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Application and method of killing drug-resistant bacteria with blebbistatin analogs activated by blue light

A technology of drug-resistant bacteria and analogs, applied in the field of biomedicine, can solve the problems of no biological target, no biological or medical application, no biological activity, etc., to achieve good sterilization effect, solid and strong technical support, and sterilization efficiency. improved effect

Active Publication Date: 2021-11-23
THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 and method of killing drug-resistant bacteria with blebbistatin analogs activated by blue light
  • Application and method of killing drug-resistant bacteria with blebbistatin analogs activated by blue light
  • Application and method of killing drug-resistant bacteria with blebbistatin analogs activated by blue light

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 present invention relates to the application and method of killing drug-resistant bacteria with blebbistatin analogues activated by blue light, wherein the blebbistatin analogues include (R)-blebbistatin; the drug-resistant bacteria include Staphylococcus aureus, Escherichia coli, Enterococcus, Pseudomonas Monas, Acinetobacter baumannii; the wavelength of the blue light is 420nm. In the present invention, (R)-blebbistatin, which has no mammalian target and no biological activity, is activated by blue light with a wavelength of 420nm and used to kill drug-resistant bacteria such as Staphylococcus aureus, Escherichia coli, Enterococcus, Pseudomonas aeruginosa, Acinetobacter baumannii has a significant bactericidal effect. The present invention also finds that it is safe to use (R)-blebbistatin molecules for 60 minutes of treatment in the living body; and the bactericidal efficiency increases with the increase of the light time, and there is no risk of inducing drug resistance. The invention is not limited by bacterial drug resistance, and provides solid and powerful technical support for the treatment of clinical multi-drug 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 Patents(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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