MK-2206 2HCl

Orally administered

A431, HCC827, NCI-H292, NCI-H358, NCI-H23, NCI-H1299, Calu-6 and NCI-H460 cells

MK-2206 is currently under Phase II clinical trial for treatment of ovarian cancer, primary peritoneal cancer and fallopian tube cancer.

MK-2206 2HCl is a highly selective inhibitor of Akt1, Akt2 and Akt3 with IC50 of 8 nM, 12 nM and 65 nM, respectively.

MK-2206 is the first allosteric small molecule inhibitor of Akt to enter clinical development.

8 nM, 8 nM, 8 nM, 8 nM, 8 nM, 8 nM

MK-2206 shows 60% TGI and inhibits more than 70 % of phospho-Akt1/2 (T308 and S473) in A2780 ovarian cancer xenografts at a dose of 240 mg/kg. [1] MK-2206 exhibits significant antitumor activity in NCI-H292 xenograft in combination with erlotinib or lapatinib. [2]

[4], Akt kinases assay, Akt kinases are assayed by a GSK-derived biotinylated peptide substrate. The extent of peptide phosphorylation is determined by Homogeneous Time Resolved Fluorescence (HTRF) using a lanthanide chelate (Lance)-coupled monoclonal antibody specific for the phosphopeptide in combination with a streptavidin-linked allophycocyanin (SA-APC) fluorophore which will bind to the biotin moiety on the peptide. When the Lance and APC are in proximity, a non-radiative energy transfer takes place from the Lance to the APC, followed by emission of light from APC at 655 nm. Working Solution: 100X protease inhibitor cocktail (PIC): 1mg/mL benzamidine, 0.5 mg/mL pepstatin, 0.5 mg/mL leupeptin, 0.5 mg/mL aprotinin, 10X assay buffer: 500 mM HEPES, pH7.5, 1% PEG, 16.6 mM EDTA, 1 mM EGTA, 1% BSA, 20 mM 9-glycerol phosphate, Quench buffer 50 mM HEPES pH 7.3, 16.6 mM EDTA, 0.1% BSA, 0.1% Triton X-100, 0.17 nM labeled monoclonal antibody, 0.0067 mg/mL SA-APC, ATP/MgCl2 working solution: 1X Assay buffer, 1 mM DTT, 1X PIC, 5% glycerol, active Akt, Peptide working solution: 1X Assay buffer, 1 mM DTT, 1X PIC, 5% glycerol, 2 TM GSK biotinylated peptide. The reaction is assembled by adding 16 uL of ATP/MgCl2 working solution to the appropriate wells. MK-2206 or vehicle (1.0 uL) is added followed by 10 uL of peptide working solution. The reaction is started by adding 13 uL of the enzyme working solution and mixing. The reaction is allowed to proceed for 50 min and then stopped by the addition of 60 uL HTRF quench buffer. The stopped reactions are incubated at room temperature for at least 30 min and then read in the instrument.

480, 39

Data from [J Cell Biochem, 2010, 112, 924932], MK-2206 2HClpurchased from Selleck, Confocal microscopy images of NO formation. DAF 2 DA-loaded washed platelets (1.0109 platelets/mL) were preincubated at 378C with saline (A), and then stimulated for 1min with 0.1 (B), 1.0 (C) or 10 (D) M AEA. In Panel (E-F-G) washed platelets were preincubated with 1 M SR1 (E), 1 Mm MK2206 (F) or 20 M LY294002 (G), and then stimulated for 1min with 1.0 M AEA. In panel (H) is reported the effect of 5 mg/mL collagen, used as a positive control. All the experiments were carried out in the presence of 100 M L-arginine. NO formation was visualized by confocal microscropy as detailed in Methods.

DMSO 14 mg/mL, Water 1 mg/mL, Ethanol <1 mg/mL