Aurora A-IN-5 is a potent and highly selective Aurora A inhibitor (IC50= 0.02 μM) with 362 times higher selectivity for Aurora A over Aurora B. This selectivity is due to its unique C−H/π interactions, enhanced hydrophobic contacts, open binding pocket, and tighter protein stacking. Aurora A-IN-5 inhibits Aurora A autophosphorylation, thereby suppressing cancer cell proliferation through G2/M phase arrest, triggering apoptosis, and inhibiting colony formation. It also inhibits tumor growth in MDA-MB-231 xenograft mouse models. Aurora A-IN-5 is applicable for research related to breast cancer, cervical cancer, prostate cancer, and lymphoma.

Aurora A:0.02 μM

Aurora A-IN-5 (compound 5h) exhibits potent antiproliferative activity against various cancer cell lines with IC50 values of K562 (2.87 μM), MCF-7 (1.50 μM), HeLa (0.85 μM), DU145 (1.85 μM), MOLT-4 (2.25 μM), and MDA-MB-231 (2.04 μM). When applied at 0.5 μM for 48 hours, this compound induces spindle abnormalities and chromosome misalignment in MDA-MB-231 cells. At concentrations of 2.5-10 μM for 24 hours, it effectively inhibits cell division in MDA-MB-231 cells by arresting the G2/M phase of the cell cycle. Aurora A-IN-5 triggers apoptosis in MDA-MB-231 cells in a dose-dependent manner at 2-20 μM over 24 hours. Furthermore, at 2 μM over 14 days, it significantly inhibits clonogenic formation of MDA-MB-231 cells, thereby suppressing proliferation. The compound demonstrates strong, selective, dose-dependent inhibition of Aurora A phosphorylation at Thr288 from 0-1 μM, and significantly stabilizes the Aurora A protein in MDA-MB-231 cells through cellular thermal shift analysis (46-62 °C), confirming direct target binding. Additionally, Aurora A-IN-5 is toxic to mouse embryonic osteoblast MC3T3-E1 cells (IC50 = 7.135 μM) but shows minimal toxicity towards normal liver LO2 cells (IC50 = 43.86 μM).

Aurora A-IN-5 (30 mg/kg, administered intraperitoneally every other day for 20 days) effectively inhibits tumor growth in MDA-MB-231 xenograft mouse models.