Application of blue light activated blebbistatin analogue to killing of drug-resistant bacteria and method

A technology of drug-resistant bacteria and analogs, applied in the field of biomedicine, can solve the problems of no biological targets, no biological activity, no research on biological or medical applications, etc., achieve good bactericidal effect, improve bactericidal efficiency, solid and powerful technology The effect of support

Active Publication Date: 2020-02-21
THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aaron F.Straight et al. (Aaron F.Straight, Dissecting Temporal and Spatial Control of Cytokinesis with a Myosin II Inhibitor, Science, 2003) disclosed left-handed blebbistatin ((S)-blebbistatin) and right-handed blebbistatin ((R)-blebbistatin ), but the dextrorotatory blebbistatin has been shown to have no biological targets and no biological activity in mammalian cells
No biological or medical applications of (R)-blebbistatin have been studied so far

Method used

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  • Application of blue light activated blebbistatin analogue to killing of drug-resistant bacteria and method
  • Application of blue light activated blebbistatin analogue to killing of drug-resistant bacteria and method
  • Application of blue light activated blebbistatin analogue to killing of drug-resistant bacteria and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method (concentration gradient sterilization) for killing clinical drug-resistant Acinetobacter baumannii GD0302 with blue light-activated blebbistatin analog (R)-blebbistatin, comprising the steps of:

[0041] 1. Streak culture of clinical drug-resistant Acinetobacter baumannii on a culture plate to obtain monoclonal bacteria.

[0042] 1.1) Take out the clinical drug-resistant Acinetobacter baumannii (Code: GD0302) stored at -80°C, dissolve it on ice, dip 1 μL of it with a ring inoculation stick and streak it on the LB agar plate several times.

[0043] 1.2) The upside-down plate was cultured in an incubator for 12 hours to obtain monoclonal bacteria.

[0044] 2. Inoculate into the culture medium to make the clinical drug-resistant Acinetobacter baumannii reach the logarithmic phase.

[0045] 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.

[0046] 2.2) Take 1 / 10 of the overnight ba...

Embodiment 2

[0061] A method (density gradient sterilization) for killing clinical drug-resistant Acinetobacter baumannii GD0302 with blue light-activated blebbistatin analog (R)-blebbistatin, comprising the steps of:

[0062] The culture material, apparatus and process of this embodiment are basically the same as embodiment 1, the difference is:

[0063] (1) being different from 3.1 in embodiment 1), the working density of bacteria becomes 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 8 cfu / mL, 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.

[0064] (2) Analysis of results

[0065] This embodiment adopts drug-resistant Acinetobacter baumannii strain (number: GD0302), and the density is 10 8 cfu / mL,10 7 cfu / mL,10 6 cfu / mL,10 5 cfu / mL,10 4 cfu / mL,10 3 cfu / mL, mixed with 6 μM (R)-blebbistatin and then illuminated at 420 nm for 60 min. It was observed that (R)-blebbis...

Embodiment 3

[0067] A method for killing clinical drug-resistant Acinetobacter baumannii GD0302 with blue light-activated blebbistatin analogue (R)-blebbistatin (sterilization at different blue light wavelengths), comprising the steps of:

[0068] The culture material, apparatus and process of this embodiment are basically the same as embodiment 1, the difference is:

[0069] (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.

[0070] (2) Different from 3.2) in Example 1, the concentration of (R)-blebbistatin used in Example 3 is 0 μM, 1 μM, 6 μM, that is, the 2× working concentration is: 0 μM, 2×1 μM, 2×6 μM.

[0071] (3) Different from 3.5) in Example 1, Example 3 uses 3 kinds of blue light sources with different wavelengths, namely 360nm, 420nm and 460nm blue light sources.

[0072] (4) Different from 4.1) in Example 1, according to the sterili...

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Abstract

The invention relates to an application of a blue light activated blebbistatin analogue to killing of drug-resistant bacteria and a method. The blebbistatin analogue comprises (R)-blebbistatin; the drug-resistant bacteria comprise staphylococcus aureus, escherichia coli, enterococcus, pseudomonas aeruginosa and acinetobacter baumannii; and blue light wavelength is 420 nm. (R)-blebbistatin withoutmammalian targets or bioactivity is utilized for killing the drug-resistant bacteria such as the staphylococcus aureus, the escherichia coli, the enterococcus, the pseudomonas aeruginosa and the acinetobacter baumannii after being activated by blue light with wavelength of 420 nm and has a remarkable bactericidal effect. According to the invention, it is safe to adopt (R)-blebbistatin molecules ina living body for 60 min treatment; and besides, the bactericidal efficiency is improved with increase of illumination time, and the risk of induced resistance is eliminated. The application and themethod are not limited by bacterial drug resistance, and solid and powerful technical support is provided for clinical treatment of multidrug-resistant bacteria.

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 bacteria with blue light-activated blebbistatin analogues. Background technique [0002] Common pathogenic bacteria in hospital-acquired infections (ie, nosocomial infections) include gram-positive bacteria (eg, Staphylococcus aureus, Enterococcus) and gram-negative bacteria (eg, Pseudomonas Acinetobacter, Escherichia coli). Staphylococcus aureus is a common food-borne pathogenic microorganism, which often parasitizes in the skin, nasal cavity, throat, intestines and stomach, and purulent sores, and can easily cause food poisoning. Enterococcus widely exists in the human digestive tract and is an important pathogenic bacteria of nosocomial infection, which can cause urinary tract infection, skin and soft tissue infection, abdominal infection, meningitis and so on. However, due to the inherent drug resistance of enteroco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61P31/04A01N43/90A01P1/00A61L2/00
CPCA01N43/90A61K41/0057A61L2/0052A61P31/04A61K41/0038A01N25/00
Inventor 黄乃淇谢永丽周子原袁静李明德高磊张明霞刘映霞刘磊
Owner THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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