MPI-0479605 is an ATP competitive and selective inhibitor.

Mps1:1.8 nM

在肠癌移植模型中,每天30 mg/kg或者每4天150 mg/kg腹腔注射MPI-0479605,具有抗肿瘤活性.

在一组肿瘤细胞系中,MPI-0479605导致细胞活力显著降低,GI50范围为30至100 nM。此外,MPI-0479605通过凋亡或者有丝分裂损伤来诱发细胞生长阻滞并最终促进细胞死亡。MPI-0479605破坏SAC和染色体在有丝分裂纺锤体上的附着,进而导致染色体分离缺陷和异倍体。

In Vitro Kinase Assays and Selectivity Screening : To measure Mps1 activity, 25 ng of recombinant, full-length enzyme is incubated in reaction buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl2, 0.01% Triton X-100 and 5 μM Myelin basic protein (MBP)) containing vehicle (DMSO alone) or inhibitors. Forty μM ATP (2xKm) is added with 1 μCi [γ-33P]ATP and the reaction is incubated at room temperature for 45 minutes. Reactions are terminated with 3% phosphoric acid and transferred to P81 filter plates. Samples are washed in 1% phosphoric acid and 33P radioactivity is measured on a TopCount scintillation reader. In-house kinase assays are all carried out at 2xKm ATP concentrations. MPI-0479605 (500 nM) is also screened against a larger kinase panel.

Cells are treated for 3 or 7 days with various concentrations of MPI-0479605 and the GI50 is determined. Cell viability is measured with CellTiter-Glo. (Only for Reference)