Oxadiazole-inosine type compound as tyrrs inhibitor and its preparation method and use
The technology of a compound and oxadiazole is applied in the application field in the preparation of anti-infective drugs, and achieves the effects of high bacteriostatic activity, good inhibition and killing effects
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Embodiment 1
[0024] Example 1: 2-(((2R,3S,4R,5R)-3,4-dihydroxy-5-(6-oxo-1H-purin-9(6H)-yl)tetrahydrofuran-2-yl) Preparation of methylamino)-N-(5-phenyl-1,3,4-oxadiazol-2-yl)acetamide (1)
[0025] Step 1: Weigh 5.58g (0.05mol) semicarbazide hydrochloride and 8.20g (0.1mol) anhydrous sodium acetate in a flask, add 25mL water to dissolve, then dissolve 5.31g (0.05mol) benzaldehyde to 25mL water and ethanol, and then drop the ethanol solution of benzaldehyde into the flask. After the dropwise addition, react at room temperature for 1 hour. Purified by column chromatography, the volume ratio of the eluent is: petroleum ether:AcOEt=6:1, and 7.07g of white solid benzaldehyde semicarbazone was obtained, the yield: 87%, melting point: 155-157°C.
[0026] Step 2: Put 8.16g (0.05mol) of benzaldehyde semicarbazone and 8.20g (0.1mol) of anhydrous sodium acetate in a flask, add 55mL of glacial acetic acid to dissolve it, and add 2.56mL (0.05mol) of bromine after dissolution, After the dropwise additio...
Embodiment 2
[0040] Example 2: Extraction of TyrRS and determination of the activity of compounds on TyrRS
[0041] TyrRS from Staphylococcus aureus was expressed in E. coli and purified by Sephadex chromatography. The activity of TyrRS was determined by aminoacylation reaction. The enzyme reaction mixture consists of the following components: 100mM TrisHCl pH7.9, 50mMKCl, 16mMMgCl 2 , 5mMATP, 3mM dithiothreitol, 4mg / mL Escherichia coli MRE600tRNA and 10μM [3H]tyrosine (activity 1.48-2.22TBq / mmol). Mix and incubate TyrRS (0.2nM) and different concentrations of test substances at room temperature for 10 minutes, then add an equal amount of the above enzyme reaction mixture preheated to 37°C, and after co-incubating for 5 minutes, add an equal volume of 7% glacial trichloro Acetic acid solution was used to terminate the reaction, filtered through a 96-well Millipore filter plate, and the filtrate was detected by a scintillation counter, and each sample was repeated 4 times. The one withou...
Embodiment 3
[0042] Example 3: Antibacterial Activity of Compounds
[0043] Bacteria were suspended in MH medium at a concentration of approximately 10 5 cfu mL -1 , add the bacterial solution to a 96-well plate (100 μL of bacterial solution per well), use the culture medium as a blank control, replace the test substance with DMSO as a negative control, use penicillin G as a positive control for Gram-positive bacteria, and use penicillin G as a positive control for Gram-positive bacteria. Kanamycin was used as a positive control for Shi-negative bacteria, and ketoconazole was used as a positive control for fungi. Dissolve the test substance in DMSO to prepare 800, 400, 200, 100, 50, 25 μg·mL -1 solution (for MIC 50 Less than 5μg·mL -1 Yes, when carrying out one-step experiment, the prepared concentration gradient is 50, 25, 12.5, 6.25, 3.1, 1.5 μg·mL -1 ), added to a 96-well plate in an amount of 11 μL per well [the final concentrations of the drug solution were 80, 40, 20, 10, 5, 2.5...
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