A66 is a specific and effective p110α inhibitor(IC50=32 nM).

p110α:32 nM, p110γ:3480 nM, p110α-H1047R:43 nM, PI3KC2β:462 nM, PI4Kβ:236 nM, p110α-E545K:30 nM

In male CD1 mice, A66 (10 mg/kg) increased glucose production during the pyruvate tolerance test. In SK-OV-3 tumor tissues, A66 (100 mg/kg) effectively reduced the phosphorylation of Akt/PKB and p70 S6 kinase, inhibiting tumor cell growth.

A66 significantly inhibits the oncogenic forms of p110α, such as p110αE545K (IC50=30 nM) and p110αH1047R (IC50=43 nM), effectively reducing their activity.

IC50 values are evaluated using the PI3K (human) HTRF Assay. p85α/p110δ is obtained from Invitrogen. All other isoforms are produced in-house by co-expressing full-length human p85α with the indicated human full-length catalytic subunit containing a histidine tag at the N-terminus to allow purification. The PI3Ks are titrated and used at a concentration between their EC65-EC80 values. PI3K activity in immunoprecipitates is assayed using an antibody to the N-SH2 (N-Src homology 2) domain of p85α. Assays for other lipid kinases and protein kinases are performed by the National Centre for Protein Kinase Profiling and Invitrogen Drug Discovery Services[1].