Application of doxylamine succinate in preparing drug for treating or preventing influenza virus
A technology of doxylamine succinate and prevention of influenza, which is applied in the field of medicine to achieve the effect of broad-spectrum antiviral activity
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Embodiment 1
[0028] Embodiment 1: Evaluation of anti-influenza virus activity of doxylamine succinate
[0029] 1. Experimental materials and methods
[0030] 1.1 Cells, viruses and drugs
[0031] MDCK cells were purchased from ATCC, virus A / PR8 / 34H1N1 was amplified from chicken embryo culture, and doxylamine succinate was purchased from Sigma;
[0032] 1.2 Experimental Instruments
[0033] Multifunctional detector PerkinElmer, inverted microscope Costar
[0034] 2. experimental method:
[0035] 2.1 Cell culture:
[0036] 37°C, 5% (volume ratio, the same below) CO 2 cultured in a humidified incubator. A DMEM medium containing 10% (volume ratio, the same below) FBS, 100 U / mL of penicillin and streptomycin was used. Cells were subcultured after reaching 90% confluence, and the subculture ratio was 1 / 3–1 / 4.
[0037] 2.2 Virus culture:
[0038] Take 9-11 day-old SPF chicken embryos, check them with an egg tester before inoculating the virus, and mark them at a position far away from ...
Embodiment 2
[0053] Example 2: Broad-spectrum research on anti-influenza virus activity of doxylamine succinate
[0054] In this experiment, the antiviral activity of doxylamine succinate on influenza A virus subtype H1N1 strain, influenza A virus H3N2 strain and influenza B virus strain was detected.
[0055] 1. The virus strains used in the experiment include: A / human / Hubei / 1 / 2009 (H1N1), A / human / Hubei / 3 / 2005 (H3N2), Influenza B virus
[0056] 2. Divide MDCK cells into 2*10 4 Cells / well were seeded in 96-well cell culture plates, cultured in a 37°C cell culture incubator for 14-18 hours, and the cells were grown into a single layer before use. Discard the culture medium in the well plate, wash it twice with PBS, add 100TCID50 / well virus solution to infect the cells, and add drugs with different concentration gradients at the same time (with 500μM as the initial concentration, serially dilute 8 gradients by 2 times, each gradient 2 A total of 200 μl of maintenance medium (DMEM+0.3%BSA...
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