference to "Guiding Principles of Pharmacodynamics of Anti-inflammatory and Immunological Drugs. Compilation of Guiding Principles for Preclinical Research of New Drugs (Western Medicine) (Pharmacology, Pharmacology and Toxicology)", that is, daily injection of hydrocortisone (100 mg / kg , ic, 0.1ml / 10g body weight), once a day for a total of 7 days.

[0050] 2. Experimental grouping

[0051] Test is divided into 4 groups: normal animal control group, immunocompromised model group, BCG polysaccharide injection group (concentration=1.0mg / ml, polysaccharide content 96%), commercially available BCG polysaccharide nucleic acid injection group (concentration= 1.0mg / ml, polysaccharide content ≈75%, nucleic acid content ≈15%).

[0052] Th...

Embodiment 3

[0066] Example 3 Effect of BCG polysaccharide on guinea pig genital herpes simplex virus (HSV-2) infection

[0067] 1. Establishment of animal models

[0068] For the modeling method, see (BernateinDI, etal.JInfectDis, 2001; 183(6):844), that is, first wash the vulva with normal saline, rub the vagina several times with a dry cotton swab, causing vaginal mucosal damage, and then use a syringe connected to a gastric lavage needle Absorb HSV-2 (potency is 10 -4～-6 TCID 50 , provided by the Wuhan Institute of Virology, Chinese Academy of Sciences) 0.1ml, insert the needle into the vagina of the guinea pig about 3-4cm, inject the virus into the vaginal fornix and withdraw slowly, stuff it into the vagina with a gelatin sponge to maintain the venom, a little venom on the needle drips on the vulva, Gently spread evenly with a glass rod to allow the virus to penetrate the skin.

[0069] 2. Grouping of animals

[0070] After injecting virus 14 days, according to skin lesion strati...

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Abstract

The invention discloses a BCG-polysaccharide and a preparation method and an analysis identification method thereof. The preparation method includes using BCG-PSN as raw material, using DEAE-32 cellulose chromatographic column to perform elution by using deionized water as an eluant according to physical property and chemical property differences of polysaccharide and nucleic acid, collecting an eluate, performing dialysis and desalting, performing condensation, and performing freeze drying to obtain BCG-polysaccharide with molecular weight of 1.78*10<4>Da. The analysis identification method includes performing complete acid hydrolysis and methylation of the BCG-polysaccharide, identifying the BCG-polysaccharide by means of GC-MS, IR, and NMR. The BCG-polysaccharide is a glucan having a main chain of (1->4)-alpha-D-Glcp and containing a few branches of ->4,6)-alpha-D-Glap-(1->, the BCG-polysaccharide has one branch in average eight main chain residue, and the main chain has a deoxidized methyl doublet on the position C6. The preparation method is simple, high-efficient, and without damage, and the BCG-polysaccharide obtained by separating and purifying has high product purity. The structural characterization of the BCG-polysaccharide has important meaning on the researches of effective component and structure-activity relationship of BCG-polysaccharide.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and in particular relates to a BCG polysaccharide and its preparation and structure characterization method. technical background [0002] As one of the material bases of life activities, polysaccharides contain rich biological information. They come from a wide range of sources, have complex structures with large differences, and have diverse biological activities. They have extremely important roles and application prospects in life science research. Studies have shown that polysaccharides and their derivatives, such as many plant and fungal polysaccharides, have significant pharmacological activities in terms of immune regulation, anti-tumor, anticoagulant, and blood lipid reduction. They have been favored by domestic researchers in recent years, but the relevant bacterial polysaccharides research is still relatively small. [0003] BCG-PSN (Polysaccharidenucleic acid fraction of Bacillus Ca...

Claims

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Application Information

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IPC IPC(8): C08B37/00G01N30/02
Inventor 刘韶谷陟欣许雄博辛秀叶惠煊
Owner JIUZHITANG
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