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Beta-lactan antibiotic grafted polymer and application thereof

A technology of grafting polymers and lactams, applied in the field of biochemical analysis, can solve the problems of heavy workload, long time, inability to generate new lactamase genes and mutant genes, etc., and achieve the effect of reducing affinity

Active Publication Date: 2018-01-12
吉林省爱诺德生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The advantage of this method is that it is quantitative and accurate, but its main disadvantage is that the operation is complicated and time-consuming.
In recent years, the drug resistance detection method of β-lactam antibiotics based on antibiotic inactivation enzyme gene or biochemical activity has become more and more popular. The advantage of this method is that it can quickly determine the expression of specific β-lactamase from the molecular level. The disadvantage is that the primers used can only be used to detect the known genes of β-lactam antibiotics, and cannot be used for the detection of new lactamase genes and mutant genes. β-lactam antibiotics
The experimental method of directly detecting the bacterial lactamase activity of β-lactam antibiotics has also been reported. For example, Bernabeu, Pirelli et al. used ultraviolet spectrophotometry to analyze the level of β-lactam antibiotics hydrolyzed by bacterial lysates to provide information on their drug resistance. But this method is labor-intensive and still takes hours to produce test results

Method used

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  • Beta-lactan antibiotic grafted polymer and application thereof
  • Beta-lactan antibiotic grafted polymer and application thereof
  • Beta-lactan antibiotic grafted polymer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Preparation of Cefepime Grafted Polymer

[0024] Step 1: Add 100ml of dichloroethane into the three-necked flask, add 10g of hydroxymethyl polystyrene resin and 20g of bisglycidyl ether into the reactor and mix evenly, slowly add 1ml of 40% sodium hydroxide aqueous solution dropwise at room temperature, After reacting for 5 hours, filter, and the filter cake is fully washed with 200ml ethylene dichloride to obtain a grafted polymer;

[0025] Step 2: Disperse the grafted polymer obtained by the product of step 1) in 100ml THF solution, add 20g cefepime into the reactor to carry out ring-opening reaction, react for 8 hours, filter the reaction solution, and filter the filter cake fully with 200ml THF Washing to finally obtain the cefepime grafted polymer;

[0026] Step 3: Suspend the cefepime grafted polymer obtained in step 2) in 50mM Tris, PBS of pH 7.0, adopt a wet packing method, and fill 1 ml of gel into each column. After the column is installed, use 5ml , 0.01%Na...

Embodiment 2

[0028] Preparation of Ceftaroline Grafted Polymer

[0029] Step 1: Add 100ml tetrahydrofuran into the three-necked flask, add 10g hydroxymethyl polystyrene resin and 20g bisglycidyl ether into the reactor and mix evenly, slowly add 0.3g SDS at room temperature, react for 5 hours, filter, filter cake Fully wash with 200ml tetrahydrofuran to obtain graft polymer;

[0030] Step 2: Disperse the graft polymer obtained by the product of step 1) in 100ml DMF solution, add 20g cefepime into the reactor to carry out ring-opening reaction, react for 8 hours, filter the reaction solution, and wash the filter cake fully with 200ml DMF , to finally obtain the cefepime grafted polymer;

[0031] Step 3: Suspend the cefepime grafted polymer obtained in step 2) in 50mM Tris, PBS of pH 7.0, adopt a wet packing method, and fill 1 ml of gel into each column. After the column is installed, use 5ml , 0.01%NaN 3 -PBS sterile buffer continues to pass through the column. When the top of the column ...

Embodiment 3

[0033] Preparation of Cefditoren Axetil Graft Polymer

[0034] Step 1: Add 100ml of DMF to the three-necked flask, add 10g of hydroxymethyl polystyrene resin and 20g of bisglycidyl ether into the reactor and mix evenly, slowly add 0.4g of 16-alkyl sodium sulfate at room temperature, and react for 5 hours , filter, and the filter cake is fully washed with 200ml DMF to obtain a grafted polymer;

[0035] Step 2: Disperse the graft polymer obtained by the product of step 1) in 100ml of dichloromethane solution, add 20g of cefepime into the reactor to carry out ring-opening reaction, react for 8 hours, filter the reaction solution, and use 200ml Dichloromethane fully washes, finally obtains cefditoren axetil graft polymer;

[0036] Step 3: The cefditoren axetil graft polymer obtained in step 2) was suspended in 50 mM Tris, pH 7.0 PBS. Wet column packing is adopted, each column is loaded with 1 ml gel, after the column is installed, use 5ml, 0.01% NaN 3 -PBS sterile buffer contin...

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Abstract

The invention provides a beta-lactan antibiotic grafted polymer and an application thereof. A method comprises the following steps: subjecting hydroxylmethyl polystyrene resin and diglycidyl ether tocondensation in the presence of a catalyst; subjecting an obtained product to a ring-opening reaction with amino on a 4-bit side chain of beta-lactan antibiotics, and carrying out repeated washing, soas to obtain the grafted polymer for detecting drug resistance of beta-lactan antibiotics; packing the polymer to a column, and monitoring peak appearance time of a sample in effluent by using an ultraviolet detector; and carrying out bromocresol purple detection on the packed column, thereby rapidly obtaining pharmaceutical sensitive information on the sample. The method is applicable to proteinsamples and is also applicable to the detection on the drug resistance of the beta-lactan antibiotics through bacterial living bodies and a thallus lysate, so that the beta-lactan antibiotic graftedpolymer can be applied to rapid drug resistance analysis in clinical laboratories and used for instructing doctors to determine clinical medication schemes as soon as possible.

Description

technical field [0001] The invention relates to a β-lactam antibiotic graft polymer and its application. It belongs to the field of biochemical analysis. Background technique [0002] Alterations in the penicillin-binding protein (PBP) often lead to two clinically important resistance phenotypes: reduced affinity for β-lactam antibiotics, resulting in bacterial resistance to β-lactam antibiotics; acquired resistance is caused by Plasmid-mediated, after the bacteria acquire the drug-resistant gene, they produce a large amount of β-lactamase (instead of PBPs), which slowly inactivates the resistant enzyme penicillin, showing drug resistance, mostly borderline drug resistance; traditional antibiotic sensitivity The method for detecting β-lactam antibiotics is the agar dilution method, that is, the minimum inhibitory concentration MIC method for detecting β-lactam antibiotics. The advantage of this method is that it is quantitative and accurate, but its main disadvantage is th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F212/14C08F8/32C08F8/08G01N30/02G01N30/56
Inventor 许文革宋立明杨琨
Owner 吉林省爱诺德生物工程有限公司
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