Integrated peptidoglycan and preparation method thereof

A peptidoglycan and a complete technology, applied in the field of pharmaceutical preparations, can solve problems such as lack of Lactobacillus bulgaricus extraction, and achieve the effects of suitable large-scale production, anti-tumor effect and good effect

Active Publication Date: 2013-03-06
内蒙古双奇药业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there is a lack of reports on the extraction of intact peptidoglycan from Lactobacillus bulgaricus in relevant literature at home and abroad

Method used

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  • Integrated peptidoglycan and preparation method thereof
  • Integrated peptidoglycan and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The teichoic acid of Lactobacillus bulgaricus is extracted by trichloroacetic acid to increase the permeability of the cell membrane, which is conducive to the better action of external substances on the cells and the discharge of cell contents, and then degreases to remove the fatty substances on the cell membrane, and uses enzymatic hydrolysis The complete peptidoglycan obtained by removing impurities in the cell after centrifugation and freeze-drying can be seen through transmission electron microscope observation that the cell wall skeleton structure is complete. Lactobacillus bulgaricus can be obtained from the public sale of Inner Mongolia Shuangqi Pharmaceutical Co., Ltd.

[0030] The preparation method of the complete peptidoglycan comprises the following steps:

[0031] (1) Cleaning and collection steps of bacteria:

[0032] Take 100 grams of Lactobacillus bulgaricus wet cells, add 1000 milliliters of normal saline, separate by centrifugation, centrifuge at 80...

Embodiment 2

[0045] Example 2: Structural integrity detection of a complete peptidoglycan of Lactobacillus bulgaricus disclosed by the present invention

[0046] The original thalline and the prepared complete peptidoglycan in Example 1 of the present invention are detected by electron microscopy, see figure 1 shown. The results showed that the complete peptidoglycan still retained the complete external structure of the original thalline.

Embodiment 3

[0047] Example 3: Anti-tumor effect test of a complete peptidoglycan of Lactobacillus bulgaricus disclosed by the present invention

[0048]Aseptically extract the peritoneal effusion from H22 liver cancer Kunming mice, trypan blue staining to check the survival rate of cancer cells, the peritoneal effusion tumor cells with a survival rate greater than 95% are adjusted to 1×107 cells / mL with serum-free medium . 40 normal Kunming mice were taken, half male and half male, and after adaptive feeding for 3 days, 2×106 cancer cells (0.2mL) were inoculated and injected subcutaneously in the right axilla of each mouse. One week later, the mouse liver cancer model was successfully established, and a tumor with a diameter of about 1 cm grew at the inoculation site. After modeling, the mice were randomly divided into 4 groups, 10 in each group, namely the control group (sterilized PBS1mL / d), the low-dose Lactobacillus bulgaricus intact peptidoglycan group (0.1mL / d), and the middle-dose...

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Abstract

The invention discloses a integrated peptidoglycan which can suppress tumor growth effectively and can be widely used in antitumor drugs due to the integrated spatial structure of the peptidoglycan extracted. The invention further discloses a method for extracting the integrated peptidoglycan, which comprises the following steps of: extracting teichoic acid to enhance membrane permeability, degreasing to remove fatty substances on the cell membranes, removing protein substances in the cells through an enzymatic solution, and finally centrifuging and cleaning to remove impurities and freeze-drying to obtain the whole peptidoglycan. The invention has the advantages that the cell wall has a whole structure since the integrated peptidoglycan is extracted, thereby having excellent antitumor effect. The preparation method has the advantages of strong practicability, simple operation, short period, simple and reasonable steps, low cost and high yield, and is suitable for large-scale production.

Description

Technical field: [0001] The invention relates to the technical field of pharmaceutical preparations, in particular to a chemical method for extracting complete peptidoglycan from Lactobacillus bulgaricus and a method thereof. Background technique: [0002] Cancer is one of the important diseases that threaten human life. The World Health Organization and the health departments of governments all regard conquering cancer as an important task. The research and development of new and efficient anti-tumor drugs not only has good economic benefits, but also brings huge social benefits. In general, anticancer drugs are very expensive due to the scarcity of raw materials or high technological content. Therefore, research and production of an anticancer drug with significant curative effect and low price is needed by society. [0003] Scholars at home and abroad have clearly clarified that Lactobacillus bulgaricus is a physiologically beneficial bacterium, which plays an important...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K2/00
Inventor 黄少磊龚虹刘彦民付艳茹张丽萍冯谦程超
Owner 内蒙古双奇药业股份有限公司
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