Fig. 6: RPG-01-132 has consistent activity against the four DENV serotypes and the ST148-resistant DENV2 C S34L mutant.
a Orientations of two molecules of ST148 with the DENV2 C tetramer (PDB: 6VG5)41. The binding pocket of one ST148 molecule (shown in orange) is shown in detail. b Alignment of amino acid sequences of DENV1 WP74, DENV2 NGC and 16681, DENV3 THD3, and DENV4 TVP360. Five residues (30, 35, 37, 38, and 50) are variable, and two residues (F33 and F53) are conserved across all four serotypes. c–i Antiviral activity was measured against DENV1 WP74, DENV2 NGC, DENV3 THD3, DENV4 TVP360, DENV2 16681, and DENV2 16681 C-S34L strains following the experimental outline in Fig. 3a. c RPG-01-132 exhibits consistent, CRBN-dependent antiviral activity across all four DENV serotypes. Black and blue symbols represent treatment with DMSO/compound alone or co-treatment with DMSO/compound and lenalidomide, respectively. Each data point represents an independent biological replicate, with two technical replicates averaged per experiment. Bars represent means normalized to DMSO ± standard deviation from n = 2 independent biological replicates. No statistical hypothesis testing was performed for this panel. (d) DENV2 16681 is sensitive to ST148 (EC50 0.2 ± 0.05 µM), but the DENV2 16681 C-S34L mutant is not (EC50 could not be determined). (e) RPG-01-132 exhibits comparable antiviral activity against wildtype and ST148-resistant viruses (EC50 1 and 0.6 µM, respectively). f–i RPG-01-132 exhibits comparable degradation activity against DENV2 16681 and DENV2 16681 C-S34L viruses with DC50 values of 3 and 1.85 µM, respectively. Viral titers in c-e are presented as means normalized to DMSO ± standard deviation. Antiviral EC50 values were determined by nonlinear regression analysis of the data from n = 2 independent experiments. Band intensities in f and g were quantified by ImageJ. DC50 values were determined by nonlinear regression analysis of the mean capsid abundance normalized to DMSO from n = 2 independent experiments. Source data are provided as a Source Data file.
