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Preparation method of mannuronate oligosaccharides, and application of mannuronate oligosaccharides in preparation of combined antibacterial agent for inhibiting drug-resistant bacteria

A technology of uronic acid oligosaccharides and antibacterial agents, which is applied in the field of medicine, can solve the problems of high cost, complicated operation, and low purity, and achieve the effects of increasing potency, simple preparation method, and reducing MIC value

Inactive Publication Date: 2019-02-01
HEBEI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] One of the purposes of the present invention is to provide a method for preparing mannuronic acid oligosaccharides, so as to solve the problems of complex operation, high cost and low purity in the existing mannuronic acid oligosaccharide preparation methods

Method used

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  • Preparation method of mannuronate oligosaccharides, and application of mannuronate oligosaccharides in preparation of combined antibacterial agent for inhibiting drug-resistant bacteria
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  • Preparation method of mannuronate oligosaccharides, and application of mannuronate oligosaccharides in preparation of combined antibacterial agent for inhibiting drug-resistant bacteria

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Experimental program
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Effect test

Embodiment 1

[0030] a. Take 15g of kelp, add 200mL of formaldehyde hydrochloric acid solution (10mL of formaldehyde plus 0.84mL of hydrochloric acid water to make up to 100mL, shake well), soak for 4h, wash with water, and filter with gauze. Add 600mL of 1% sodium carbonate solution to the filter residue, bathe in 60°C water for 3h, let cool, add 100mL of water, stir evenly, centrifuge at 4000rpm×5min, and take the supernatant. The pH of the supernatant was adjusted to 2.0 with hydrochloric acid and allowed to stand for 12 hours. Collect the gel by centrifugation at 4000rpm×5min. Add 200mL of 2% sodium hydroxide solution to the gel, stir to dissolve the gel completely, add 400mL of ethanol, mix well, and precipitate with alcohol to obtain a yellow-white gel-like precipitate (algin), which should be lyophilized and stored away from light.

[0031] b. Take 2g of alginate, add 400mL of water, and soak overnight. Add an appropriate amount of hydrochloric acid to make the concentration 0.2mol...

Embodiment 2

[0037] a, with embodiment 1.

[0038] b. Take 2g of alginate, add 400mL of water, and soak overnight. Add an appropriate amount of hydrochloric acid to make the concentration 0.2mol / L, heat in a water bath at 100°C and stir magnetically for 4 hours. After cooling, centrifuge to take the supernatant, adjust the pH of the supernatant to neutral after rotary evaporation, and freeze-dry to obtain crude LMOS.

[0039] c. Dissolve an appropriate amount of the above-mentioned LMOS in 800 μL of water, vortex and mix well, and centrifuge at 12000 rpm for 3 min. Take the centrifuged supernatant and put it on a Sephadex-25 dextran gel column (2.6 cm×30 cm), use water as the eluent, collect the eluate, about 3mL per tube, use the phenol-sulfuric acid method to detect sugar, and use nitric acid The silver test solution detects salt, and water is used as a control. Collect the salt-free and sugar-containing eluate, and freeze-dry to obtain the finished LMOS. The resulting finished produ...

Embodiment 3

[0041] a, with embodiment 1.

[0042] b. Take 2g of alginate, add 400mL of water, and soak overnight. Add an appropriate amount of hydrochloric acid to make the concentration 0.1mol / L, heat in a water bath at 100°C and stir magnetically for 2h. After cooling, centrifuge to take the supernatant, adjust the pH of the supernatant to neutral after rotary evaporation, and freeze-dry to obtain crude LMOS.

[0043] c. Dissolve an appropriate amount of the above-mentioned LMOS in 800 μL of water, vortex and mix well, and centrifuge at 12000 rpm for 3 min. Take the centrifuged supernatant and put it on a Sephadex-25 dextran gel column (2.6 cm×30 cm), use water as the eluent, collect the eluate, about 3mL per tube, use the phenol-sulfuric acid method to detect sugar, and use nitric acid The silver test solution detects salt, and water is used as a control. Collect the salt-free and sugar-containing eluate, and freeze-dry to obtain the finished LMOS. The resulting finished product is...

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Abstract

The invention provides a preparation method of mannuronate oligosaccharides, and an application of the mannuronate oligosaccharides in the preparation of a combined antibacterial agent for inhibitingdrug-resistant bacteria. The prepration method comprises the following steps: carrying out acid degradation on sodium alginate in a reaction system having an acid concentration of 0.1-1.0 mol / L in 100DEG C water bath for 1-4 h, and purifying the obtained product to obtain the mannuronate oligosaccharides having a polymerization degree focusing on 2-6. The application of the mannuronate oligosaccharides in the preparation of the combined antibacterial agent for inhibiting drug-resistant bacteria is characterized in that the mannuronate oligosaccharides and a traditional antibacterial agent areprocessed to prepare a composite preparation or the mannuronate oligosaccharides and the traditional antibacterial agent are respectively processed to prepare individual preparations which can be used simultaneously or sequentially. The preparation method has the advantages of simplicity, easiness in control, and obtaining of the mannuronate oligosaccharides having the polymerization degree focusing on 2-6 through degradation under specific conditions and purifying treatment. The mannuronate oligosaccharides can be combined with the traditional antibacterial agent to significantly weaken theresistance of common clinic drug-resistant bacteria to corresponding antibacterial agents, and are suitable for large-scale development applications.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a preparation method of mannuronic acid oligosaccharide and its application in the preparation of a combined antibacterial agent for inhibiting drug-resistant bacteria. Background technique [0002] With the widespread, excessive and unreasonable use of antibacterial agents, especially quinolones and cephalosporins, drug-resistant bacteria continue to increase and spread, causing adverse reactions and double infections. Increased morbidity and mortality have caused great harm to human health. The treatment of infections caused by drug-resistant bacteria is facing severe clinical challenges. The drug-resistant mechanism of drug-resistant bacteria is complex, and the treatment effect of traditional antibacterial drugs is becoming less and less ideal. At present, there is an urgent need for new antibacterial drugs or combined antibacterial agents that are different from the tradit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H3/06C07H3/04C07H1/00C08B37/00A61K31/546A61K31/702A61K31/715A61P31/04
CPCA61P31/04A61K31/546A61K31/702A61K31/715C07H1/00C07H3/04C07H3/06C08B37/0033
Inventor 郭怀忠崔秀彦曹丽军卢凯张利沙
Owner HEBEI UNIVERSITY
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