Hydroquinone compound with anti-HCV (Hepatitis C Virus) activity and preparation method thereof
A hydroquinone and compound technology, applied in the field of hydroquinone compounds and their preparation, new compounds, can solve the problem that the anti-HCV activity of compound euchroquinolA has not been reported yet, and achieve simple and easy separation methods and high quality extracts Good, high yield effect
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Embodiment 1
[0024] B. Concentrate the extract in step A under reduced pressure as usual, recover ethanol until there is no ethanol smell, then use petroleum ether to extract repeatedly at room temperature according to the volume ratio of extract:petroleum ether=1:1 Second, after recovering the petroleum ether, the extract was obtained;
[0025] C. According to the volume ratio of extract: ethyl acetate=1:1, the extract of step B was repeatedly extracted 3 times with ethyl acetate at room temperature, and concentrated under reduced pressure to recover ethyl acetate to obtain 652 g of extract;
[0026] D. Put the extract from step C on a silica gel column (silica gel for mixing: 650 g, 80-100 mesh, silica gel for chromatography: 3 kg, 200-300 mesh) for chromatographic separation, first elute with chloroform, and then use Chloroform: methanol = 40:1, 10:1, 8:2, 6:4 volume ratio eluent gradient elution, and finally eluted with methanol to obtain five poles a, b, c, d, e, f Crude isolates of ...
Embodiment 2
[0036] The hydroquinone compound euchroquinol A was dissolved in DMSO, 0.22 mu M membrane filter sterilization, the concentration of the original drug solution was 10 mg / mL.
[0037] Human liver cancer cell line Huh 7.5.1 was treated with DMEM (Dulbecco’s modified Eagle’s medium, DMEM) containing 15% newborn bovine serum at 37°C and 5% CO 2 cultured in an incubator. Virus J6 / JFH1 was cultured from Huh 1 cells and stored at -80°C.
[0038] 1) MTT method to detect the cytotoxicity of samples
[0039] Huh 7.5.1 cells in the logarithmic growth phase were taken, and 9×10 3 cells / well cells were spread in 96-well plate, after 5 hours of attachment, add 2 mu L DMSO gradient dilution of the drug, 5-fold dilution, 5 dilutions, three replicate wells for each gradient, and set blank control (medium only), cell control, drug color control, DMSO control and anti-HCV positive drugs Ribavirin control, final volume 200 mu L / well. Place the culture plate at 37°C 5% CO 2 incubator f...
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