Design, synthesis, and evaluation of novel benzoylhydrazone derivatives as Nur77 modulators with potent antitumor activity against hepatocellular carcinoma

Nur77 modulators have emerged as a promising therapeutic approach for hepatocellular carcinoma. In this study, a structure-based rational drug design approach was used to design and synthesise a series of 4-((8-hydroxy-2-methylquinolin-4-yl)amino)benzoylhydrazone derivatives based on the binding characteristics of our previously reported <b>10g</b> and the native ligand <b>3NB</b> at the binding Site C of Nur77. Cell-based cytotoxicity assays revealed that compound <b>TMHA37</b> demonstrated the highest cytotoxicity against all tested cancer cells. The induced fit docking and binding pose metadynamics simulation suggested that <b>TMHA37</b> was the most promising Nur77 binder at Site C. Molecular dynamics simulation validated the stable binding of <b>TMHA37</b> to Nur77’s Site C but not to Sites A or B. Specifically, <b>TMHA37</b> bound strongly to Nur77-LBD (<i>K</i>_D = 445.3 nM) and could activate Nur77’s transcriptional activity. Furthermore, <b>TMHA37</b> exhibited antitumor effects by blocking the cell cycle at G2/M phase and inducing cell apoptosis in a Nur77-dependent manner.