Discovery ofc‑Kit as a New Therapeutic Targetin LPS-Induced Acute Lung Injury through Novel Phenylamide DerivativeD9

Discovering effective anti-inflammatory drugs and targets is a critical research priority. Herein, 28 novel phenylamide derivatives were designed and synthesized. Compound <b>D9</b> showed favorable anti-inflammatory activities in acute lung injury (ALI) and sepsis mouse models. The target of <b>D9</b> was predicted and studied, and c-Kit was identified by surface plasmon resonance (SPR) and cellular thermal shift assay (CETSA), which demonstrated high selectivity in kinase activity profiling. It was further verified that c-Kit participated in the LPS-induced inflammatory response in J774A.1, RAW264.7, MPMs, and lung tissue. Additionally, c-Kit is proved to be essential for LPS-induced activation of the NF-κB pathway. Finally, c-Kit was confirmed as a therapeutic target in the LPS-induced ALI with c-Kit gene knockdown and was further verified as the target of <b>D9</b>. Notably, there has been no report of c-Kit’s influence on the LPS-induced inflammatory response. Therefore, c-Kit was identified as a new therapeutic target for the LPS-induced ALI.