Discovery and Characterization of 2-Aminobenzimidazole Derivatives as Selective NOD1 Inhibitors

NLR family proteins play important roles in innate immune response. NOD1 (NLRC1) activates various signaling pathways including NF-kappa B in response to bacterial ligands. Hereditary polymorphisms in the NOD1 gene are associated with asthma, inflammatory bowel disease, and other disorders. Using a high throughput screening (HTS) assay measuring NOD1-induced NF-kappa B reporter gene activity, followed by multiple downstream counter screens that eliminated compounds impacting other NF-kappa B effectors, 2-aminobenzimidazole compounds were identified that selectively inhibit NOD1. Mechanistic studies of a prototypical compound, Nodinitib-1 (ML130; CID-1088438), suggest that these small molecules cause conformational changes of NOD1 in vitro and alter NOD1 subcellular targeting in cells. Altogether, this inaugural class of inhibitors provides chemical probes for interrogating mechanisms regulating NOD1 activity and tools for exploring the roles of NOD1 in various infectious and inflammatory diseases.