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Method for sieving active component or matter and active component therefrom

A technology of active components and combined activities, applied in biochemical equipment and methods, medical preparations containing active ingredients, measurement/inspection of microorganisms, etc., can solve the problems of penicillin resistance, wide application restrictions, and anti-MRSA infection effective substances The basics are not clear, etc.

Inactive Publication Date: 2007-11-14
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the late 1930s, sulfa drugs were first used to treat Staphylococcus aureus infectious diseases, but their widespread use was limited due to the emergence of bacterial resistance
After the advent of penicillin in the early 1940s, the infectious diseases caused by Staphylococcus aureus were well controlled, but with the widespread use of penicillin, some Staphylococcus aureus produced penicillinase to hydrolyze the β-lactam ring, showing that they were resistant to penicillin drug resistance
At the same time, clinical and experimental studies have shown that some natural medicine preparations can be taken orally or externally to treat MRSA infection. However, due to the complex composition of natural medicine, especially limited by methodology, the effective material basis for anti-MRSA infection in natural medicine is still unclear.

Method used

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  • Method for sieving active component or matter and active component therefrom
  • Method for sieving active component or matter and active component therefrom
  • Method for sieving active component or matter and active component therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1 : Coating of the sample cell

[0101] 1.1 Experimental method:

[0102]Biotinylation of PBP2a: Strictly follow the instructions of the Sigma kit: dissolve 1 tube of BAC-Sulfo NHS with 30ul DMSO, add 1ml of 0.01M PBS, immediately remove 10ul and add to 0.2ml (8mg / ml) PBP2a; Shake for 30 minutes, after treating the micro chromatography column, add the above-mentioned PBP2a, centrifuge at 700gx2min, and collect the solution at the bottom of the tube, which is the purified biotin-labeled PBP2a;

[0103] According to the coating process of the biotin plate sample pool of the affinity-type specific and selective biosensor IAsys plus (Affinity-sensorIAsys plus) of Thermo Company, PBP2a was coated in the sample pool, and the amount of coated PBP2a was calculated. The specific operation is as follows : ①The coating temperature of the instrument is set at 20°C to 25°C, the stirring rate is set at 85 times / s, and the data collection is 1 time / s; ②Put it into the sample...

Embodiment 2

[0105] Example 2 : Determination results of the binding reaction between 115 kinds of natural medicine extracts and PBP2a

[0106] 2.1 Experimental method: 115 kinds of natural medicine extracts were prepared, 5 μl of them were sampled respectively, and the binding reaction with PBP2a was measured, and the detection method was the same as above.

[0107] 2.2 Experimental results: the binding value of water extracts of 25 kinds of traditional Chinese medicines such as Maodongqing, Golden Buckwheat, Tripterygium wilfordii and PBP2a is greater than 100RU. The results in Figures 2A and 2B show that there are large differences in the binding of natural medicines to PBP2a (RU: 0 arc second to 281 arc second), indicating that there may be large differences in the substances that these natural medicines can bind to PBP2a.

Embodiment 3

[0108] Example 3 : Depletion Experiment of Traditional Chinese Medicine and Quantitative PBP2a

[0109] 3.1 Crush the 25 traditional Chinese medicines with high affinity and accurately weigh 2g and divide them into two groups of test tubes and number them. Add 10ml of distilled water to the first group in a water bath at 100°C for 1 hour. After cooling, centrifuge at 5000r / min for 3 minutes to collect the supernatant. For standby, the final crude drug concentration of the sample is 1g·ml -1 ; Take 100 μl of the water extract sample, divide it into two groups 60 Co-irradiated EP tubes, add an equal volume of 50ngml to the first group -1 The PBP2a in the sample was used to consume part of the active substances in the sample. The second group added an equal volume of normal saline as the control group. After the two groups of samples were fully mixed, they were bathed in water at 37°C for 30 minutes, and after cooling, they were centrifuged at 4000rpm for 5 minutes. The supern...

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Abstract

One kind of union biosensor technology and isolation technique, From natural medicine, selection and separation have the combination PBP2a active method of organizing minute or active material, the method will PBP2a coating on the sample pool biosensor match as a solid base to extract natural medicines the mobile phase of the selected combination with PBP2a in the role of natural medicine, will be an integrated component of the higher value after isolation and purification, has been a collaborative antagonistic MRSA activity combined with PBP2a occurred role of the active component, the Preparation synergistic components can be used to beta - lactam antibiotics treatment of MRSA infection drugs or lead compounds. This method has time-saving, the high flux, to reduce effort, the specificity to be strong, the result accurate, can real-time monitoring many kinds of merits.

Description

technical field [0001] The invention relates to a method for screening and isolating active components or substances and the resulting active components, in particular to a combination of biosensor technology and separation technology for screening and isolating active components or substances that bind to PBP2a from natural medicines. The method for the active substance, and the active component capable of combining with PBP2a obtained by screening and separating natural medicines by using the method. Background technique [0002] Research experience at home and abroad shows that it is very promising to obtain new drugs for treating intractable diseases from natural lead compounds, and the hit rate of effective monomers screened from natural compounds is higher than that of synthetic compounds. Clinical and experimental studies have shown that some natural medicine preparations can exert therapeutic effects on pathogenic molecules in symptomatic diseases through various mec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18C12Q1/25A61K36/718A61P31/04
CPCY02A50/30
Inventor 周红和生琦董燕郑江
Owner ARMY MEDICAL UNIV
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