Study On The Antibacterial Mechanism Of Genistein And The Preparation And Characterization Of Microemulsion

Objective: This experiment investigates the antibacterial mechanism of genistein through antibacterial assays and network pharmacology,and develops novel formulations for genistein to select optimal formulations to enhance oral absorption,solubility and bioavailability of the drug.This study includes the investigation of the antibacterial activity of genistein,network pharmacology and molecular docking,and the preparation and characterization of microemulsions,which provide some clues for clinical trial studies.Methods: The antibacterial activity of genistein against Escherichia coli and Staphylococcus aureus was investigated based on antibacterial assays,and the MIC values were determined based on the multiplicative dilution method.Network pharmacology and molecular docking techniques were used to predict the key targets and related pathways for the antibacterial properties of genistein.The solubility of genistein in each oil phase,surfactant and co-surfactant was determined by high performance liquid chromatography,and the oil phase,surfactant and co-surfactant with higher solubility were selected,and the amount of oil phase,Km and distilled water were titrated by water droplet method to construct a pseudo-ternary phase diagram.The microemulsions were characterized by stability,transmittance,p H,particle size,PDI coefficient,conductivity and morphology of the microemulsions for the characterization experiments.Results: The results of antibacterial assay showed that genistein had good inhibitory activity against Staphylococcus aureus with a minimum inhibitory concentration of100 μM,and poor antibacterial activity against Escherichia coli.The results of network pharmacology prediction showed that the key targets in genistein to exert antibacterial effects might be ESR1,EGFR,MMP9,PTGS2,MMP2,APP,etc.GO functional enrichment analysis and KEGG pathway enrichment analysis screened5-hydroxytryptaminergic synapse,endocrine resistance,ABC transporter protein and other pathways to exert antibacterial effects,and the molecular docking results showed that by the molecular docking results showed that the top three targets of Degree value were ESR1,EGFR and MMP9 with good binding ability by Cytoscape’s plug-in Centiscape2.2 screening.By solubility results,the oil phase(polyethylene glycol glycerides of oleic acid),surfactants(Tween 80,polyethylene glycol glycerides of caprylic acid),and co-surfactants(anhydrous ethanol,1,2 propylene glycol,PEG-400)were selected.Three microemulsion formulations(ME-1,ME-2,ME-3)were selected by the results of pseudo-ternary phase diagram,and the maximum drug loading of genistein were 8.62±0.15 mg/m L,8.41±0.22 mg/m L,and 8.24±0.19mg/m L,respectively,and the particle sizes of microemulsions were between 24-36 nm.The screened microemulsions were identified by freeze-thaw cycles and high-speed centrifugation to possess stability,and their p H.The microemulsions screened by freeze-thaw cycles and high-speed centrifugation were found to be stable and their p H,PDI coefficient,conductivity,light transmission and morphology were all in accordance with the requirements of microemulsions.Conclusion: By investigating the antibacterial properties of genistein and predicting the potential targets and related pathways,we can provide a scientific basis and reference for the study of the antibacterial action mechanism of genistein.The preparation and characterization of the microemulsion of genistein led to the development of a new formulation of solubilized genistein that is both safe and stable.