Tubulin polymerization-IN-84 can inhibit the polymerization process of tubulin (Tubulin) by targeting the colchicine-binding pocket, with an IC50 value of 10.9 μM. Tubulin polymerization-IN-84 exhibits antiproliferative activity against four cell lines, namely Jurkat, B16-F10, HCT116 and MDA-MB-231, with corresponding IC50 values of 60 nM, 380 nM, 138 nM and 1.054 μM, respectively. In B16-F10 cells, Tubulin polymerization-IN-84 can induce G2/M phase cell cycle arrest and further trigger cell apoptosis. In addition, in the B16-F10 melanoma model, it can effectively inhibit the growth of tumor tissue; when combined with PD-L1 monoclonal antibody, it can also enhance the in vivo anti-tumor immune response, and can be used in the related research fields of T-cell acute lymphoblastic leukemia, melanoma, colon cancer and breast cancer.

MDA-MB-231 cells:1.054 μM, Tubulin polymerization:10.9 μM, Jurkat cells:60 nM, HCT116 cells:138 nM

Tubulin polymerization-IN-84 (Compound 5b) exhibits inhibitory effects on the proliferation of four cell lines, namely Jurkat, B16-F10, HCT116, and MDA-MB-231, with the corresponding IC50 values of 60 nM, 380 nM, 138 nM, and 1.054 μM, respectively [1].
When B16-F10 cells are treated with 0.5-5 μM Tubulin polymerization-IN-84 for 48 hours, it can induce G2/M phase cell cycle arrest. Among them, the proportion of G2/M phase cells in the control group is 9.75%, and that in the 5 μM dose group can reach 81.1% [1].
Likewise, at a concentration of 0.5-5 μM and a treatment duration of 48 hours, Tubulin polymerization-IN-84 can effectively induce apoptosis in B16-F10 cells [1].
Tubulin polymerization-IN-84 can directly bind to β-tubulin, with a KD value of 31.84 μM for the binding between the two [1].
In the concentration range of 0.1-5 μM, Tubulin polymerization-IN-84 can compete with EBI for binding to the colchicine-binding site on β-tubulin in B16-F10 cells in a concentration-dependent manner [1].
Treating B16-F10 cells with 0.5-5 μM Tubulin polymerization-IN-84 for 12 hours can lead to the depolymerization of the microtubule network in the cytoplasm of the cells [1].
When HUVEC cells are treated at a concentration of 0.5-5 μM for 6 hours, Tubulin polymerization-IN-84 can dose-dependently disrupt the capillary-like network structure formed by HUVEC cells on Matrigel matrix and reduce the number of network connection points [1].

When Tubulin polymerization-IN-84 is administered by intraperitoneal injection at a dose of 10 mg/kg, once daily for 14 consecutive days, it can effectively inhibit the growth of B16-F10 melanoma tumors in C57BL/6 mice, with a tumor growth inhibition rate (TGI) of 27.5% [1]. When Tubulin polymerization-IN-84 is co-administered with PD-L1 monoclonal antibody (both at a dose of 10 mg/kg, total dose 10 + 10 mg/kg), its anti-tumor efficacy is further enhanced, with the tumor growth inhibition rate (TGI) increased to 36.2%, and no significant weight loss was observed in the mice during the entire administration period [1].