Hence, we are able to examine the effects of KLB perturbation in an aggressive

A determination of the crystal structure of the T315I Abl kinase domain in complex with PHA 739358 showed that the drug interacts with the active conformation オーダー ABT-737 of Abl kinase. Currently, preliminary evidence for anti tumor activity of PHA 739358 has been seen in various advanced refractory can cers, and phase II studies in solid tumors are ongoing. In this report, we performed preclinical studies in the presence of stroma in vitro as well as in vivo, to explore the application of PHA 739358 for treatment of a variety of primary human acute lymphoblastic leukemia cells including those belonging to the Ph positive ALL sub class and harboring the T315I mutation. We conclude that PHA 739358 could be considered for the treatment of patients with different subtypes of ALL in combin ation with other drugs to potentiate its cytostatic and cytotoxic effects.<br><br> Results PHA 739358 reduces viability of acute lymphoblastic leukemia cells including those with the BcrAbl T315I mutation To determine AEB071 1058706-35-6 the impact of the BcrAbl status on the effi cacy of PHA 739358, we treated human ALL cells includ ing BLQ1, Pt2, UCSF02, TXL2, US7, US7R and mouse 8093 and Bin2 cells with increasing concentrations of PHA 739358 for 72 hours. In Phase I II clinical trials, a Cmax of 4 6 uMh was observed for CML patients harboring the T315I mutation when PHA 739358 was administered at 330 mgm2day. Therefore, we used clinically relevant and achievable concentrations of up to 5 uM PHA 739358 in our experiments. As shown in Figure 1, increasing concentrations of PHA 739358 caused a cytotoxic effect on all the leukemia cells tested as measured by the decreased viability of the cultures.<br><br> There buy AG-014699 was no correlation between the type of ALL and sensitivity to the drug. Compared to human leukemia cells, mouse 8093 and Bin2 cells were signifi cantly more sensitive to PHA 739358. Although these murine BcrAbl ALL cells contain an identical transgene, they also exhibited different sensitivity to this drug. PHA 739358 induces apoptosis and leads to an accumulation of cells with 4N DNA content The ability of PHA 739358 to induce apoptosis was mea sured by Annexin VPI staining in Pt2 and UCSF02 cells treated with increasing concentrations of the drug for 48 hours. As demonstrated in Figure 2A, PHA 739358 induced apoptosis both in Pt2 and UCSF02 cells.<br><br> Since in hibition of Aurora kinases causes endoreduplication and polyploidy, we assessed DNA content at different time points in Ph positive BLQ1 and Ph negative US6 cells trea ted with PHA 739358. Mutations and deletions of p53 are rare in ALL and of the samples examined here, only US6 had defective p53 function. In agreement with previous findings using Aurora kinase inhi bitors in other types of cancer cells, PHA 739358 caused accumulation of BLQ1 and US6 cells with more than or equal to 4 N DNA content as early as 16 hours. Moreover, 1 uM PHA 739358 generated polyploid cells and produced a significant reduction in viability, as assessed by the percentage of cells in the sub G1 DNA content. PHA 739358 targets both BcrAbl and Aurora kinase activities PHA 739358 was reported to inhibit both BcrAbl kinase and Aurora kinase in vitro, whereas dasatinib targets BcrAbl and Src family kinases.