Synthesis and Anti‐Inflammatory Activity Evaluation of Benzoxazole Derivatives as New Myeloid Differentiation Protein 2 Inhibitors

Myeloid differentiation protein 2 (MD2), a key TLR4 adaptor protein for sensing LPS, plays an important role in inflammatory process and has been identified as a promising target for the treatment of a variety of inflammatory diseases. In our study, a series of benzoxazolone derivatives were synthesized, characterized and tested for anti‐inflammatory activity in vitro . The compounds 3c , 3d and 3g demonstrated the greatest anti‐inflammatory activity against IL‐6 with IC 50 values of 10.14±0.08, 5.43±0.51 and 5.09±0.88 μM, respectively. Furthermore, the bis‐ANS displacement assay revealed that these compounds competitively inhibited the binding between the probe bis‐ANS and the MD2 protein. The most active compound 3g , revealed a directly bind with MD2 protein via Arg90 binding and a dissociation constant value of 1.52×10 −6 mol L −1 as determined by the biological layer interference (BLI) assay. Our finding suggested that compounds 3g could be a promising lead compound as MD2 inhibitor for further anti‐inflammatory agent development.