Cell lines

MOLM-13 cells

Preparation method

The solubility of this compound in DMSO is >16.6mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

0.01-0.3 μM; 24, 48, and 72 h; 37℃

Applications

In MOLM-13 cells, KW-2449 inhibited the phosphorylation of FLT3 (P-FLT3) and its downstream molecule phospho-STAT5 (P-STAT5) in a dose-dependent way. Furthermore, KW-2449 increased the percentage of cells in the G1 phase and reduced the percentage of cells in the S phase, resulting in the increase of apoptotic cell population.

Animal models

SCID mice bearing the subcutaneous MOLM-13 tumor

Dosage form

2.5, 5.0, 10, and 20 mg/kg; orally administered; twice a day for 14 days

Application

In SCID mice bearing the subcutaneous MOLM-13 tumor, KW-2449 completely reduced the levels of P-FLT3 and P-STAT5 in the tumor from 4 to 12 hours. While the phosphorylation of FLT3 and STAT5 returned to almost the basal level at 24 hours. KW-2449 showed a potent and significant antitumor effect in a dose-dependent way.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

KW 2449 is a multikinase inhibitor of FLT3, ABL, ABL-T315I, and Aurora kinase with IC50 values of 6.6nM, 14nM, 4nM and 48nM, respectively [1].

KW-2449 is a potent kinase inhibitor against FLT3, ABL, T315I-mutant ABL (ABL-T315I) tyrosine kinases as well as Aurora kinase. In vitro assay shows KW-2449 has the potency of growth inhibition with different mechanisms. In leukemia cells with FLT3 mutations, KW-2449 down-regulates phosphorylated-FLT3/STAT5, induces G1 arrest and apoptosis. KW-2449 also inhibits tumor growth in FLT3-mutated xenograft model. While in leukemia cells with wild-type FLT3, KW-2449 reduces phosphorylated histone H3, induces G2/M arrest and apoptosis. However, it is reported that the AML patients treated with LT3 inhibitors, such as KW-2449, have only partial FLT3 inhibition in vivo. And the plasma level of FL (the ligand of FLT3) appears to rise after chemotherapy, which impeding the efficacy of the FLT3 inhibitors [1, 2].

References:
[1] Shiotsu Y, Kiyoi H, Ishikawa Y, Tanizaki R, Shimizu M, Umehara H, Ishii K, Mori Y, Ozeki K, Minami Y, Abe A, Maeda H, Akiyama T, Kanda Y, Sato Y, Akinaga S, Naoe T. KW-2449, a novel multikinase inhibitor, suppresses the growth of leukemia cells with FLT3 mutations or T315I-mutated BCR/ABL translocation. Blood. 2009 Aug 20;114(8):1607-17.
[2] Sato T, Yang X, Knapper S, White P, Smith BD, Galkin S, Small D, Burnett A, Levis M. FLT3 ligand impedes the efficacy of FLT3 inhibitors in vitro and in vivo. Blood. 2011 Mar 24;117(12):3286-93.