Tubulin polymerization-IN-13 (Compound 4f) is a potent inhibitor of tubulin polymerization, with an IC50 value of 0.37 μM. It displays significant anti-proliferative effects against cancer cells, inducing apoptosis and exhibiting potential antivascular activity [1].

Tubulin polymerization-IN-13 (0.005-2.8 nM) treatment inhibits tumor cell proliferation [1]. Tubulin polymerization-IN-13 (8.7-10 μM) is non-toxic in non-tumor cells [1]. Tubulin polymerization-IN-13 (1-50 nM; 24 h) treatment induces cell cycle arrest in G2/M [1]. Tubulin polymerization-IN-13 (10 nM; 24 and 48 h) treatment induces cell apoptosis [1]. Tubulin polymerization-IN-13 (10-100 nM; 24 h) treatment induces alteration of the microtubule network [1]. Cell Proliferation Assay [1] Cell Line: HeLa, HT-29, Daoy, HL-60, SEM, and Jurkat cells Concentration: 0.005-2.8 nM Incubation Time: Result: Showed IC 50 s of 2.8 nM, 2.1 nM, 0.005 nM, 2.7 nM, 0.31 nM, and 0.28 nM for HeLa, HT-29, Daoy, HL-60, SEM, and Jurkat cells, respectively. Cell Cytotoxicity Assay [1] Cell Line: Peripheral blood lymphocytes (PBL) Concentration: 8.7-10 μM Incubation Time: Result: Showed a GI 50 of 8.7 μM in quiescent lymphocytes. Cell Cycle Analysis [1] Cell Line: HeLa cells Concentration: 1, 5, 10, and 50 nM Incubation Time: 24 hours Result: Induced a G2/M arrest at 10 nM, increased G2/M cells accompanied by a strong reduction of cells in the G1 phase. Apoptosis Analysis [1] Cell Line: HeLa cells Concentration: 10 nM Incubation Time: 24 and 48 hours Result: Induced caspase-9 activation, PARP cleavage, Bcl-2 phosphorylation and Mcl-1 downregulation. Immunofluorescence [1] Cell Line: HeLa cells Concentration: 10, 50, and 100 nM Incubation Time: 24 hours Result: Showed the disorganization of microtubules at 10 nM, and much more evident at 50 and 100 nM.

Tubulin polymerization-IN-13 (intraperitoneal injection; 15 or 5 mg/kg; once every other day; 4 times) reduces tumor growth in a dose-dependent manner in an orthotopic murine tumor model [1]. Animal Model: Seven-week-old C57BL/6 female mice orthotopically injected into the mammary fat pad with E0771 mammary carcinoma cells [1] Dosage: 15 or 5 mg/kg Administration: Intraperitoneal injection; 15 or 5 mg/kg; once every other day; 4 times Result: Reduced tumor mass by 45.7% and 16.9% at 15 and 5 mg/kg, respectively. Showed no sign of toxicity and no decreasement in animal body weight.