Application of Mitoquinone in preparation of anti-coronavirus infection medicine
A technology of coronavirus and medicine, applied in the field of application in the preparation of anti-coronavirus infection medicine
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Embodiment 1
[0031] Time-dependent fluorescence intensity of substrate peptides metabolized by 3C-like proteases under different inhibitor concentrations
[0032] The specific implementation process is:
[0033] 1) The SARS-Cov-2 3C-like protease and substrate peptide stock solutions are stored in a -80°C refrigerator;
[0034] 2) Thaw the SARS-Cov-2 3C-like protease in a cryopreservation plate (-4 to 4°C) at room temperature, take 1uL and dilute it in 98uL borate borax buffer (PH=7.4), and add it to the detection plate;
[0035] 3) Add 1uL of different concentrations of inhibitors (Mitoquinone, concentrations of 0, 0.05mM, 0.1mM, 0.25mM, 0.5mM, 1mM, 2.5mM) into the solution obtained in the above step (2);
[0036] 4) Add 1uL of the same concentration of substrate peptide (0.5mM) to the solution obtained in the above step (3), incubate at 37°C with a fluorescent microplate reader, and monitor the fluorescence emission at 342nm excitation and 496nm with a fluorescent microplate reader , d...
Embodiment 2
[0039] Determination of Compound Mitoquinone's Inhibitory Ability to SARS-Cov-2 3C-like Protease
[0040] The specific implementation process is as follows:
[0041] 1) The SARS-Cov-2 3C-like protease and substrate peptide stock solutions are stored in a -80°C refrigerator;
[0042] 2) Thaw the SARS-Cov-2 3C-like protease in a cryopreservation plate (-4 to 4°C) at room temperature, take 1uL and dilute it in 98uL borate borax buffer (PH=7.4), and add it to the detection plate;
[0043] 3) Add 1uL of different concentrations of inhibitors (Mitoquinone, concentrations of 0, 0.05mM, 0.1mM, 0.25mM, 0.5mM, 1mM, 2.5mM) into the solution obtained in the above step (2);
[0044]4) Add 1uL of the same concentration of substrate peptide (0.5mM) to the solution obtained in the above step (3), incubate at 37°C with a fluorescent microplate reader, and monitor the fluorescence emission at 342nm excitation and 496nm with a fluorescent microplate reader , detect while incubating, incubate f...
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