Use of anisaldehyde / meropenem or their combination in the preparation of a medicament for preventing and / or treating Erwinia infection

Abstract

The present invention discloses the use of anisaldehyde / meropenem or their combination in the preparation of a medicament for preventing and / or treating Erwinia infection; the present invention discovers for the first time that anisaldehyde has an inhibitory effect on the formation of Erwinia biofilm; meropenem has an inhibitory effect on the growth of Erwinia; the combination of anisaldehyde and meropenem can effectively inhibit the formation of Erwinia biofilm; and anisaldehyde can enhance the drug sensitivity of meropenem to Erwinia, indicating a good application prospect in agricultural medicaments for preventing and / or treating Erwinia infection.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical technology, and in particular to the use of anisic aldehyde / meropenem or their combination in the preparation of a drug for preventing and / or treating Erwinia infection. Background Art

[0002] Bacterial soft rot is a plant disease widely existing around the world, with a wide range of host plants, mainly including food crops and vegetables. Bacterial soft rot is mainly caused by Erwinia, and the spoilage of food crops and vegetables caused by Erwinia has caused serious economic losses. Using chemical fungicides is the main way to control Erwinia infection. However, the long-term use of chemical fungicides has caused the bacteria to develop drug resistance on the one hand, and on the other hand, their drug residues have also posed potential hazards to human health. How to reduce the drug resistance of Erwinia and reduce the use amount of chemical fungicides is a major problem faced by the current scientific community.

[0003] The formation of biofilms is one of the important reasons for the drug resistance of Erwinia. Inhibiting the formation of Erwinia biofilms is an important way to reduce the drug resistance of this bacterium. Seeking the combined use of biofilm inhibitors and traditional chemical fungicides can not only reduce the drug resistance of Erwinia, but also reduce the dosage of chemical fungicides, which is an important solution for preventing and treating Erwinia infection.

[0004] Anisic aldehyde (CAS registration number: 123-11-5) is an aldehyde substance extracted from the spice star anise, and its structure is as follows:

[0005]

[0006] Anisic aldehyde has antibacterial and anti-tumor activities, but so far there has been no research on the prevention and treatment of Erwinia infection with anisic aldehyde.

[0007] Meropenem (CAS registration number: 96036-03-2) is a carbapenem antibiotic with good antibacterial activity against common clinically Gram-negative and Gram-positive bacteria, and is commonly used to treat infections induced by bacteria sensitive to this drug. Its structure is as follows:

[0008] Summary of the Invention

[0009] Object of the Invention: The object of the present invention is to provide the use of anisic aldehyde / meropenem or their combination in the preparation of a drug for preventing and / or treating Erwinia infection.

[0010] Technical Solution: The present invention relates to the use of anisic aldehyde / meropenem in the preparation of a drug for preventing and / or treating Erwinia infection.

[0011] Furthermore, anisaldehyde achieves the purpose of preventing and / or treating infections by inhibiting the formation of Erwinia biofilms, and the final concentration of anisaldehyde is 0.5 - 1 μL / mL.

[0012] Furthermore, meropenem inhibits the growth of Erwinia to achieve the purpose of preventing and / or treating infections, and the MIC of meropenem is 0.5 μg / mL.

[0013] The present invention also provides an application of anisaldehyde combined with meropenem in the preparation of a drug for preventing and / or treating Erwinia infections.

[0014] Furthermore, anisaldehyde combined with meropenem achieves the purpose of preventing and / or treating infections by inhibiting the growth of Erwinia.

[0015] Furthermore, in the case where anisaldehyde combined with meropenem achieves the purpose of preventing and / or treating infections by inhibiting the growth of Erwinia, the final concentration of anisaldehyde is 0.5 - 1 μL / mL, and the final concentration of meropenem is 0.125 - 0.5 μg / mL.

[0016] Furthermore, in the case where anisaldehyde combined with meropenem achieves the purpose of preventing and / or treating infections by inhibiting the growth of Erwinia, the final concentration of anisaldehyde is 0.25 - 1 μL / mL, and the final concentration of meropenem is 0.25 - 0.5 μg / mL.

[0017] Furthermore, anisaldehyde combined with meropenem achieves the purpose of preventing and / or treating infections by inhibiting the formation of Erwinia biofilms.

[0018] Furthermore, in the case where anisaldehyde combined with meropenem achieves the purpose of preventing and / or treating infections by inhibiting the formation of Erwinia biofilms, the final concentration of anisaldehyde is 0.5 - 1 μL / mL, and the final concentration of meropenem is 0.125 - 0.25 μg / mL.

[0019] Furthermore, in the case where anisaldehyde combined with meropenem achieves the purpose of preventing and / or treating infections by inhibiting the formation of Erwinia biofilms, the final concentration of anisaldehyde is 0.5 - 1 μL / mL, and the final concentration of meropenem is 0.25 μg / mL.

[0020] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages:

[0021] (1) Anisaldehyde has an inhibitory effect on the formation of Erwinia biofilms; meropenem has an inhibitory effect on the growth of Erwinia.

[0022] (1) Anisaldehyde can enhance the drug sensitivity of meropenem to Erwinia.

[0023] (2) Anisaldehyde combined with meropenem can effectively inhibit the formation of Erwinia biofilm, indicating good prospects for agricultural drug applications in preventing and / or treating Erwinia infections. Description of the Drawings

[0024] Figure 1 Inhibitory effect of anisaldehyde / meropenem or their combination on the growth of Erwinia;

[0025] Figure 2 Inhibitory effect of anisaldehyde / meropenem or their combination on the formation of Erwinia biofilm. Detailed Embodiments

[0026] The technical solutions of the invention will be further described below in conjunction with the embodiments.

[0027] Example 1

[0028] Inhibitory effect on the growth of Erwinia:

[0029] Drugs: Anisaldehyde was isolated and extracted from the spice plant Illicium verum. Anisaldehyde used by those skilled in the art can be directly purchased from Shanghai Yuanye Bio-Technology Co., Ltd. Meropenem can be directly purchased from Shanghai Yuanye Bio-Technology Co., Ltd.

[0030] Strain: Erwinia was gifted by Professor Yao Weirong of Jiangnan University.

[0031] Pick a single colony of Erwinia and inoculate it into LB medium, and culture it at 28 °C and 180 rpm for 17 h. Take 5 μL of the cultured bacterial solution and inoculate it into 5 mL of fresh LB medium. At the same time, add anisaldehyde (Ana) with final concentrations of 0.25, 0.5, 1 μL / mL, meropenem with final concentrations of 0.03125, 0.0625, 0.125, 0.25, 0.5 μg / mL, and combined medications of anisaldehyde and meropenem at different concentrations. Use DMSO instead of meropenem as the negative control. Among them, -Ana represents an anisaldehyde concentration of 0 μL / mL, and culture at 28 °C and 180 rpm for 12 h. The cultured bacterial solution was measured by a multifunctional microplate reader for OD 600 to determine the effects of the drugs and drug combinations on the growth of Erwinia.

[0032] Test results: As Figure 1 shown, the results of the DMSO control group showed that the inhibitory effect of anisaldehyde alone at concentrations of 0.25, 0.5, and 1 μL / mL on the growth of Erwinia was weak. The minimum inhibitory concentration (MIC) of meropenem alone against Erwinia was 0.5 μg / mL.

[0033] When meropenem was combined with anisaldehyde at a concentration of 1 μL / mL, the MIC of meropenem decreased from 0.5 μg / mL to 0.125 μg / mL; although anisaldehyde at a concentration of 0.5 μL / mL could not reduce the MIC value of meropenem against Erwinia, it could significantly enhance the inhibitory effect of meropenem on Erwinia.

[0034] Example 2

[0035] Inhibitory effect on the formation of Erwinia biofilm:

[0036] Pick a single colony of Erwinia and inoculate it into LB medium, and culture it at 28 °C and 180 rpm for 17 h. Take the cultured bacterial solution and inoculate it into TSB medium (containing 1% glycerol) at an inoculation amount of 0.1%. After mixing, add anisaldehyde with final concentrations of 0.5 and 1 μL / mL, meropenem with final concentrations of 0.125 and 0.25 μg / mL, and combined use of anisaldehyde and meropenem at different concentrations. Use DMSO instead of meropenem as the negative control, where -Ana indicates that the concentration of anisaldehyde is 0 μL / mL. Take 1 mL of the mixed bacterial solution from each treatment group and add it to a 24-well plate, and culture it statically at 28 °C for 24 h. Aspirate the supernatant, wash it 3 times with PBS to wash away the floating bacteria, place it in an oven at 60 °C to dry the moisture, add 200 μL of methanol to fix it for 15 min. Aspirate the methanol and dry it in an oven at 60 °C. Add 100 μL of 0.05% crystal violet and stain it for 15 min. Aspirate the crystal violet, wash it 3 times with PBS to remove the excess unattached crystal violet dye, and place it in an oven at 60 °C to dry. After drying, add 200 μL of 95% ethanol, and decolorize it in a shaker at 37 °C and 180 rpm for 15 min. Take 150 μL of the decolorized solution and measure the OD with an enzyme-linked immunosorbent assay instrument. 570 .

[0037] Test results: As Figure 2 shown, the results of the DMSO control group showed that when treated with anisaldehyde alone at concentrations of 0.5 and 1 μL / mL, the formation of Erwinia biofilm could be significantly inhibited, and the inhibition rates were 20% and 31% respectively.

[0038] When meropenem was used alone at concentrations of 0.125 and 0.25 μg / mL, it had no inhibitory effect on the biofilm.

[0039] When meropenem was combined with anisaldehyde, its inhibitory effect on Erwinia biofilm was significantly enhanced. When anisaldehyde was combined with meropenem at a concentration of 0.125 μg / mL, the inhibition rates of the biofilm were 51% and 86% respectively, and when anisaldehyde was combined with meropenem at a concentration of 0.25 μg / mL, the inhibition rates of the biofilm increased to 88% and 93%.

Claims

1. Use of anisaldehyde combined with meropenem in the preparation of a medicament for preventing and / or treating Erwinia infection, characterized in that, The final concentration of anisaldehyde is 0.5 - 1 μL / mL, and the final concentration of meropenem is 0.125 - 0.25 μg / mL.

2. The use according to claim 1, characterized in that, The combination of anisaldehyde and meropenem inhibits the growth of Erwinia bacteria.

3. The use according to claim 1, characterized in that, The combination of anisaldehyde and meropenem inhibits the formation of Erwinia biofilms.

4. The use according to claim 1, characterized in that, The final concentration of anisaldehyde is 0.5 - 1 μL / mL, and the final concentration of meropenem is 0.25 μg / mL.

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