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Method for enhancing antibiotic antibacterial sensitivity by NO and application

An antibiotic and sensitivity technology, applied in the field of biomedicine, can solve the problems of difficult pathogenic bacteria and complex drug resistance mechanism, and achieve the effects of enhancing sensitivity, avoiding the generation of drug resistance, and reducing the amount of use

Pending Publication Date: 2022-03-18
MICROBIOLOGY INST OF SHAANXI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the complex mechanism of PA drug resistance and the outstanding drug resistance of many clinical antibiotics, it is difficult for the commonly used antibiotics to effectively kill pathogenic bacteria at low doses. Therefore, it is urgent to develop a simple and effective method to improve the efficacy of antibiotics and their antimicrobial susceptibility

Method used

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  • Method for enhancing antibiotic antibacterial sensitivity by NO and application
  • Method for enhancing antibiotic antibacterial sensitivity by NO and application
  • Method for enhancing antibiotic antibacterial sensitivity by NO and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The effect test of exogenous NO donor SNP combined with aminoglycoside and β-lactam antibiotics on the growth characteristics of Pseudomonas aeruginosa adherent bacteria (Sessilebacteria, that is, bacteria adhered to the surface of the medium) is as follows:

[0041] (1) Antibiotic susceptibility test of PA bacteria of adhesion type

[0042] The preserved strain PAO1 was inoculated in M9 medium and cultured at 37°C until OD 600nm is 0.5. This is PAO1 liquid seed solution. Add slides to the 6-well plate, add M9 medium to the wells, and then add seed solution to each well at an inoculation amount of 0.5%. The bacterial solution in the wells was pretreated by adding NO donors with different concentrations of 1-100 μM SNP at 37°C for 24 hours, and the blank was set as the control well. The slides were taken out, and the slides were gently washed with PBS 3 times to remove planktonic bacteria. Put the cleaned slides back into a new 6-well plate and add M9 medium and antib...

Embodiment 2

[0049] Exogenous NO donor SNP enhances the susceptibility test of planktonic and sessile bacteria to aminoglycosides and β-lactam antibiotics of Pseudomonas aeruginosa as follows:

[0050] (1) Test method

[0051] After cultured with SNP pretreatment on the round cell culture dish for 24 hours, a group without SNP pretreatment was cultured at the same time as a control. For the specific process, see the attached Figure 6 As indicated, antimicrobial susceptibility of planktonic and adherent cells (settle cells) was analyzed by two methods.

[0052] Antibiotic susceptibility analysis of planktonic bacteria: the supernatant on the circular cell culture dish was diluted 10 times into the M9 medium containing 5 μg / ml tobramycin, and then cultured at 37°C for 72 hours. Measure OD every 2h 600nm To assess the effect of NO on the susceptibility of bacterioplankton.

[0053] Antibiotic susceptibility analysis of sessile bacteria: Treat the adherent PA cells on the glass slide at th...

Embodiment 3

[0061] The effect of endogenous NO on alginate synthesis in Pseudomonas aeruginosa was tested as follows:

[0062] In this experiment, PAO1 is the PA model strain, Δnor is the nitric oxide reductase deletion strain, and Δnor+ is the nitric oxide reductase replenishment strain.

[0063] The function of NO reductase Nor is to convert intracellular NO into N 2 O, to avoid excessive accumulation of intracellular NO. Therefore, the Δnor mutant will continue to accumulate endogenous NO to produce some biological effects. In this experiment, the influence of endogenous NO on PA sensitivity was determined by culturing Δnor mutant strains and nitric oxide reductase replenishing strains, and comparing alginate levels in PAO1, Δnor and Δnor+ strains.

[0064] Determination of alginate levels in PAO1, Δnor and Δnor+ strains, see attached Figure 12 . attached by Figure 12 The data shows that, compared with the PAO1 strain, the alginate levels in the Δnor strain and the Δnor+ strain ...

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Abstract

The invention discloses a method for enhancing antibacterial sensitivity of pseudomonas aeruginosa to antibiotics by NO. NO is provided by an exogenous NO donor, is provided by an endogenous NO donor or is accumulated by means of endogenous NO metabolism regulation, so that the antibacterial sensitivity of pseudomonas aeruginosa to one or more combined antibiotics in aminoglycoside or beta-lactam antibiotics can be enhanced, and the antibacterial sensitivity of pseudomonas aeruginosa to antibiotics can be enhanced. Meanwhile, an exogenous NO donor is provided and comprises compounds, such as SNP and GSNO, capable of directly or indirectly providing NO to an infected part, an endogenous NO donor comprises drugs, such as nitroglycerin, capable of generating NO through human metabolism, and regulation substances for endogenous NO metabolism comprise an inhibitor for inhibiting the NO metabolic process of the pseudomonas aeruginosa and a reinforcing agent for host NO synthetase. The invention finds a novel method capable of effectively enhancing the anti-sensitivity of the pseudomonas aeruginosa to the antibiotics, and related antibacterial sensitive drugs can be applied to preparation of antibacterial drugs for pseudomonas aeruginosa infection.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method and application for enhancing the antibacterial susceptibility of Pseudomonas aeruginosa to antibiotics by NO. Background technique [0002] Aminoglycoside antibiotics (Aminoglycosides, AGs) have a very broad antibacterial spectrum, mainly including streptomycin, tobramycin, gentamicin, amikacin, etc., their broad-spectrum activity, rapid bactericidal effect and good chemical properties And pharmacokinetic properties, making it clinically applicable to a variety of infection types of drugs, in addition to broad-spectrum killing of Gram-negative bacteria and positive bacteria, it is often combined with β-lactam antibiotics Application enhances the drug effect; β-lactam antibacterial drugs mainly include penicillins, cephalosporins and carbapenems, which mainly exert an inhibitory effect on the cell wall mucopeptide synthase of pathogenic bacteria, causing ...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K33/26A61P31/04
CPCA61K45/06A61K33/26A61P31/04A61K2300/00
Inventor 万一陈畅高磊乔新华王琰
Owner MICROBIOLOGY INST OF SHAANXI
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