Application of EGCG palmitate to drugs for treating or preventing hepatitis C virus infection
A technology of hepatitis C virus and palmitate, which is applied in the direction of antiviral agents, can solve problems such as the treatment effect of hepatitis C virus that is not mentioned, and achieve the effects of reducing replication efficiency, stabilizing results, and improving preventive effects
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Embodiment 1
[0038] Embodiment 1: the toxicity test of EGCG palmitate to host cell,
[0039] When the HuH7 cells cultured for 24-48 hours are almost covered with a monolayer, discard the cell culture medium, add trypsin to digest, transfer to a 96-well sterile cell culture plate, 100 μl per well (1*10 5cell / ml, the same below). Place it in a cell culture incubator for 18-24 hours to make the cells grow into a single layer for later use. Dilute the EGCG palmitate stock solution (DMSO configuration, mass volume ratio 50mg / mL) with the cell maintenance solution to make five concentration gradients, and then add different concentrations of drugs to the cell culture in which the cell culture solution was discarded In the wells, 100 μl per well, repeat 3 wells for each concentration, and set up cell control wells (no drug addition, 99uL culture solution + 1uL DMSO), blank wells (no cells, only add cell culture solution), set at 37 ° C, cell culture In the box, after culturing for 48 hours, add...
Embodiment 2
[0044] Embodiment 2: HCV is to the mensuration of cell half infection dose (TCID50)
[0045] Transfer the HuH7 cells cultured into a monolayer to a 96-well cell culture plate, and culture them in a cell culture incubator for 18-24 hours. The virus stock solution was serially diluted 10 times with cell maintenance solution (10 -1 -10 -8 ). Discard the culture medium in each well of HuH7 cultured into a single layer, wash each well with PBS 3 times, add 100 μl of virus dilution solution of different concentrations to each well, absorb at 37°C for 1.5 h, discard the virus dilution solution, and add to each well 100 μl of maintenance solution, each concentration was repeated 3 times, and normal cell control wells (without adding virus) were set. The cytopathic changes (CPE) in each well were observed daily for 3 consecutive days, and the CPE conditions were recorded. Then calculate the cell half infection amount of the virus by the Reed-Muench method, and the converted virus t...
Embodiment 3
[0047] Antiviral Effect of EGCG Palmitate on HCV
[0048] 1) Preventive effect of EGCG palmitate on HCV virus
[0049] When the HuH7 cells cultivated for 24-48 hours are almost covered with a single layer, discard the cell culture medium, add trypsin to digest, and transfer to a 12-well sterile cell culture plate, 1 mL per well. Place it in a cell culture incubator for 18-24 hours to make the cells grow into a single layer for later use. Dilute the EGCG palmitate stock solution with the cell maintenance solution into five concentration gradients required for the experiment. After dilution, add different concentrations of the drug into the cell culture wells where the culture solution was discarded, 1mL per well, each concentration Repeat 3 wells, and set up antiviral positive control cell wells (add virus, add IFN-α1 to 1000U / mL, add culture medium), virus control wells (do not add drugs, add virus, add cell maintenance solution) and cell control wells (No virus, no medicine...
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