Identification and validation of a novel dual small-molecule TLR2/8 antagonist.

•Endosomal- and surface-located TLRs can be targeted by a small-molecule antagonist.•Modeling-guided synthesis approach identified compound 24 as dual TLR2/8 antagonist.•Compound 24 potently and selectively reduced TLR2- and TLR8-mediated signaling.•Compound 24 showed characteristics of non-competitive antagonism at TLR2.•Computational docking studies predicted a plausible binding mode at TLR2 and TLR8.