HLM006474 is a pan inhibitor of E2F. This inhibits E2F4 DNA-binding (IC50: 29.8 µM in A375 cells).

E2F4 DNA-binding:29.8 μM

HLM006474 causes the viability of both SCLC and NSCLC lines (IC50 ranging from 15 to 75 μM). HLM006474 displays little activities against E2F4 DNA-binding in A375 cells at 10 and 20 μM, apparently inhibits E2F4 DNA-binding at 40 μM, and increasingly suppresses the effect at 60 and 80 μM concentrations. HLM006474 dramatically causes cyclin D3 protein expression and is a potent inducer of PARP cleavage[1]. HLM006474 (20 μM) weakly synergizes with paclitaxel, but there is antagonism between HLM006474 and cisplatin and gemcitabine in H1299 cells[2]. HLM006474 (40 μM) inhibits E2F4 activity via suppression of its DNA-binding activity and down regulation of its expression and HLM006474 (40 μM) also obviously causes apoptosis in A375 and 231 cell lines for 24 hours. HLM006474 (60 μM) enhances the expression of several known E2F-regulated genes after short treatments in H292 and H1299 cell lines. HLM006474 leads to a reduction in the mRNA levels of MAD2. HLM006474 apparently suppresses the increase of Mad2 protein and pRb-S780 signal but not the level of Skp2 protein in human lung cancer A549 cellss.