Composition for inhibiting formation of staphylococcus aureus biofilm and application thereof

Abstract

The invention provides a composition for inhibiting formation of a staphylococcus aureus biofilm, and relates to the technical field of biological medicines, and the composition comprises an inhibitor and vancomycin with sub-inhibitory concentration. In order to solve the clinical problem that vancomycin with sub-inhibitory concentration promotes formation of a staphylococcus aureus biofilm, the invention provides an inhibition method with very high feasibility. Through joint use of the vancomycin with the sub-inhibitory concentration and the arginine, formation of a staphylococcus aureus biofilm can be effectively inhibited, the treatment effect of the vancomycin with the sub-inhibitory concentration is improved, the treatment dosage of the vancomycin is expected to be reduced, and drug resistance generated by bacteria is avoided.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a composition for inhibiting the formation of Staphylococcus aureus biofilm and application thereof. Background Art

[0002] Bacteria can be exposed to sub-inhibitory concentrations of antibiotics at the beginning and end of dosing or during normal-dose versus low-dose therapy. Sub-inhibitory concentrations, i.e., antibiotic concentrations below the minimum inhibitory concentration (MIC), can have a variety of effects on bacteria, such as making them more capable of biofilm formation. There is growing evidence that sub-inhibitory concentrations of antimicrobial agents can stimulate bacterial biofilm formation and alter the composition of the biofilm matrix, leading to an increase in biofilm infections.

[0003] Biofilm infection (BFI) refers to an infection caused by pathogens forming biofilms on the surface of human tissues or medical devices. In recent years, with the increase in the use of indwelling medical devices and biomaterials, the incidence of biofilm-related infections has increased year by year. Staphylococcus aureus (abbreviated as S. aureus) accounts for 34% of all causes of biomaterial-related infections and is known as a powerful biofilm producer. Vancomycin is the last line of drug for the treatment of S. aureus infections, but due to the drug's metabolic dynamics and the complex environment in the body, it is often difficult for vancomycin to reach an effective inhibitory concentration, exposing S. aureus to sub-inhibitory concentrations. For example, during surgical operations, S. aureus is at greater risk of being exposed to sub-inhibitory concentrations of vancomycin because the bioavailability of vancomycin in tissues is lower than that of other antibiotics.

[0004] Although the phenomenon that sub-inhibitory concentration vancomycin promotes the formation of biofilm of Staphylococcus aureus has been confirmed by many studies, there are few studies on the mechanism of this phenomenon. At present, the main studies include changes in extracellular vesicles and mutations in agr genes. Other similar mechanisms of sub-inhibitory concentration antibiotics promoting bacterial biofilm formation include increased eDNA secretion, obstruction of antibiotic penetration, and changes in biofilm formation-related genes such as cidA. However, due to the limitations of experimental techniques and methods, the research on these mechanisms is not in-depth, and therefore no practical solution is provided to solve the problem of sub-inhibitory concentration vancomycin promoting the formation of biofilm of Staphylococcus aureus. Summary of the invention

[0005] 1. Technical issues to be resolved

[0006] In view of the shortcomings of the prior art, the present invention provides a composition for inhibiting the formation of biofilm of Staphylococcus aureus and its application, which solves the problem that sub-inhibitory concentration of vancomycin promotes the formation of biofilm of Staphylococcus aureus, leading to enhanced drug resistance of Staphylococcus aureus and then reducing the treatment effect of bacterial infection.

[0007] (II) Technical solution

[0008] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0009] In one aspect, the present invention provides the use of arginine in the preparation of a medicament for inhibiting the promotion of biofilm formation of Staphylococcus aureus by vancomycin at a sub-inhibitory concentration.

[0010] In one aspect, the present invention provides a composition for inhibiting the biofilm formation of Staphylococcus aureus, comprising an effective amount of arginine and a sub-inhibitory concentration of vancomycin.

[0011] In one embodiment, the added amount of arginine is 50-150 mM.

[0012] In one embodiment, the MIC value is 1 ug / ml.

[0013] In one embodiment, the sub-inhibitory concentration of vancomycin is added in an amount of 1 / 2 MIC.

[0014] In one aspect, the present invention provides a pharmaceutical composition for inhibiting the growth of Staphylococcus aureus, wherein the composition comprises a therapeutically effective amount of arginine and a sub-inhibitory concentration of vancomycin.

[0015] In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0016] In one embodiment, the added amount of arginine is 50-150 mM.

[0017] In one embodiment, the amount of sub-inhibitory concentration of vancomycin added is greater than 1 / 10 MIC and less than 1 MIC.

[0018] In one embodiment, the sub-inhibitory concentration of vancomycin is added in an amount of 1 / 2 MIC.

[0019] In one embodiment, the MIC value is 1 ug / ml.

[0020] In one aspect, the present invention provides the use of the above-mentioned composition in the preparation of a drug for inhibiting the growth of Staphylococcus aureus.

[0021] In one aspect, the present invention provides a method for inhibiting the formation of Staphylococcus aureus biofilm, using any of the inhibitors described above, and when using sub-inhibitory concentrations of vancomycin to treat Staphylococcus aureus infection, simultaneously using the inhibitor to weaken the ability of sub-inhibitory concentrations of vancomycin to promote the formation of Staphylococcus aureus biofilm, wherein the inhibitor includes a therapeutically effective amount of arginine.

[0022] In one embodiment, the Staphylococcus aureus is Staphylococcus aureus strain 8325.

[0023] (III) Beneficial effects

[0024] The present invention provides a composition for inhibiting the formation of Staphylococcus aureus biofilm and its application. Compared with the prior art, it has the following beneficial effects:

[0025] 1. The present invention uses in vitro experiments to show that after exogenous addition of arginine or overexpression of arginine synthesis genes, the biofilm formation of Staphylococcus aureus under the pressure of sub-inhibitory concentration vancomycin is significantly reduced. qrt-PCR experiments and bacterial adhesion experiments show that arginine inhibits bacterial adhesion and thus weakens the ability of Staphylococcus aureus to form biofilm under the pressure of sub-inhibitory concentration vancomycin.

[0026] 2. The present invention provides a highly feasible inhibition method to solve the clinical problem that sub-inhibitory concentration vancomycin promotes the formation of Staphylococcus aureus biofilm. By constructing an in vivo experiment of a mouse catheter infection model, it is proved that the use of sub-inhibitory concentration vancomycin and arginine together can effectively inhibit the formation of Staphylococcus aureus biofilm, reduce the amount of bacteria attached to the catheter, and enhance the therapeutic effect of sub-inhibitory concentration vancomycin, which is expected to provide a reference for reducing the therapeutic dosage of vancomycin and the development of bacterial resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Figure 1 This is a data diagram of the microporous film forming method and laser confocal laser experiment in Example 1 of the present invention;

[0029] Among them, (a) is a crystal violet staining image after static culture of Staphylococcus aureus 832 in a 96-well plate using BHI medium for 56 hours, 12 hours, and 20 hours;

[0030] (b) is a bar graph of the OD560 values ​​of each well after crystal violet staining of the biofilm of Staphylococcus aureus strain 8325;

[0031] (c) The biofilm structure of Staphylococcus aureus was observed using a laser confocal microscope after treatment with 1 / 2MIC vancomycin (i.e., sub-inhibitory concentration of vancomycin) for 12 h and 20 h;

[0032] (d) is a diagram showing the biofilm formation of clinical strains of Staphylococcus aureus after treatment with 1 / 2MIC vancomycin (i.e., sub-inhibitory concentration of vancomycin) using the crystal violet staining method (microporous film formation method);

[0033] (e) is the OD560 value graph corresponding to each well of different strain samples in (d).

[0034] Figure 2 The data graphs are of the biofilm enzymatic hydrolysis experiment, biofilm matrix component determination experiment, adhesion bacteria experiment and autolysis experiment in Example 2 of the present invention;

[0035] Among them, (a) is a graph showing the residual amount of biofilm after proteinase K, deoxyribonuclease I (DNase I) and sodium periodate were used to decompose the protein, eDNA and polysaccharide components in the biofilm of Staphylococcus aureus respectively by crystal violet staining method;

[0036] (b) The OD560 value of each well after crystal violet staining of the biofilm of Staphylococcus aureus strain 8325;

[0037] (c)-(e) are experimental diagrams for determining the components of biofilm matrix, (c) is the content of extracellular protein in the biofilm of Staphylococcus aureus strain 8325; (d) is the content of eDNA in the biofilm of Staphylococcus aureus strain 8325; (e) is the content of PIA (polysaccharide) in the biofilm of Staphylococcus aureus strain 8325;

[0038] (f)-(g) are the initial adherent bacterial counts and statistical graphs of Staphylococcus aureus strain 8325 after 0.5 h and 1 h of vancomycin treatment;

[0039] (h) is the eDNA autolysis assay of Staphylococcus aureus strain 8325 after vancomycin treatment for 12 h, and the autolysis activity is monitored by the change of OD630 over time;

[0040] (i) Real-time fluorescence quantitative PCR was used to detect the changes in transcription levels of biofilm-related genes in Staphylococcus aureus strain 8325 under the action of vancomycin.

[0041] Figure 3 This is a metabolomics analysis data diagram of Example 3 of the present invention;

[0042] Among them, (a) is the PCA score graph;

[0043] (b) is a classification diagram of metabolites in the HMDB database, which is displayed from high to low according to the percentage of metabolites. Different types are represented by different colors, and the size of the fan represents the percentage.

[0044] (c) Volcano plot of differential metabolites between the control group (BHI) and the treatment group (BHI-VAN);

[0045] (d) is the Kegg pathway enrichment diagram of differential metabolites, the vertical axis represents the enrichment rate, the horizontal axis represents the pathway name, and the darker the column color, the more significant it is;

[0046] (e) shows the top 30 differential metabolites with the highest VIP values. The bar chart of the VIP values ​​of the metabolites is on the far right. The color of the bar indicates the P value. The darker the color, the smaller the P value.

[0047] Figure 4 This is a transcriptomics analysis data diagram of Example 3 of the present invention;

[0048] Among them, (a) is Figure 3 (e) PCA score plot of transcriptome data of six samples;

[0049] (b) Scatter plot of DEGs (differentially expressed genes) between the control group (BHI) and the treatment group (BHI-VAN);

[0050] (c) To visualize and cluster the enriched DEG terms in the GO database using the Cytoscape plugin ClueGO, based on P < 0.05 and κ score ≥ 0.1;

[0051] (d) is a pie chart showing the proportions of different clusters;

[0052] (e) Protein-protein interaction network (PPI) diagram of DEGs;

[0053] (f) KEGG enrichment analysis of DEGs;

[0054] (g) RT-qPCR analysis of five selected DEGs; RT-qPCR results are the average of -ΔΔCT obtained from three biological replicates, and the error bars represent the standard deviation. The results were normalized using gyrB and expressed as fold change.

[0055] Figure 5 This is a graph showing the combined analysis of metabolome and transcriptome in Example 3;

[0056] Among them, (a) is a quadrant diagram of the association between DAMs and DEGs in the control group (BHI) and the treatment group (BHI-VAN); the horizontal axis represents the log2 ratio of genes, and the vertical axis represents the log2 ratio of metabolites; the black dotted line represents different thresholds; each point represents a gene or metabolite; black dots represent unchanged genes or metabolites; green dots represent unchanged genes but differentially accumulated metabolites; red dots represent differentially expressed genes of unchanged metabolites; blue dots represent both differentially expressed genes and differentially accumulated metabolites; from left to right and from top to bottom, it is divided into quadrants 1 to 9 by black dotted lines; quadrants 1, 2, and 4 indicate that the expression abundance of metabolites is higher than that of genes; quadrants 3 and 7 indicate that the expression pattern of genes is consistent with that of metabolites; quadrant 5 indicates that there is no differential expression of genes and metabolites; quadrants 6, 8, and 9 indicate that the expression abundance of metabolites is lower than that of genes;

[0057] (b) KEGG pathway annotation diagram of DEGs in quadrants 7 and 9;

[0058] (c) Arginine biosynthesis pathway via glutamate and proline, respectively.

[0059] Figure 6 The graph is a graph showing the inhibition of biofilm formation of Staphylococcus aureus under different additive culture conditions of Examples 4-5 of the present invention and Comparative Examples 1-6;

[0060] Among them, (a) is a crystal violet staining image of the biofilm formation of Staphylococcus aureus under different additive culture conditions;

[0061] (b) is a graph showing the biofilm formation rate (OD560) of Staphylococcus aureus strains under different concentrations of arginine added;

[0062] (c) is a graph showing the relative expression level of the argG gene in the S. aureus strain 8325-pli50-argG;

[0063] (d) Crystal violet staining of the biofilm formation of the S. aureus strain overexpressing argG (8325-pli50-argG) under the action of 1 / 2MIC vancomycin;

[0064] (e) is a graph showing the effect of 1 / 2 MIC vancomycin on 8325-pli50-argG (OD560) biofilm formation;

[0065] (f) is the relative expression level of biofilm-related genes in the control group PBS, 1 / 2MIC vancomycin treatment group, 50mM arginine treatment group, and vancomycin-arginine treatment group after treatment of Staphylococcus aureus for 12 hours;

[0066] (g) is a bar graph showing the effects of vancomycin and arginine on bacterial adhesion determined by plate count method;

[0067] (h) Bar graph showing the effects of vancomycin and argG gene overexpression on bacterial adhesion ability as determined by the plate count method.

[0068] Figure 7 This is a data diagram of an in vivo experiment conducted by constructing a mouse catheter infection model in Example 6 of the present invention;

[0069] Among them, (a) is the experimental flow chart of mouse catheter infection model;

[0070] (b) is a SEM observation of the microstructure of the biofilm on the catheter;

[0071] (c) is a graph showing the results of the experimental and control groups in treating mouse catheter Staphylococcus aureus infection. DETAILED DESCRIPTION

[0072] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0073] The embodiments of the present invention provide a composition for inhibiting the formation of biofilm of Staphylococcus aureus and its application, thereby solving the problem that sub-inhibitory concentration vancomycin promotes the formation of biofilm of Staphylococcus aureus, leading to enhanced drug resistance of Staphylococcus aureus, and then leading to decreased treatment effect of bacterial infection. The present invention proves that the use of sub-inhibitory concentration vancomycin and arginine together can effectively inhibit the formation of biofilm of Staphylococcus aureus, reduce the amount of bacteria attached to the catheter, and enhance the therapeutic effect of sub-inhibitory concentration vancomycin, which is expected to provide reference significance for reducing the therapeutic dosage of vancomycin and the generation of bacterial resistance.

[0074] A composition for inhibiting the biofilm formation of Staphylococcus aureus comprises an inhibitor and sub-inhibitory concentration of vancomycin.

[0075] In some embodiments, the inhibitor is arginine or overexpression of arginine synthesis genes.

[0076] In some specific embodiments, the amount of arginine added is 50-150 mM.

[0077] In some embodiments, the overexpressed arginine synthesis gene is argG or argH.

[0078] In some specific embodiments, the amount of sub-inhibitory concentration of vancomycin added is 1 / 2 MIC.

[0079] The method for inhibiting the formation of Staphylococcus aureus biofilm comprises the following steps: when using sub-inhibitory concentration vancomycin to treat Staphylococcus aureus infection, simultaneously using an inhibitor to reduce the ability of sub-inhibitory concentration vancomycin to promote the formation of Staphylococcus aureus biofilm.

[0080] The MIC referred to in the present invention is the minimum inhibitory concentration of vancomycin. The MIC value may be different for different Staphylococcus aureus, and the MIC value can be determined by industry technicians through conventional means.

[0081] In some embodiments, the S. aureus strain is S. aureus strain 8325.

[0082] In order to better understand the above technical solution, the above technical solution will be described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0083] Embodiment 1:

[0084] The formation of biofilm of Staphylococcus aureus under the action of sub-inhibitory concentration of vancomycin was observed by microporous film formation method and laser confocal experiment, which proved that sub-inhibitory concentration of vancomycin can promote the formation of biofilm of standard strain of Staphylococcus aureus. The experimental data are as follows Figure 1 shown.

[0085] Depend on Figure 1 (a)-(b) It can be seen that the longer the culture time of S. aureus strain 8325 is, and when the amount of vancomycin added is 0.4-0.95 μg / ml, the more S. aureus biofilm is formed.

[0086] Depend on Figure 1 (c)-(e) It can be seen that 1 / 2MIC vancomycin (i.e., sub-inhibitory concentration of vancomycin) promotes the formation of S. aureus biofilm.

[0087] Embodiment 2:

[0088] Through the biofilm enzymatic hydrolysis experiment and the biofilm matrix component determination experiment, combined with Figure 2 (a)-(e) It can be found that the protein and eDNA components in the S. aureus biofilm matrix increased significantly under the pressure of sub-inhibitory concentration of vancomycin.

[0089] Through relevant phenotypic experiments (adhesion bacteria experiment and autolysis experiment), combined with Figure 2 (f)-(i) further demonstrated that subinhibitory concentrations of vancomycin enhanced the formation of S. aureus biofilm by promoting bacterial adhesion and autolysis.

[0090] Embodiment 3:

[0091] By using metabolomics data, transcriptomics data and joint analysis, the potential regulatory mechanism of enhanced S. aureus biofilm formation under the action of subinhibitory concentration of vancomycin was revealed, and the key regulatory factors were screened out.

[0092] Depend on Figure 3 It can be seen that the metabolomics data results show that under the action of sub-inhibitory concentration of vancomycin, Staphylococcus aureus has a total of 115 differential metabolites (dam), and these differential metabolites (dam) are enriched in various amino acid metabolic processes, indicating that sub-inhibitory concentration of vancomycin affects the amino acid metabolism of Staphylococcus aureus.

[0093] Depend on Figure 4 It can be seen that the transcriptomics data results show that under the action of subinhibitory concentration of vancomycin, Staphylococcus aureus has 319 differentially expressed genes (DEGs), and 15 biofilm formation-related genes were screened out among these DEGs, mainly involving the expression of adhesion proteins, bacterial autolysis and biofilm dissociation, which is consistent with the experimental results of Examples 1 and 2.

[0094] KEGG gene pathway enrichment analysis showed that DEGs were enriched in the arginine biosynthesis pathway. By constructing a protein interaction network encoded by differentially expressed genes, we screened out core genes that changed under the action of subinhibitory concentrations of vancomycin, including arginine synthesis genes (argG, argH) and arginine decomposition genes (arcR, arcA, arcC, nos).

[0095] Depend on Figure 5 Correlation analysis was performed between dam and DEGs, and a nine-quadrant diagram was drawn to screen out DEGs significantly correlated with dam, which were annotated to the arginine synthesis pathway, suggesting that subinhibitory concentration of vancomycin promotes the formation of S. aureus biofilm by inhibiting arginine synthesis.

[0096] Embodiment 4:

[0097] The method for inhibiting the biofilm formation of Staphylococcus aureus comprises the following steps: using a sub-inhibitory concentration of vancomycin-arginine combination, and culturing the cells in a 96-well plate in a TSB medium containing 2% glucose for 20 hours.

[0098] Embodiment 5:

[0099] The method for inhibiting the formation of Staphylococcus aureus biofilm comprises adding an overexpressed argG gene to the Staphylococcus aureus strain, adding a sub-inhibitory concentration of vancomycin to the culture medium, and culturing the culture medium in a 96-well plate in a TSB medium containing 2% glucose for 20 hours.

[0100] Comparative Example 1 is different from Example 4 in that:

[0101] Under the condition of using PBS, the cells were cultured in TSB medium containing 2% glucose in a 96-well plate for 20 h.

[0102] Comparative Example 2 is different from Example 4 in that:

[0103] Under the condition of using sub-inhibitory concentration of vancomycin (1 / 2MIC), the cells were cultured in TSB medium containing 2% glucose in a 96-well plate for 20 h.

[0104] Comparative Example 3 is different from Example 4 in that:

[0105] The cells were cultured in a 96-well plate in TSB medium supplemented with 2% glucose for 20 h in the presence of 5 mM arginine.

[0106] Comparative Example 4 is different from Example 4 in that:

[0107] The cells were cultured in a 96-well plate in TSB medium supplemented with 2% glucose for 20 h in the presence of 50 mM arginine.

[0108] Comparative Example 5 is different from Example 4 in that:

[0109] The cells were cultured in a 96-well plate in TSB medium supplemented with 2% glucose for 20 h in the presence of 150 mM arginine.

[0110] Comparative Example 6 is different from Example 5 in that:

[0111] The method for inhibiting the formation of Staphylococcus aureus biofilm comprises adding an overexpressed argG gene to the Staphylococcus aureus strain and culturing the strain in a 96-well plate in a TSB medium containing 2% glucose for 20 hours.

[0112] Figure 6 The graphs are for the inhibition of biofilm formation of Staphylococcus aureus under different additive culture conditions of Examples 4-5 and Comparative Examples 1-6. Figure 6 It can be seen that under the conditions of using a sub-inhibitory concentration of vancomycin-arginine combination or adding an overexpressed argG gene to the S. aureus strain and adding sub-inhibitory concentration of vancomycin to the culture medium, the biofilm formation of S. aureus was significantly inhibited and the therapeutic effect of vancomycin was improved.

[0113] This indicates that when using sub-inhibitory concentrations of vancomycin, adding exogenous arginine or overexpressing the arginine synthesis gene (argG) can offset the ability of sub-inhibitory concentrations of vancomycin to promote the formation of biofilms of Staphylococcus aureus. The results of qRT-PCR experiments showed that the transcriptional expression of bacterial adhesion-related genes decreased significantly after exogenous addition of arginine, and phenotypic experiments also confirmed that arginine weakened the adhesion ability of Staphylococcus aureus, indicating that arginine weakened the ability of Staphylococcus aureus to form biofilms under the action of sub-inhibitory concentrations of vancomycin by inhibiting bacterial adhesion.

[0114] Embodiment 6:

[0115] The in vivo effect of the arginine-subinhibitory concentration vancomycin combination was analyzed by constructing a mouse catheter infection model.

[0116] The specific operation method of the mouse catheter infection model experiment is as follows:

[0117] Female BALB / c mice aged 9-11 weeks were used and mice received drug treatment every 24 h starting from day 1 after infection and continuing for 3 days until the end of the experiment.

[0118] The control group was injected with normal saline.

[0119] The experimental groups included:

[0120] (1) Inject 0.2 ml of vancomycin containing 0.0625 mg / ml (0.625 mg / kg) via the tail vein;

[0121] (2) Inject 0.2 ml of arginine containing 0.02 mg / ml (or 0.2 mg / kg) through the tail vein;

[0122] (3) Inject 0.1 ml of a combination of 0.125 mg / ml vancomycin and 0.1 ml of 0.04 mg / ml arginine through the tail vein.

[0123] SEM was used to observe the microstructure of the biofilm on the catheter. Figure 7 The results showed that subinhibitory concentrations of vancomycin could promote the formation of biofilms by Staphylococcus aureus on catheters in vivo, but simultaneous treatment with arginine significantly reduced the formation of biofilms by Staphylococcus aureus on catheters.

[0124] In summary, compared with the prior art, the present invention has the following beneficial effects:

[0125] 1. Overexpression of arginine synthesis gene can be used to synthesize arginine, which is an important amino acid. It is commonly used clinically to treat some diseases and can also be used as a nutritional supplement to promote metabolism and growth and development without serious adverse reactions. The present invention has shown through in vitro experiments that after exogenous addition of arginine or overexpression of arginine synthesis gene, the formation of Staphylococcus aureus biofilm under the pressure of sub-inhibitory concentration vancomycin is significantly reduced. qrt-PCR experiments and bacterial adhesion experiments show that arginine inhibits bacterial adhesion and thus weakens the ability of Staphylococcus aureus to form biofilm under the pressure of sub-inhibitory concentration vancomycin.

[0126] 2. The present invention provides a highly feasible inhibition method to solve the clinical problem that sub-inhibitory concentration vancomycin promotes the formation of Staphylococcus aureus biofilm. By constructing an in vivo experiment of a mouse catheter infection model, it is proved that the use of sub-inhibitory concentration vancomycin and arginine together can effectively inhibit the formation of Staphylococcus aureus biofilm, reduce the amount of bacteria attached to the catheter, and enhance the therapeutic effect of sub-inhibitory concentration vancomycin, which is expected to provide a reference for reducing the therapeutic dosage of vancomycin and the development of bacterial resistance.

[0127] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0128] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of arginine in the preparation of drugs that inhibit the formation of biofilm of Staphylococcus aureus promoted by vancomycin at sub-inhibitory concentrations.

2. A composition for inhibiting the formation of Staphylococcus aureus biofilm, It is characterized in that The composition includes an effective amount of arginine and a sub-inhibitory concentration of vancomycin.

3. The composition according to claim 2, It is characterized in that The added amount of arginine is 50-150 mM.

4. The composition according to claim 2, It is characterized in that The added amount of the sub-inhibitory concentration of vancomycin is greater than 1 / 10 MIC and less than 1 MIC.

5. A pharmaceutical composition for inhibiting the growth of Staphylococcus aureus, It is characterized in that The composition includes a therapeutically effective amount of arginine and a sub-inhibitory concentration of vancomycin.

6. The pharmaceutical composition according to claim 5, It is characterized in that The added amount of arginine is 50-150 mM.

7. The pharmaceutical composition according to claim 5, It is characterized in that The added amount of the sub-inhibitory concentration of vancomycin is greater than 1 / 10 MIC and less than 1 MIC.

8. The pharmaceutical composition according to claim 7, It is characterized in that The added amount of the sub-inhibitory concentration of vancomycin is 1 / 2 MIC.

9. The pharmaceutical composition according to claim 5, It is characterized in that The pharmaceutical composition also includes pharmaceutically acceptable excipients.

10. Use of the composition for inhibiting the biofilm formation of Staphylococcus aureus according to any one of claims 2 to 4 or the pharmaceutical composition according to any one of claims 5 to 9 in the preparation of a drug for inhibiting the growth of Staphylococcus aureus.

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