Figure 4

The inhibition potential of RC-1 against the activity of DENV-2 NS2B-NS3pro. End point protease inhibition assay was carried out to evaluate the inhibitory potential of RC-1 against the protease’s ability to cleave a fluorogenic peptide substrate (Boc-Gly-Arg-Arg-AMC). The reaction mixtures consisted of 20 μM substrate, 2 μM of enzyme, with or without RC-1 of varying concentrations incubated at 28°C, 37°C or 40°C for 30 minutes. The results were normalized with the substrate alone and substrate with protease incubated at same temperatures. The inhibition was found to be temperature dependent with the highest concentration (120 μM) at 28°C and the lowest concentration (37.5 μM ) at 40°C.