Tubulin polymerization-IN-4 is an effective inhibitor of tubulin polymerization (IC50=4.6 μM）。 Tubulin polymerization-IN-4 can destroy tubulin polymerization, block cell cycle at G2/M phase, induce apoptosis, and inhibit cell cloning and migration. Tubulin polymerization-IN-4 can also damage the vascular system.

Tubulin polymerization-IN-4 (compound 9j) (0-1 μM; 48 hours) exhibits sub-micromolar inhibitory activities against HeLa, SiHa and MS751 [1]. Tubulin polymerization-IN-4 (3, 6 and 12.5 μM; 0-20 min) inhibits tubulin polymerization in a concentration-dependent manner with the inhibition percentages of 39%, 54%, and 77% at 3, 6 and 12.5 μM [1]. Tubulin polymerization-IN-4 (1-100 μM; 2 hours) inhibits the formation of EBI-β-tubulin adduct in a concentration-dependent manner [1]. Tubulin polymerization-IN-4 (0.2 μM; 1 and 2 hours) disrupts the HUVEC-formed vascular tube [1]. Tubulin polymerization-IN-4 (0.1-0.4 μM; 24 hours) increases cell distribution to the G2/M phase in a concentration-dependent manner [1]. Tubulin polymerization-IN-4 (0.1-0.4 μM; 24 hours) induces apoptosis of HeLa cells [1]. Tubulin polymerization-IN-4 (20, 50, 100 nM; 14 days) reduces new colony formation and suppresses HeLa cell growth for 14 days in a dose-dependent manner [1]. Tubulin polymerization-IN-4 (0.1, 0.2 and 0.4 μM; 24 hours) effectively inhibits the migration of HeLa cells in a concentration-dependent manner [1]. Tubulin polymerization-IN-4 (0-200 μM; 24 hours) exhibits good renal safety profile, with IC 50 of 188 ± 16 μM in HK-2 cells [1]. Cell Proliferation Assay Cell Line: HeLa, SiHa and MS751 [1] Concentration: 0-1 μM Incubation Time: 48 hours Result: Exhibited sub-micromolar inhibitory activities against HeLa, SiHa and MS751 with IC 50 s of 0.09 ± 0.02 μM, 0.15 ± 0.01 μM, 0.11 ± 0.03 μM. Cell Cycle Analysis Cell Line: HeLa cells [1] Concentration: 0.1, 0.2 and 0.4 μM Incubation Time: 24 hours Result: Increased cell distribution to the G2/M phase in a concentration-dependent way, arresting 24.7%, 47.6% and 71.7% of the cells in this phase at 0.1, 0.2 and 0.4 μM, respectively. Apoptosis Analysis Cell Line: HeLa cells [1] Concentration: 0.1, 0.2 and 0.4 μM Incubation Time: 24 hours Result: Induced 35.9%, 66.4% and 84.4% of cell population undergoing apoptosis at 0.1 μM, 0.2 μM, 0.4 μM, respectively. Cell Cytotoxicity Assay Cell Line: HK-2 cells [1] Concentration: 0-200 μM Incubation Time: 24 hours Result: Exhibited good renal safety profile, with IC 50 of 188 ± 16 μM in HK-2 cells.

Tubulin polymerization-IN-4 (100-1000 mg/kg; IP, single) exhibits extremely low toxicity with LD 50 over 1000 mg/kg [1]. Tubulin polymerization-IN-4 (30 and 60 mg/kg; IP; daily for 21 days) inhibits the tumor growth, with TGI of 35% and 58% at dosing 30 and 60 mg/kg [1]. Tubulin polymerization-IN-4 (30 mg/kg; IP; single) presents the modest pharmacokinetic properties [1]. Pharmacokinetic Parameters of Tubulin polymerization-IN-4 in ICR mice [1]. IP (30 mg/kg) T 1/2 (h) 1.56 ± 0.28 T max (h) 0.25 C max (μg/L) 6215 ± 308 AUC 0-t (μg/L·h) 5609 ± 347 AUC 0-∞ (μg/L·h) 5940 ± 347 V Z /F (L/kg) 11.35 ± 1.29 CL Z /F (L/h/kg) 5.05 ± 0.91 MRT (h) 1.77 ± 0.43