Tubulin polymerization-IN-6 (compound 5f) is a potent inhibitor of tubulin polymerization, with an IC50 of 1.09 μM. It not only inhibits cell migration and tube formation but also has anti-angiogenic properties. Additionally, Tubulin polymerization-IN-6 has been found to effectively hinder tumor growth in HT29 xenograft Balb/c nude mice [1].

Tubulin polymerization-IN-6 (compound 5f), across various concentrations and periods, demonstrates significant anti-cancer activities, as evidenced by its broad-spectrum anti-proliferative effects against several cancer cell lines (MCF-7, MDA-MB-231, A549, Hela, and HT29), with notable efficacy even against drug-resistant strains (MCF-7/TxR and A549/TxR). This compound inhibits colony formation by tumor cells, disrupts cell migration, and impedes angiogenesis in HUVEC cells. It achieves these effects, in part, by competing with colchicine for binding, thereby inhibiting tubulin polymerization. Moreover, Tubulin polymerization-IN-6 induces cell cycle arrest and apoptosis in HT29 cells by modulating relevant proteins, decreasing mitochondrial membrane potential, and increasing reactive oxygen species (ROS) production. Detailed analyses, including Western Blot and Immunofluorescence, reveal dose-dependent impacts on tubulin dynamics and on the regulation of proteins crucial for cell cycle progression and apoptosis, thereby highlighting the compound's potential as a multi-faceted anti-cancer agent.

Tubulin polymerization-IN-6, also referred to as compound 5f, demonstrated a dose-dependent inhibitory effect on tumor growth in HT29 xenograft Balb/c nude mice when administered intraperitoneally at dosages ranging from 0 to 10 mg/kg once every two days for three weeks, achieving a tumor weight reduction of 75.5% at the highest dosage. Additionally, a single intravenous dose of 10 mg/kg in SD rats revealed significantly improved pharmacokinetic properties, including an eight-fold increase in half-life and a two-fold enhancement in the area under the curve (AUC), indicating better systemic exposure and potential effectiveness of Tubulin polymerization-IN-6 against cancerous tumors.