HLM006474 exhibits minimal activity against E2F4 DNA-binding in A375 cells at lower doses (10 and 20 µM), but clearly inhibits E2F4 DNA-binding at higher concentrations (40 µM and above), progressively increasing its suppressive effect at 60 and 80 µM. At 40 µM, HLM006474 hinders E2F4 activity by both blocking its DNA-binding capability and reducing its expression levels. This concentration also significantly triggers apoptosis in A375 and 231 cell lines within 24 hours, significantly cuts down cyclin D3 protein levels, and effectively induces PARP cleavage, demonstrating its potential as a potent apoptotic inducer^[1].

HLM006474 reduces the viability of both small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) lines, with IC50 values ranging between 15 and 75 µM. At 60 µM, it boosts the expression of several E2F-regulated genes after brief exposures in H292 and H1299 cell lines. Although HLM006474 (20 µM) exhibits slight synergistic effects with paclitaxel, it shows antagonistic interactions with cisplatin and gemcitabine in H1299 cells^[2].