New antibacterial compounds
a technology of antibacterial compounds and compounds, applied in the field of new antibacterial compounds, can solve the problems of increasing the risk of infection spreading to others, ineffective standard treatment, and inability to meet the needs of patients, and achieve the effect of facilitating preparation and detection
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example 1
Inhibition of DNA Gyrase and Topo IV in E. coli and S. aureus
[0557]The above compounds were tested for the inhibition of the enzyme DNA gyrase in a gyrase supercoiling assay and for the inhibition of the enzyme topoisomerase IV in a decatenation assay, in both Gram positive and Gram negative bacteria, according to the following methods.
[0558]Both the assays were carried out according to a set-up method modified from the article to Blanche F, et al. “Differential Behaviors of Staphylococcus aureus and Escherichia coli Type II DNA Topoisomerases”, Antimicrob. Agents Chemother., 1996, Vol. 40, No. 12 p. 2714-2720.
[0559]The compounds were screened at single concentration (200, 100 or 50 μM), in duplicate.
[0560]Ciprofloxacin and novobiocin were used as reference compounds, at single concentration of 200 and 50 μM, respectively.
[0561]DNA Gyrase Supercoiling Assay.
[0562]Reagents from S. aureus and E. coli Gyrase Supercoiling Assay kits (Inspiralis, UK) were used. A master mix with a total...
example 2
Determination of IC50
[0586]The compounds that in the above example 1 showed an inhibitory activity were further assayed in concentration-response curve (eight half-log concentrations ranging from 0.1 to 300 μM) in order to determine the IC50.
[0587]The supercoiled or decatenated DNA bands obtained as described in Example 1 were analysed as follows.
[0588]Bands were analyzed by gel documentation equipment (Syngene, Cambridge, UK) and quantitated using Syngene Gene Tools software. Raw gel data (fluorescent band volumes) collected from Syngene, GeneTools gel analysis software were converted to a percentage of the 100% control (the fully supercoiled or decatenated DNA band). These data were analyzed using SigmaPlot Version 12.3 (2013). The IC50 data were calculated by using the global curve fit non-linear regression tool by selecting the Single, 2 Parameter fit function from the Exponential Decay equation category.
[0589]The results are reported in the following table 3.
TABLE 3CompoundIC5...
example 3
Determination of IC50
[0590]Compounds 301-303, 314, 343 and 354 were assayed in concentration-response curve (eight half-log concentrations ranging from 0.1 to 300 μM) in order to determine the IC50, following the procedure described in example 2.
[0591]The results are reported in the following table 4.
TABLE 4CompoundIC50IC50IC50IC50No.DNA gyraseTopo IVDNA gyraseTopo IV3010.460.430.7270.33020.590.120.583.263032.670.629.92>1003140.851.147.1>1003430.911.271.988.733540.470.180.350.92
[0592]The data of Table 3 are comparable to the data of Table 2, and confirmed the activity of Compounds 301-303, 314, 343 and 354 to effectively inhibit both DNA gyrase and Topo IV of E. coli and / or S. aureus.
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