Application of phosphodiesterase inhibitor or pharmaceutical composition thereof in preparation of medicines for treating novel coronavirus pneumonia
A phosphodiesterase, coronavirus technology, applied in the field of medicine
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Embodiment 1
[0044] In this example, a preliminary evaluation of the effect of zadaverine on the novel coronavirus is carried out as follows:
[0045] (1) Use the following methods to study the impact of the novel coronavirus on the transcriptome of host cell lines:
[0046] 1. Experimental operation and analysis process
[0047] (1) In vitro experiment of viral infection of Huh7 (human liver cancer cells).
[0048] Huh-7 cells use DMEM+10% FBS medium, 37°C, 5% CO 2 standard environment. The cell suspension digested with trypsin was plated on a 12-well plate and cultured overnight. Discard the cell culture medium in the well the next day and replace it with a virus-containing medium (0.05 MOI). After incubation for 1 hour, discard the virus-containing medium, rinse with PBS three times, and replace with a standard medium. Lysed cells were harvested with TRIZOL at 3, 18, and 24 hours after infection. Transcriptome sequencing and analysis of Huh7 cells after virus infection.
[0049] (...
Embodiment 2
[0067] In this example, the in vitro inhibitory effect of zadavirin on the activity of the new coronavirus was determined by a cell-level experiment. And select other 3 phosphodiesterase inhibitors to verify the anti-new coronavirus activity.
[0068] 1. Experimental operation and analysis process
[0069] Six treatment concentrations (100 μM, 33.3 μM, 11.1 μM, 3.7 μM, 1.23 μM, 0.41 μM) were set for each drug, and a non-infected cell control and an infection control without drug treatment were set on each plate. Huh-7 cells use DMEM+10% FBS medium, 37°C, 5% CO 2 standard environment. The cell suspension after trypsinization (Huh7 cells were inoculated at 2.1×104 / 100 μL) was spread on a 12-well plate and cultured overnight. Discard the cell culture medium in the well the next day and replace it with virus-containing medium (0.05 multiplicity of infection). After incubation for 1 hour, discard the virus-containing medium, rinse with PBS three times, and replace with standard ...
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