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Anti-infective pharmaceutical use of hordenine or its combined antibiotics

A technology of hordenine and antibiotics, applied in the field of medicine, to achieve the effect of inhibiting bacterial infection

Active Publication Date: 2020-05-08
HAINAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no relevant report about the anti-infective research of hordenine

Method used

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  • Anti-infective pharmaceutical use of hordenine or its combined antibiotics
  • Anti-infective pharmaceutical use of hordenine or its combined antibiotics
  • Anti-infective pharmaceutical use of hordenine or its combined antibiotics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Synthesis of signal molecules regulated by quorum sensing: inoculate a single colony of Pseudomonas aeruginosa PAO1 into NB medium, and culture overnight at 37°C and 150rmp. Tune OD 620 = 0.5, take 50 μL of the OD-adjusted bacterial solution and add it to 50 mL NB medium at a ratio of 1:1000, and add different concentrations of hordenine at the same time, and incubate at 37° C. and 200 rpm for 48 hours. Centrifuge at 12,000rpm at 4°C for 10min, extract the supernatant with the same volume of acidified ethyl acetate (containing 0.5% formic acid) three times, evaporate to dryness at 35°C under reduced pressure, dissolve the dry matter in methanol, and measure the signal by LC / MS / MS molecular content. Determine the type of signal molecule according to the ion fragment peak and retention time, and determine the relative content according to the peak area. Test results such as figure 1 , where A(a) is the MS / MS mass spectrum of the signal molecule C4-HSL, A(b) is the MS / M...

Embodiment 2

[0028] Effects on the expression of virulence factors regulated by quorum sensing: inoculate a single colony of Pseudomonas aeruginosa PAO1 into NB medium, and culture overnight at 37°C and 150rmp. Tune OD 620 = 0.5, take 50 μL of the OD-adjusted bacterial solution and add it to 50 mL NB medium at a ratio of 1:1000, and add different concentrations of hordenine at the same time, DMSO is used as a negative control, and resveratrol (RES) is used as a positive control. Incubate at 37°C, 200rpm for 24h, and centrifuge at 12,000rpm, 4°C for 10min.

[0029] Determination of protease activity: Prepare 2% azo casein solution (weigh 100mg azo casein and dissolve in 5mL Tris / HCl with a concentration of 50mM, store at 4°C for later use), take 250uL of this solution and mix with 150uL PA01 filtrate, Place at 4°C for 4h, then add 1.2mL of 10% trichloroacetic acid to stop the reaction, mix well and let stand at 4°C for 15min, then centrifuge at 10,000rpm for 10min, discard the precipitate,...

Embodiment 3

[0044] Influence on the expression of quorum sensing related genes: Inoculate a single colony of Pseudomonas aeruginosa PAO1 into NB medium, culture overnight at 37°C and 150rmp. Tune OD 620 = 0.5, take 50 μL of the OD-adjusted bacterial solution and add it to 50 mL NB medium at a ratio of 1:1000, and add different concentrations of hordenine at the same time, and use DMSO as a negative control. Cultivate at 37°C and 200rpm for 24h. Total RNA was extracted according to the method introduced by the RNeasy Mini Kit kit (Tiangen Bio, Beijing), RNA reverse transcription experiments were performed according to the method introduced by the RNeasy Mini Kit kit, and primers for lasI, lasR, rhlI, rhlR and the internal reference gene rpsL were set (As shown in Table 1)

[0045] Table 1 Experimental primers

[0046] Table 1Primers used in this study

[0047]

[0048] (1) According to Premix Ex TaqTM (Tli RnaseH Plus) kit to establish qRT-PCR reaction system, as shown in Table 2 ...

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Abstract

The invention discloses new application of a natural product hordenine and in particular relates to application of a compound hordenine in preparation of a medicine used for preventing and treating bacterial infection. The new application is mainly reflected in that hordenine can obviously inhibit synthesis of a quorum sensing-regulated signal molecule, hordenine can obviously inhibit expression of a quorum sensing-regulated virulence factor and hordenine can obviously inhibit expression of a quorum sensing-related gene. Besides, drug combination of hordenine and antibiotic netilmicin can obviously inhibit generation of a biofilm, a formed biofilm can be obviously destroyed, and the hordenine is hopeful to becoming a novel anti-infective agent for preventing and treating bacterial infection.

Description

technical field [0001] The invention belongs to medical technology, and in particular relates to the anti-infection pharmaceutical use of hordenine or its combined antibiotics. Background technique [0002] Hordenine (CAS Registry No.: 539-15-1) is a natural phenolic compound that exists in the roots of germinated barley seeds, the roots of leguminous plants, the stems of Limeleaf leeches, the The heartwood of pigeonpea can also be artificially synthesized and has been commercialized. Its structural formula is as follows: [0003] [0004] Hordenine is relatively safe and has the effects of relaxing bronchial smooth muscle, constricting blood vessels, pressurizing blood vessels, raising blood pressure and stimulating the central nervous system, and can be used to relieve bronchitis and bronchial asthma. But so far, there is no relevant report about the anti-infection research of hordenine. [0005] Due to the abuse of antibiotics, more and more multi-drug resistant str...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/06A61K31/137A61K31/7048A61P31/04
CPCA61K31/137A61K31/7048A61K45/06A61K2300/00
Inventor 贾爱群周金伟程炜佳
Owner HAINAN UNIV
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