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Composition comprising bacillus calmette guerin polysaccharides and bacillus calmette guerin nucleic acids and use of preparing medicament thereof

A technology of BCG polysaccharide and BCG nucleic acid, which is applied in the direction of medical preparations containing active ingredients, drug combinations, anti-tumor drugs, etc., can solve the problems of slow effective effect and short effective duration, and achieve good safety and Effectiveness, anti-allergic immunomodulatory effects

Active Publication Date: 2013-03-06
HUNAN SIQI BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the data of the current research, the clinical application of this preparation shows the shortcomings of slow marked effect and short effective duration. Whether this ratio is appropriate and optimal has not been optimized through scientific methods

Method used

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  • Composition comprising bacillus calmette guerin polysaccharides and bacillus calmette guerin nucleic acids and use of preparing medicament thereof
  • Composition comprising bacillus calmette guerin polysaccharides and bacillus calmette guerin nucleic acids and use of preparing medicament thereof
  • Composition comprising bacillus calmette guerin polysaccharides and bacillus calmette guerin nucleic acids and use of preparing medicament thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The cultivation and harvest of embodiment 1 BCG

[0037] (1) Bacterial culture: Dissolve the strains preserved in liquid cryogenic temperature (BCG strain D2PB302 for the preparation of BCG in China, provided by the vaccine room of China National Institute for the Control of Pharmaceutical and Biological Products) at room temperature, and inoculate them in Potato Sutong medium, continuously at 37°C. Cultivate for 14-20 days; or culture at 37°C for 15 days, then transfer to improved liquid Sutong medium, and culture at 37°C for 14-20 days.

[0038] Wherein, the preparation method of potato Sutong culture medium can be:

[0039] Take and wash fresh potatoes (1), pierce them into cylinders with a piercer, and cut them into 4 cm long slopes with a knife;

[0040] Rinse the potato slopes with running drinking water for 1 hour;

[0041] Rinse the potato slant pieces with purified water;

[0042] Rinse the slant block with Sutong medium;

[0043] Take 20ml of Sutong cultur...

Embodiment 2

[0058] The preparation of embodiment two BCG polysaccharide, nucleic acid mixture

[0059] (1) Preparation method one: dialysis

[0060] 1. Cell crushing and thermal phenol treatment: add the cell collected in Example 1 into purified water at a ratio of 10:1, crush the cell with a tissue mash homogenizer (12000 rpm / min), 3 min × 3 times, and Crush the bacteria, then add hot phenol (60-65°C) equal to the broken bacteria suspension, and keep warm for 30 minutes to 1 hour while stirring at a low speed.

[0061] 2. Extraction of the BCG polysaccharide nucleic acid mixture: naturally precipitate the hot phenolic mixed solution for 1 to 10 days, absorb the supernatant, centrifuge in a tube centrifuge, and put the supernatant into a dialysis bag (cut-off molecular weight > 5000 Dal Dayton) in 100 times the volume of water for injection dialyzed for 1 to 10 days to remove phenol. An appropriate amount of ethanol is added to the dialyzed liquid to make the alcohol content 60-85%, and...

Embodiment 3

[0065] Embodiment three prepares BCG polysaccharide and BCG nucleic acid from BCG polysaccharide nucleic acid mixture

[0066] 1. Preparation of ion exchange chromatography column: 1L to 100L ion exchange packing (Q Sepharose TM XL) According to the method provided in the instruction manual, pack into the corresponding chromatographic column, rinse the chromatographic column with 2 to 10 times the column volume of water for injection, and then equilibrate the chromatographic column with 2 to 10 times the column volume of normal saline for injection ;

[0067]2. Separation of polysaccharides and nucleic acids from the BCG polysaccharide and nucleic acid mixture: 10 times the column volume of the BCG polysaccharide and nucleic acid mixture prepared in Example 2 was loaded, and at the same time began to collect the flow-through. After loading the sample, wash the column with 2 times the column volume of normal saline, collect the column wash solution, combine the flow-through s...

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Abstract

A composition comprising BCG polysaccharide and BCG nucleic acid is provided, wherein the polysaccharide accounts for 10% to 69% of the mass of the composition, and the nucleic acid accounts for 30% to 89% of the mass of the composition, the The composition is prepared by mixing said polysaccharide and said nucleic acid, wherein said polysaccharide and said nucleic acid are obtained from a BCG culture by extracting said polysaccharide and said a mixture of nucleic acids, and separating said polysaccharide and said nucleic acids from this mixture by ion exchange chromatography. Also provided is the use of the composition comprising the polysaccharide and the nucleic acid in the preparation of biological agents for treating or preventing microbial infections, tumors, and allergies.

Description

technical field [0001] The invention relates to a composition comprising BCG polysaccharide and BCG nucleic acid and the application of the composition. Background technique [0002] BCG is bovine attenuated tuberculosis, non-pathogenic, and immunogenic. It is one of the most widely used vaccines in the world to replace the initial infection of tuberculosis with BCG inoculation to obtain immunity against tuberculosis. one. [0003] In 1882, Robert successfully isolated Mycobacterium tuberculosis and determined the pathogenic bacteria of tuberculosis. In 1908, Calnette and Guerin of the Pasteur Institute in France successfully bred a strain of attenuated Mycobacterium bovis and named it Bacillus Calmette-Guerin (BCG). In 1921, Weill-Halle successfully applied the bacteria to the human body to prevent and treat tuberculosis. In 1971, the Changsha Research Group for the Prevention and Treatment of Tracheitis first used the dead BCG scratch method to prevent and treat chronic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/04A61K31/715A61K31/7052A61P31/00A61P35/00A61P37/00A61P37/08
CPCA61K31/7052A61K31/715A61K39/39A61P31/00A61P35/00A61P37/00A61P37/08
Inventor 宁云山关继峰
Owner HUNAN SIQI BIOPHARM
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