Danusertib (PHA-739358) is a small-molecule 3-aminopyrazole derivative with potential antineoplastic activity. Danusertib binds to and inhibits the Aurora kinases, which may result in cell growth arrest and apoptosis in tumor cells in which Aurora kinases are overexpressed.

RET:31 nM, FGFR1:47 nM, Abl:25 nM, TrkA:31 nM, Aurora A:13 nM

Danusertib显著抑制K562细胞的增殖,并在10天的治疗期间抑制肿瘤的生长.给予HL-60异种移植大鼠25 mg/kg Danusertib（b.d.i.v.）导致75％的肿瘤生长被抑制,并且在一只动物中完全消退.

在细胞测定中,在用Danusertib处理野生型和p53缺陷型MEF后,野生型细胞在有丝分裂（4N）中停滞,持续48小时,而p53缺陷细胞不会在4N DNA阶段停滞,持续有丝分裂,DNA变为>8N。Danusertib处理导致p53蛋白水平的上升和p21蛋白的增多,已知p53在转录水平上调节p21蛋白。Danusertib抑制其他激酶如FGFR1,Abl,Ret和Trka的活性,IC50分别为47 nM,25 nM,31 nM和31 nM。

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.

For short-term expansion assays, 1 × 103 CD34+ cells are plated in triplicates in 96-well plates containing 100 μL of serum-free medium per well supplemented with human stem-cell factor (100 ng/mL), human Flt-3 Ligand (100 ng/mL), human thrombopoietin (50 ng/mL), human interleukin-3 and -6 (IL-3 and IL-6, respectively, both 20 ng/mL), and granulocyte colony-stimulating factor (20 ng/mL) along with Danusertib at the indicated concentrations. After 5 days, an additional 100 μL of cytokine and Danusertib containing medium are added. Cell numbers within each individual well are estimated on days 3, 6, and 9 or on days 3, 6, and 12 for healthy donor samples. (Only for Reference)