Application of blue light activated (S)-blebbistatin molecules to killing of drug-resistant acinetobacter baumannii, and method for killing drug-resistant acinetobacter baumannii through blue light activated (S)-blebbistatin molecules

An Acinetobacter baumannii, drug-resistant technology, applied in the field of biomedicine, can solve the problems of difficult drug administration, ineffective mammalian cytotoxicity, and poor efficiency, and achieve the effect of solid and powerful technical support

Active Publication Date: 2020-02-07
THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Li (M.D.Li, N-K Wong, et al.J.Am.Chem.Soc.140(46):15957-15968(2018)) disclosed that (S)-blebbistatin molecules can produce highly oxidative Hydroxyl radical ( · OH), but Li et al. previously used human and mouse skin and stomach tumor cell models, and the dose of tumor cells used was 10 to 25 μM (25 μM is the upper limit of (S)-blebbistatin water solubility), and its efficiency was not good. (S)-blebbistatin has no obvious toxic effect on mammalian cells, which affects its anti-tumor application
There are many ways to generate hydroxyl radicals in chemical systems, such as Fenton reaction and ultraviolet irradiation, but they have problems such as difficulty in administration or low safety, and are not suitable for anti-infection treatment

Method used

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  • Application of blue light activated (S)-blebbistatin molecules to killing of drug-resistant acinetobacter baumannii, and method for killing drug-resistant acinetobacter baumannii through blue light activated (S)-blebbistatin molecules
  • Application of blue light activated (S)-blebbistatin molecules to killing of drug-resistant acinetobacter baumannii, and method for killing drug-resistant acinetobacter baumannii through blue light activated (S)-blebbistatin molecules
  • Application of blue light activated (S)-blebbistatin molecules to killing of drug-resistant acinetobacter baumannii, and method for killing drug-resistant acinetobacter baumannii through blue light activated (S)-blebbistatin molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The application and method of blue light activation (S)-blebbistatin molecule killing drug-resistant Acinetobacter baumannii (concentration gradient sterilization), including the following steps:

[0029] 1. Streak culture of clinical drug-resistant Acinetobacter baumannii on a culture plate to obtain monoclonal bacteria.

[0030] 1.1) Take out the clinical drug-resistant Acinetobacter baumannii (number: GD0302) stored at -80°C (see figure 2 ), after dissolving on ice, use a circular inoculation stick to dip 1 μL and streak it on the LB agar plate several times.

[0031] 1.2) The upside-down plate was cultured in an incubator for 12 hours to obtain monoclonal bacteria.

[0032] 2. Inoculate into the culture medium to make the clinical drug-resistant Acinetobacter baumannii reach the logarithmic phase.

[0033] 2.1) Pick 1-4 monoclonal bacteria and culture them overnight on a shaker at 220 rpm in a shaker tube filled with 3 mL of LB medium.

[0034] 2.2) Take 1 / 10 of...

Embodiment 2

[0058] Application and method of killing drug-resistant Acinetobacter baumannii by activating (S)-blebbistatin molecules with blue light (density gradient sterilization)

[0059] The culture material, apparatus and process of this embodiment are basically the same as embodiment 1, the difference is:

[0060] (1) being different from 3.1 in embodiment 1), embodiment 2 has improved the upper limit of bacterial density and relaxed lower line density on the basis of embodiment 1, i.e. 10 8 cfu / mL-10 3 cfu / mL. Bacteria were serially diluted with 10-fold density gradient in PBS to 2×working density, that is, 2×10 7 cfu / mL, 2×10 6 cfu / mL, 2×10 5 cfu / mL, 2×10 4 cfu / mL, 2×10 3 cfu / mL.

[0061] (2) Analysis of results

[0062] This embodiment adopts drug-resistant Acinetobacter baumannii strain (number: GD0302), and the density is 10 7 cfu / mL,10 6 cfu / mL,10 5 cfu / mL,10 4 cfu / mL,10 3 cfu / mL, after mixing with 2μM and 10μM (S)-blebbistatin, light at 420nm for 60min. It can b...

Embodiment 3

[0064] Application and method of killing drug-resistant Acinetobacter baumannii by activating (S)-blebbistatin molecules with blue light (different blue light wavelengths)

[0065] The culture material, apparatus and process of this embodiment are basically the same as embodiment 1, the difference is:

[0066] (1) Different from 3.1) in Example 1, the density of drug-resistant Acinetobacter baumannii used in Example 3 is 10 8 cfu / mL, that is converted to 2 times the working density is 2×10 8 cfu / mL.

[0067] (2) Different from 3.2) in Example 1, the concentration of (S)-blebbistatin used in Example 3 is 0 μM and 2 μM, that is, the 2× working concentration is 0 μM and 2×2 μM.

[0068] (3) Different from 3.5) in Example 1, Example 3 uses two kinds of blue light sources with different wavelengths, namely 420nm and 460nm blue light sources.

[0069] (4) Different from 4.2) in Example 1, the dilution factor used in Example 3 was 20,000 times, and 100 μL was evenly spread on the ...

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Abstract

The invention relates to application of blue light activated (S)-blebbistatin molecules to killing of drug-resistant acinetobacter baumannii, and a method for killing the drug-resistant acinetobacterbaumannii through the blue light activated (S)-blebbistatin molecules. (S)-blebbistatin molecules are used for cooperating with blue light, and the (S)-blebbistatin molecules generate strong-oxidativehydroxyl radicals (.OH) under blue light irradiation, and can kill the drug-resistant acinetobacter baumannii. (S)-blebbistatin and a bacterial solution are mixed evenly to be irradiated under the blue light and then smeared or dotted on the surface of a flat plate for a period of time, and then it can be observed that the clinical drug-resistant acinetobacter baumannii is killed. The sterilizingefficiency is improved with increasing of the time, and the inducible drug resistance risk is avoided under the condition without light. The application and the method are not limited by bacterial drug resistance, and provide solid and powerful technical support for treatment of the clinical multiple-drug-resistant acinetobacter baumannii.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application and method of killing drug-resistant Acinetobacter baumannii by activating (S)-blebbistatin molecules with blue light. Background technique [0002] Acinetobacter baumannii is a non-fermenting Gram-negative bacterium that widely exists in the natural environment, human skin, digestive system, and urinary system. Acinetobacter baumannii mainly targets trauma infections, such as ventilator-associated pneumonia. The drug-resistant phenotype of Acinetobacter baumannii was prominent, especially for carbapenem antibiotics and quinolone antibiotics. With the emergence of multidrug-resistant Acinetobacter baumannii, tigecycline and polymyxin antibiotics have become the last line of defense, but these two types of drugs also have problems such as strong side effects and poor pharmacokinetics. At present, due to the serious lack of research and development of new an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61P31/04C12Q1/04C12R1/01
CPCA61K41/0057A61P31/04C12Q1/04G01N2333/212
Inventor 黄乃淇李明德谢永丽袁静周子原高磊张明霞刘映霞刘磊
Owner THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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