ML141 (CID-2950007) is an effective, specific and reversible non-competitive inhibitor of Rho family GTPase cdc42 (IC50: 200 nM).

CDC42:200 nM

In NOD/SCID mice carrying MDA-MB 231-derived tumors, ML141 (1 mg/day, intraperitoneally) inhibits the growth of these tumors by suppressing Cdc42, thereby enhancing the effectiveness of TMX. Additionally, ML141 (10 mg/kg, intraperitoneally) increases the mobilization of hematopoietic stem and progenitor cells induced by granulocyte colony-stimulating factors.

ML141 significantly protects against apoptosis damage induced by metformin in neuroblastoma. It enhances the ability of caffeine to inhibit cell growth through the induction/suppression of cell death/division. Additionally, ML141 dose-dependently reduces the invasion of Klebsiella pneumoniae.

Equilibrium binding assay : Wild-type GST-Cdc42 (4 μM) is bound to GSH-beads overnight at 4°C. Cdc42 on GSH-beads is depleted of nucleotide by incubating with 10 mM EDTA containing buffer for 20 min at 30°C, washing twice with NP- HPS buffer, then re-suspended in the same buffer containing 1 mM EDTA/or 1 mM MgCl2, 1 mM DTT and 0.1% BSA. Cdc42 unbound sites are blocked by incubation of protein–bead complex for 15 min at RT. Thirty μL of this suspension is incubated with 20 mM inhibitor for 3 min at RT and added 30 μL of various concentrations of ice cold BODIPY-FL-GTP. Samples incubated at 4° C for 45 min and binding of fluorescent nucleotide to enzyme is measured using an Accuri flow cytometer. Raw data are exported and plotted using GraphPad Prism software.

Cells are incubated with 500 nM Calcein-AM and 1 µM PI for 15 min, after which live cells and dead cells (represented by positivity of Calcein-AM and PI staining, respectively) are counted utilizing the adherent cell Celigo™ cytometer. (Only for Reference)