Danusertib (PHA-739358)

Female SCID mice

(R)-N-(5-(2-methoxy-2-phenylacetyl)-1, 4, 5, 6-tetrahydropyrrolo[3, 4-c]pyrazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide

5 uM

15 mg/kg

13 nM/79 nM/61 nM, 13 nM/79 nM/61 nM, 13 nM/79 nM/61 nM, 13 nM/79 nM/61 nM, 13 nM/79 nM/61 nM, 13 nM/79 nM/61 nM

Administration of 25 mg/kg Danusertib (b.d. i.v.) to HL-60 xenograft rats results in 75% inhibition of tumor growth with complete regression in one animal. Danusertib results in biomarker modulation accompanied by inhibition of tumor growth. This is compatible with an expected mechanism of action of aurora kinase inhibition. [1] Danusertib significantly inhibits proliferation of K562 cells and virtually suppressed tumor growth during the 10-day treatment period. [3]

Biochemical kinase Assays, The Km values for ATP and the specific substrate are initially determined, and each assay is then run at optimized ATP (2Km) and substrate (5Km) concentrations. This setting enabled direct comparison of IC50 values of Danusertib across the applied kinase selectivity screening panel for the evaluation of the selectivity profile.

474, 55

, , Dr. Yong-Weon Yi from Georgetown University Medical Center, Danusertib (PHA-739358)purchased from Selleck, For MTT assays, cells (2, 000 ca. 5, 000 cells/well) were subcultured into 96-well plates according to their growth properties. Cell proliferation was assayed at 72 hr after treatment of PHA-739358 by adding 20 l of 5 mg/ml 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) solution per 100 l of growth medium. After incubating for 3-4 h at 37C, the media were removed and 150 l/well of MTT solvent (either absolute DMSO or isopropanol containing 4 mM HCl and 0.1% Nonidet-40) was added to dissolve the formazan. The absorbance of each well was measured by ELx808 (BioTek, Winooski, VT) or Wallac Victor2 (Perkin-Elmer Life Sciences, Boston, MA) Microplate Reader. Viable cells are presented as percent of control, vehicle-treated cells.

DMSO 95 mg/mL, Water <1 mg/mL, Ethanol <1 mg/mL