Tubulin/VEGFR-2-IN-2 is an orally active dual inhibitor of tubulin and VEGFR-2, with IC50 values of 3.27 μM and 0.09 μM, respectively. This compound exerts antitumor effects through multiple pathways, including enhancing reactive oxygen species (ROS) production, disrupting mitochondrial membrane potential, inducing apoptosis, and causing cell cycle arrest. Tubulin/VEGFR-2-IN-2 also demonstrates significant anti-angiogenic properties in vitro, effectively inhibiting endothelial cell migration, invasion, and tube formation. Furthermore, in vivo, it inhibits angiogenesis, tumor growth, and metastasis. Tubulin/VEGFR-2-IN-2 is applicable for research on non-small cell lung cancer, breast cancer, gastric cancer, and lymphoma.

VEGFR2:0.09 μM

Tubulin/VEGFR-2-IN-2 (compound 19d) exhibits outstanding antiproliferative activity across various cancer cell lines, with IC 50 values of 0.26, 0.19, 0.04, 0.04, 12.06, and 4.70 μM for A549, MCF-7, MGC-803, U937, HEK 293 T, and RAW 264.7 cells, respectively. In MGC-803 cells, Tubulin/VEGFR-2-IN-2 (0-20 nM, 24 hours) demonstrates potent antiproliferative effects. At concentrations of 25-100 nM for 48 hours, this compound inhibits cell proliferation by disrupting tubulin polymerization in MGC-803 cells. Furthermore, it induces apoptosis through the activation of caspase proteins, reduces mitochondrial membrane potential in a dose-dependent manner, causes significant DNA damage and oxidative stress, leads to G2/M phase cell cycle arrest, and diminishes the migratory and invasive capabilities of MGC-803 cells. Additionally, Tubulin/VEGFR-2-IN-2 (20-80 nM, 6-8 hours or 48 hours) inhibits angiogenesis by suppressing the VEGFR-2 driven PI3K/AKT/MAPK pathway in human umbilical vein endothelial cells (HUVEC), resulting in the disruption of angiogenic structures, breakdown of tubular networks, and reduction in node numbers, major junctions, segment length, total branch length, grids, and average density. It also inhibits the migration and invasion of endothelial cells in HUVECs.

Tubulin/VEGFR-2-IN-2 (compound 19d) at a concentration of 100-400 nM in 1 mL water, administered for 18 hours, can inhibit angiogenesis in Tg(flk: EGFP) zebrafish embryos. Additionally, Tubulin/VEGFR-2-IN-2 at the same concentration and for 72 hours significantly suppresses tumor growth and metastasis in the bodies of Tg(flk: EGFP) zebrafish larvae induced by fluorescently labeled MGC-803 cells in a dose-dependent manner.