A composition and combined medication method for treating enterovirus infection
A technology of enterovirus and composition, applied in the field of biomedicine
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Embodiment 1
[0136] Embodiment 1 single-component activity assay
[0137] Itraconazole, rupintravir, favipiravir, suramin and GW5074 can effectively inhibit the infection of cells by EV71 in a dose-dependent manner.
[0138] Figure 1. Itraconazole, rupinetravir, favipiravir, suramin and GW5074 inhibit the infection of EV71. (A) 2-fold dilutions of itraconazole, rufentravir, favipiravir, suramin and GW5074 were added to RD cells, and the cell viability was detected by CellTiter-Glo kit after adding virus or culture medium for 96 hours , to detect the inhibitory effect of four compounds on EV71 and their cytotoxicity. (B) Add virus and 2-fold dilutions of itraconazole, rupintravir, favipiravir, suramin and GW5074 to RD cells, collect supernatant after 48 hours of culture, and use TCID 50 method to determine the virus titer. Results were processed using Graphpad Pri sm5. The data in the figure are from two independent parallel experiments, and the error bars represent the standard deviati...
Embodiment 2
[0139] Example 2 Favipiravir acts on enterovirus 71 type 3D protein
[0140] By screening viruses resistant to favipiravir, the inventors obtained two strains of resistant viruses. The virus titer reduction experiment showed that both strains of the virus could produce drug resistance to favipiravir, and through sequence analysis, the inventors found that the same mutation appeared on the 3D protein. Drug-resistant virus phenotype experiments proved that EV71 with a mutation at position 121 of the 3D protein was resistant to favipiravir. Using the reverse genetics system, the inventors constructed a virus with the mutation, and verified its drug resistance to Favipiravir. The results showed that mutation of the 121st position of the 3D protein of enterovirus 71 from serine to aspartic acid would confer drug resistance to EV71, so the inventors concluded that Favipiravir acts on the 3D protein of EV71. In addition, the inventors tested the inhibitory effects of itraconazole, ...
Embodiment 3
[0142] Embodiment 3 combined activity test
[0143] The combination of itraconazole, rupintravir, favipiravir, suramin and GW5074 produced synergistic, additive or antagonistic effects in the treatment of EV71 infection. The result is as image 3 shown.
[0144] image 3 . The interaction effect of different drug combinations in the treatment of EV71 infection. The 3D graph was made using MacSynergy II software, and the data in the graph came from at least three independent parallel experiments. (A) combination of itraconazole and GW5074, (B) combination of itraconazole and suramin, (C) combination of suramin and rupinetravir, (D) combination of favipiravir and rupinetravir, (E ) combination of favipiravir and suramin, (F) combination of favipiravir and itraconazole. The horizontal plane represents the interaction of the two drugs as an additive effect, the point above the horizontal plane represents the effect of the two drugs as a synergistic effect, and the points belo...
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