PIM-447 dihydrochloride

PIM447 dihydrochloride (LGH447 dihydrochloride) is a potent, orally available, and selective pan- PIM kinase inhibitor, with K i values of 6, 18, and 9 pM for PIM1 , PIM2 , and PIM3 , respectively. PIM447 dihydrochloride displays dual antimyeloma and bone-protective effects. PIM447 dihydrochloride induces apoptosis

In Vitro

PIM-447 (0.05-10 µM; 24, 48 and 72 hours) has inhibitory effects in MM cells, it against sensitive cell lines with IC 50 values ranging from 0.2 to 3.3 µM (MM1S, MM1R, RPMI-8226, MM144, U266 and NCI-H929) and less sensitive cell lines with IC 50 values at 48 h >7 µM (OPM-2, RPMI-LR5, U266-Dox4 and U266-LR7). PIM-447 (0.1-10 µM; 24, 48 and 72 hours) does not induce important levels of apoptosis, when PIM447 at 5 µM, it substantially increases annexin-V levels (about 30%) in sensitive cell lines(MM1S, NCI-H929 and RPMI-8226). When PIM447 at 10 µM, it induces apoptosis in all the cell lines but to a lesser extent in OPM-2 and RPMI-LR5. PIM447 promotes the cleavage of initiator caspases, such as caspases 8 and 9, and increases the cleavage of the effector caspases 3 and 7, together with PARP cleavage in MM1S,RPMI-8226 and NCI-H929 cells. PIM447 (0.1-1 µM) increases the percentage of cells in the G0/G1 phase and decreases the proliferative phases (S and G2/M) of the cell cycle. The effects at low concentrations (0.1-1 µM) were more pronounced in MM1S cells than in OPM-2. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Cell Viability AssayCell Line: Sensitive MM cell lines: MM1S, MM1R, RPMI-8226, MM144, U266 and NCI-H929 cells Less sensitive MM cell lines: OPM-2,RPMI-LR5, U266-Dox4 and U266-LR7cells Concentration: 0.05-10 µM Incubation Time: 24, 48 and 72 hours Result: Was cytotoxic for MM cells (PIM kinases highly expressed). Apoptosis AnalysisCell Line: Sensitive MM cell lines: MM1S, NCI-H929 and RPMI-8226 cells Less sensitive MM cell lines: OPM-2 and RPMI-LR5 cells Concentration: 0.05-10 µM Incubation Time: 24, 48 and 72 hours Result: Induced cell apoptosis at higer doses, had no effects at 0.1-1 uM. Western Blot AnalysisCell Line: Sensitive MM cell lines: MM1S, NCI-H929 and RPMI-8226 cells Concentration: 0.05-10 µM Incubation Time: 24, 48 hours Result: Increased the cleavage of the effector caspases 3 and 7, and the PARP cleavage. Cell Cycle AnalysisCell Line: MM1S, OPM-2 cells Concentration: 0.1, 0.5 or 1 µM Incubation Time: 48 hours Result: Increased the cleavage of the effector caspases 3 and 7, and the PARP cleavage.

In Vivo

PIM447 (oral gavage; 100 mg/kg; 5 times/week) clearly controlls tumor progression and the serum levels of hIgλ secreted by RPMI-8226-luc cells in mouse model of bone marrow-disseminated human multiple myeloma . MCE has not independently confirmed the accuracy of these methods. They are for reference only. Animal Model: RPMI-8226-luc cells are injected intravenously into 6-week-old female NODSCID-IL-2Rγ -/- (NSG) mice Dosage: 100 mg/kg Administration: oral gavage; 100 mg/kg; 5 times/week Result: Was well tolerated, as the body weight of mice did not decrease by more than 10%. Increased bone volume density and trabecular number and reduced trabecular separation relative to vehicle group.

Form:Solid

IC50& Target:Ki: 6 pM (PIM1),18 pM (PIM1),9 pM (PIM3)