Table 1.
Summary of NITD008 results.
| Virus ID | CPEa EC50 (μM) | CFIc EC50 (μM) | VTRd EC50 (μM) | CC50 (μm)a | SI (CPE) | SI (CFI) | SI (VTR) |
|---|---|---|---|---|---|---|---|
| AHFV | 3.31 ± 0.49 | 1.51 | 9.29 | >100 | >30 | >66 | >11 |
| KFDV | 1.42 ± 0.30 | 0.36 | 4.01 | >100 | >70 | >278 | >25 |
| OHFV | 0.61 ± 0.09 | 0.14 | 3.04 | >100 | >164 | >714 | >33 |
| TBEV | 0.90 ± 0.29 | 0.226 | 2.99 | >100 | >111 | >442 | >33 |
| EBOV-GFPb | >100 | ND | ND | >100 | ND | ND | ND |
Effective 50% inhibition and cytotoxic concentrations (EC50 & CC50) were calculated by non-linear regression analysis using GraphPad 6 software. Selectivity index (SI) = CC50/EC50. ND = Not determined.
Cytopathic Effect (CPE) assay. 4 independent experiments were performed for each virus in A549 cells. In each experiment, there were 4 biological replicates for each dilution of NITD008. ± indicates standard deviation from the mean values from the 4 experiments.
Ebolavirus replication was measured indirectly in Vero cells by measuring fluorescence levels emitted by the enhanced green fluorescent protein in a Biotek Synergy H1MD plate reader. One experiment was performed in which there were 4 biological replicates for each dilution of NITD008 treatment.
Cell-based flavivirus immunodetection (CFI) ELISA. One experiment was performed in A549 cells in which there were 3 biological replicates for each dilution of NITD008 against each virus. Chemiluminescence served as the indicator of viral antigen levels present, and was measured in a Biotek Synergy H1MD plate reader.
Virus titer reduction (VTR). One experiment was performed in A549 cells in which there were 4 biological replicates for each dilution of NITD008 against each virus.