Targeting innate immunity for antiviral therapy through small molecule agonists of the RLR pathway

To better understand the small molecule agonists that can promote IRF3 activation to induce innate immune gene expression could serve as novel antiviral compounds, we used microarray analysis to assess the transcriptional profiles driven by these compounds and to compare them to known activators of the intracellular innate immune response. We are particularly interested in knowing how the analogs derived by medicinal chemistry (KIN1408 and KIN1409) differ in signaling and gene expression profiles in THP-1 cells as compared to the parent KIN1400 and whether such differences may track with different biological outcomes. THP-1 cells were differentiated in 40nM PMA for 30 hrs and treated with the following controls for 20 hrs in cRPMI supplemented with 0.5% (v/v) DMSO: 0.5% DMSO alone, SeV (25 or 100 HAU/mL), IFNβ (25 or 100 IU/mL) or LPS (0.25 or 0.025 µg/mL). Alternatively, cells were treated in the same manner with the compounds KIN1400, KIN1408, or KIN1409 at doses of 10 µg/mL, 2.5 µg/mL, and 0.625 µg/mL. As an additional control, another set of cells were transfected with 2 µg/mL HCV pU/UC or XRNA in cRPMI without DMSO.