BAL-0028 is a potent inhibitor of NLRP3 signaling in multiple human cell types. (A) Structure of BAL-0028. (B–D) Comparison of BAL-0028 and MCC950 in IL-1β release assays from PMA-differentiated THP-1 cells stimulated with LPS and (B) nigericin, (C) ATP, or (D) MSU. (E–I) Comparison of BAL-0028 and MCC950 in IL-1β release assays from LPS and nigericin-stimulated human monocytes (E), iCell microglia (F), HMDM (G), iMacs (H), and human whole blood (I). (B–F and I) Graph symbols show average IL-1β values relative to vehicle control ± SEM from independent experiments performed in triplicate; the IC₅₀ curve was fitted by nonlinear regression analysis. (G and H) Graph symbols show average values relative to vehicle control from independent experiments (indicated by different symbols) performed in triplicate ± SEM. Compounds are shown in nanomolar (nM) concentrations. N = 73 for BAL-0028 and N = 29 for MCC950 (B), N = 2 (C, D, and E), N = 3 (F and H), N = 3 donors (G), and N = 4 donors (I).