Application of Montelukast sodium in inhibiting infection of Zika virus, dengue virus and yellow fever virus
A technology of yellow fever virus and montelukast sodium, which is applied in the field of biotechnology and biomedicine, and can solve problems such as drugs that have not yet been developed with specific effects
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Embodiment 1
[0054] Example 1. Inhibition of ZIKV, DENV and YFV infection by montelukast sodium in vitro
[0055] The experimental operation of montelukast sodium inhibition of ZIKV, DENV and YFV infection can refer to: Zhiwu Sun, et al., Intranasal Administration of Maleic Anhydride-Modified Human SerumAlbumin for Pre-Exposure Prophylaxis of Respiratory Syncytial VirusInfection; Viruses 2015,7,798-819 ;doi:10.3390 / v7020798, as follows:
[0056] (1) ZIKV, DENV and YFV were amplified by inoculating C6 / 36 cells respectively (refer to SciChina LifeSci, doi: 10.1007 / s11427-016-5043-4).
[0057] (2) Use serum-free DMEM to serially dilute Montelukast sodium in a 96-well plate to the concentrations of 80 μM, 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM, and 0.625 μM, respectively. 50 μL, 3 replicates for each concentration.
[0058] (3) Dilute ZIKV (strains SZ01, MR766, FLR), DENV-2 or YFV-17D with serum-free DMEM so that the final concentration of the virus is 100 TCID50 (half the tissue culture...
Embodiment 2
[0068] Example 2. Inhibition of HCoV-OC43 and HCoV-229E infection by montelukast sodium in vitro
[0069] (1) The human coronavirus strains HCoV-OC43 (100TCID50) and HCoV-229E (500TCID50) were respectively mixed with the serially diluted montelukast sodium prepared in Example 1, and the resulting mixture was placed in 37 Incubate at °C for 30 minutes.
[0070] (2) The mixture of HCoV-OC43 and montelukast sodium was added to a 96-well plate with a monolayer of HCT-8 cells, and the mixture of HCoV-229E and montelukast sodium was added to a monolayer of Huh7 cells. In a 96-well plate, 3 replicates were set for each concentration. The 96-well plate was incubated at 33°C (HCoV-OC43) or 32°C (HCoV-229E) for 2 hours, after which the medium was replaced with fresh medium.
[0071] (3) When the cells in the positive group showed obvious CPE (cytopathic effect), CCK8 was used to detect the inhibitory activity of montelukast sodium on the virus. For the specific operation of CCK8 detec...
Embodiment 3
[0073] Embodiment 3 Toxicity detection of montelukast sodium on BHK21 cells
[0074] The toxicity detection operation of montelukast sodium on BHK21 cells can refer to: Zhiwu Sun, et al., Intranasal Administration of Maleic Anhydride-Modified Human Serum Albumin for Pre-Exposure Prophylaxis of Respiratory Syncytial Virus Infection. Viruses2015,7,798-819; doi :10.3390 / v7020798), as follows:
[0075] (1) Dilute montelukast sodium with serum-free DMEM to the concentrations of 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, and 0.3125 μM, respectively. The dosage volume per well is 100 μL, and each concentration is 100 μL. Set 2 repetitions.
[0076] (2) BHK21 cells were digested with EDTA-EGTA digestion solution, and the cells were diluted to 4*10 with DMEM containing 2% FBS 5 / mL, and added 100 μL / well to the 96-well plate in (1). Then at 37°C, 5% CO 2 cultured for 4 days.
[0077] (3) Carefully aspirate and discard the medium in the 96-well plate, add fresh DMEM cont...
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