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Application of methysticin

A kind of technology of anesthetizing capsaicin and application, applied in the field of medicine

Active Publication Date: 2013-12-11
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Anesthesia capsaicin Methysticin was purchased from Shanghai Tongtian Biotechnology Co., Ltd. ( http: / / tautochem.labscn.com / ), so far the drug has been reported to inhibit human liver microsomal cytochrome P450 ( Mathews JM , et al, Drug Metab Dispos. 2005Oct; 33(10): 1555-63.) But there is no report about the drug as a VicK inhibitor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: In vitro kinase activity test of traditional Chinese medicine monomers

[0020] In vitro kinase activity assay application provided by Promega Plus Luminescent Kinase Assay kit, the experimental steps are strictly in accordance with the instructions attached to the kit. Proceed as follows:

[0021] 1. Main reagents and instruments:

[0022] ATP was purchased from Sigma;

[0023] Plus Luminescent Kinase Assay was purchased from Promega Company Cat#V3772;

[0024] Enzyme reaction buffer: 40mM Tris-HCl[pH7.5], 20mM MgCl 2 , 0.1mg / ml BSA;

[0025] The microplate reader is Tecan GENios; the microplate plate is costar R solid white, flat-bottom96-well plates; electric heating constant temperature water bath box: Beijing Changan Scientific Instrument Factory HH-W21CY600 type.

[0026] 2. Experimental steps:

[0027] Detect the inhibitory effect of the traditional Chinese medicine monomer concentration of 200 μM on VicK kinase:

[0028] 100% dimethyl sul...

Embodiment 2

[0042] The mensuration of embodiment 2 minimum inhibitory concentration (MIC, minimal inhibitory concentration) and minimum bactericidal concentration (MBC, minimal bactericidal concentration)

[0043] 1. MIC adopts micro broth dilution method

[0044] 1) Aseptic operation, add antimicrobial drug solutions of different concentrations after doubling dilution to a sterilized 96-well polystyrene plate, add drug solution to the 1st to 11th wells, 10 μl per well, and do not add to the 12th well Drugs were used as growth control, and DMSO control was also done at the same time.

[0045] 2) Dilute the bacterial suspension prepared by the growth method or the direct bacterial suspension method with a concentration equivalent to 0.5 McFarland turbidimetric standard to 1:1000 in broth, add 200 μl to each well, seal it and place it under normal conditions at 37°C. In the air incubator, incubate for 16-20h to judge the result.

[0046] 3) Judging the results, the minimum drug concentrat...

Embodiment 3

[0052] Example 3: Anesthesia capsaicin enhances the sensitivity of bacteria to penicillin

[0053] Steps: 1. Add 200 μL of clinical Streptococcus pneumoniae and Staphylococcus aureus (from Children’s Hospital) to 5ML C+Y medium to grow to OD600 of 0.5, dilute 10 times to a concentration of 107 CFU / ML

[0054]2. Add 25 μL of C+Y medium to each well of the 96-well plate, add PNC (penicillin) at a concentration lower than the MIC of each bacterium in the first well, serially dilute, and then add 25 μL of the combination drug. At the same time, no drug was added as a positive control and 5% DMSO was added as a control, and a parallel sample was made for each test.

[0055] 3. Add 150 μL of bacterial solution to each well and incubate overnight at 37 degrees, so that the minimum drug concentration that completely inhibits bacterial growth in the small well is MIC

[0056] 4. Plate the wells in which no bacteria grow in each group of drugs in turn, and the number of colonies on the...

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Abstract

The invention discloses the application of methysticin used as a histidine kinase VicK inhibitor in vitro or application of methysticin in preparation of the histidine kinase VicK inhibitor. The methysticin has a certain antibacterial effect on in-vivo streptococcus pneumoniae and clinical penicillin resistant streptococcus pneumoniae. Moreover, the medicament has a good antibacterial effect on streptococcus pneumoniae after being combined with penicillin, and has obvious bacteriostasis on staphylococcus aureus.

Description

technical field [0001] This research involves the field of medical technology, specifically the application of caperin, an anesthetic monomer of traditional Chinese medicine, as an inhibitor of histidine kinase VicK. Background technique [0002] The two-component system (TCS) is the main signal transduction system in bacteria. It not only participates in the basic life activities of bacteria, but more importantly, it is closely related to the pathogenicity and drug resistance of many pathogenic bacteria. Bacterial TCS systems have the following two characteristics: Histidine kinases and response regulators in different types of bacteria have obvious homology, especially the amino acid residues in the active center, so this system regulation method is common in bacteria sex. The TCS regulatory system is unique to bacteria (mainly Gram-positive bacteria and a few Gram-negative bacteria), and mammals do not have this signal transduction mechanism. Therefore, histidine kinase...

Claims

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Application Information

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IPC IPC(8): A61K31/366A61K31/43A61P31/04
Inventor 傅若秋卢来春李卓恒于彩平胡大强
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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