IC-87114 is a specific PI3Kδ inhibitor(IC50=0.5 μM).

PI3Kγ:29 μM, PI3Kδ:0.5 μM

In a mouse model of inflammation, IC87114 inhibits the extracellular secretion of proteases prompted by TNF1α stimulation.

In human neutrophils, IC87114 at a concentration of 5 μM effectively inhibits the biosynthesis of phosphatidylinositol 3-phosphate (PIP3) and chemotaxis stimulated by N-formylmethionyl-leucyl-phenylalanine (fMLP). Furthermore, in primary cells from acute myeloid leukemia (AML) patients, such as bone marrow mononuclear cells (BMMCs), IC87114 at 10 μM concentration suppresses constitutive and Flt-3-induced Akt phosphorylation as well as cell proliferation. In addition, IC87114 inhibits proliferation and interferon-gamma (IFN-γ) production in mouse CD62L positive (naive) and CD62L negative (effector/memory) CD4+ T cells stimulated by anti-CD3.

PI3K kinase assay: Phosphatidylinositol-(4,5)-bisphosphate (PIP2) containing phospholipid liposomes are prepared. Briefly, bovine PIP2 and phosphatidylserine (1:2 molar ratio) are vacuum-dried and resuspended at 1 mM PIP2 in 20 mM HEPES-KOH, pH 7.4, 50 mM NaCl, and 5 mM EDTA. The lipid suspension is subjected to a brief sonication, followed by 5 freeze-thaw cycles and then 20 extrusion cycles to produce the liposomes. The assay is conducted in 60 μL reaction volumes in 20 mM HEPES, pH 7.4, buffer containing 1 nM PI3K, 1 μM PIP2, 200 μM ATP, 1 μCi [γ-32P]ATP, 5 mM MgCl2, plus 50 μg/mL horse IgG as carrier protein. The reaction is incubated for 10 min at room temperature, quenched in 140 of 1 M K2PO4, 30 mM EDTA, pH 8.0, captured onto a 96-well polyvinylidene difluoride filter plate, and washed five times with 1 M K2PO4. The filter is allowed to dry completely, and the bound radioactivity is quantitated. IC87114 dilutions are assayed in a final concentration of 1% (w/w) DMSO.

For AML cell proliferation assay, BMMCs are isolated and cultured in α-medium with 5% fetal calf serum (FCS) with or without FLT-3 ligand (10 ng/mL) for 48 hours and with or without IC87114. [3H]-thymidine (1 μCi [37 kBq]) is added for a final 6 hours and the amount of radioactivity incorporated is determined by trichloracetic acid precipitation. CD34+ cells from cord blood are cultured in stem cell factor (SCF; 20 ng/mL), FLT-3 ligand (10 ng/mL), and Tpo (20 nM) for 48 hours with or without 10 μM IC87114 and pulsed for 12 hours with [3H]-thymidine. (Only for Reference)