SGI-1776 free base

2 years -80 in solvent

Administered via oral gavage (PO) on a daily x 5/week or twice/week schedule

MV-4-11, MOLM-13, and OCI-AML-3 cell lines

0.1-10 uM

5% dextrose

In addition to Pim, SGI-1776 also potently targets FLT3 (IC50 = 44nM). Treatment of AML cells with SGI-1776 results in a concentration-dependent induction of apoptosis. Importantly, SGI-1776 is also cytotoxic in AML primary cells, irrespective of FLT3 mutation status and results in Mcl-1 protein decline. [1]Treatment of CLL cells with SGI-1776 results in a concentration-dependent induction of apoptosis. SGI-1776 induces apoptosis in CLL and that the mechanism involves Mcl-1 reduction. Apoptosis induction coupled with the inhibition of RNA synthesis is observed in CLL cells treated with SGI-1776. [2] SGI-1776 exhibites cytotoxic activity in vitro with a median relative IC50 of 3.1 mM. SGI-1776 induces tumor growth inhibition meeting criteria for intermediate EFS T/C activity in 1 of 39 evaluable models. In contrast, SGI-1776 induces complete responses of subcutaneous MV4, 11. [3]

24 hours

Cells are cultured in IMDM (ATCC) supplemented with 10% FBS and grown in a 37C incubator with 5% CO2. Cells are routinely tested for Mycoplasma infection using a commercially available kit. Cells are treated with DMSO or various concentrations of SGI-1776 for 24 hours. Cells (1x106) are washed, then resuspended in 100 uL of annexin binding buffer, mixed with 5 uL of FITC solution and 5 uL of propidium iodide (PI, 50 ug/mL) solution. For each sample, 1x104 cells are measured using a Becton Dickinson FACSCalibur flow cytometer.

SGI-1776 free base is a novel ATP competitive inhibitor of Pim1 with IC50 of 7 nM in a cell-free assay, 50- and 10-fold selective versus Pim2 and Pim3, also potent to Flt3 and haspin. SGI-1776 induces apoptosis and autophagy.