Application of Onalspib in preparation of medicine for preventing and/or treating adenovirus infection
An adenovirus and drug technology, applied in the field of medicine, can solve problems such as no adenovirus infection found, and achieve the expected effects of good safety, good safety, and short medication cycle
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Embodiment 1
[0039] Example 1: Evaluation of cytotoxicity of Onalespib in Vero cell line
[0040] 1. Cell culture
[0041] After 2 passages of cryopreserved and resuscitated cells, use DMEM medium containing 10% fetal bovine serum and double antibody (penicillin 100U / mL, streptomycin 100μg / mL) for expansion and culture, and the seeding density is not less than 1 ×10 4 cell / mL, passage density not higher than 5×10 4 cells / mL.
[0042] 2. Drug treatment of cells
[0043] Vero cells by 1 x 10 4 Cells / well (volume 100 μL) were seeded in 96-well cell culture plates, and cultured for 24 hours until the confluence of cell wells reached 80%; 200 μL of culture medium (DMEM medium + 2% serum + double antibody) was used in each well to prepare drugs, and Add to the corresponding cell wells and mix. Seven drug concentration gradients were set in the drug treatment group, and each gradient concentration was set up with 2 duplicate wells, and the final concentrations were 0.007 μM, 0.02 μM, 0.06 μ...
Embodiment 2
[0048] Example 2: Evaluation of anti-AdV3 adenovirus activity of Onalespib in Vero cell line
[0049] 1. Cell culture
[0050] After 2 passages of cryopreserved and resuscitated cells, use DMEM medium containing 10% fetal bovine serum and double antibody (penicillin 100U / mL, streptomycin 100μg / mL) for expansion and culture, and the seeding density is not less than 1 ×10 4 cell / mL, passage density not higher than 5×10 4 cells / mL.
[0051] 2. Drug treatment of cells
[0052] Vero cells by 1 x 10 4 Cells / well (volume 100 μL) were inoculated into 96-well cell culture plates, and cultured for 24 hours until the confluence of cell wells reached 80%; AdV3 virus of 0.55MOI (multiplicity of infection) was added to both the drug-treated group and the virus control group, while the drug-treated group Add each gradient concentration of the drug (with 5 μM as the starting concentration, 7 gradients of serial 3-fold serial dilution, two duplicate wells for each gradient) to a total vol...
Embodiment 3
[0062] Example 3: Evaluation of anti-AdV5 adenovirus activity of Onalespib in Vero cell line
[0063] 1. Cell culture
[0064] After 2 passages of cryopreserved and resuscitated cells, use DMEM medium containing 10% fetal bovine serum and double antibody (penicillin 100U / mL, streptomycin 100μg / mL) for expansion and culture, and the seeding density is not less than 1 ×10 4 cell / mL, passage density not higher than 5×10 4 cells / mL.
[0065] 2. Drug treatment of cells
[0066] Vero cells by 1 x 10 4 Cells / well (volume 100 μL) were inoculated in 96-well cell culture plates, and cultured for 24 hours until the confluence of cell wells reached 80%; 1.1 MOI (multiplicity of infection) AdV5 virus was added to the drug treatment group and the virus control group, while the drug treatment group Add each gradient concentration of the drug (with 5 μM as the starting concentration, 7 gradients of serial 3-fold serial dilution, two duplicate wells for each gradient) to a total volume of...
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