Description

CAY10505 is a PI3Kγ inhibitor with IC50 of 30 nM.

Targets

IC₅₀

In Vitro

CAY10505 selectively inhibits PI3Kγ isoform, with an IC50 of 30 nM better than the PI3Kα, β, and δ isoforms, with IC50 of 0.94, 20 and 20 μM, respectively. Tested against a panel of 80 other kinases including casein kinase 2, CAY10505 significantly inhibits only the unrelated casein kinase 2 (CK20) with an IC50 of 20 nM. CAY10505 also inhibits the phosphorylation of the PI3K substrate PKB/Akt in mouse macrophages with an IC50 of 228 nM. CAY10505 inhibits C5a-mediated?PKB/Akt phosphorylation?in Raw-264?macrophages, with an IC50 of 0.23?nM. In human monocytic cell line THP-1, MCP-1, binding to the GPCR chemokine receptor CCR2, strongly induces phosphorylation of PKB/Akt, which is effectively inhibited by 26 at IC50 values as low as 0.4 μM. CAY10505 inhibits MCP-1-mediated chemotaxis in wild-type primary monocytes in a concentration-dependent manner with an IC50 of 52 μM, as well as in the monocytic cell line THP-1 with an IC50 of 53 μM. ^[1]

In Vivo

Oral administration of CAY10505 at 10 mg/kg results in moderate reduction of neutrophil recruitment by 35%, almost matching the result observed in PI3Kγ-deficient mice. ^[1] Five weeks of deoxycorticosterone acetate salt (DOCA) administration are followed by 7 days of daily administration of PI3Kγ inhibitor CAY10505 at a dose of 0.6 mg/kg (p.o.), which significantly improves acetylcholine-induced endothelium dependent relaxation, serum nitrate and (or) nitrite level, glutathione level, and the vascular endothelial lining in hypertensive rats. ^[2]

Clinical Trials

Features

PI3Kγ lipid kinase assay

A PI3Kγ lipid kinase assay, based on the neomycin-coated scintillation proximity assay (SPA) bead technology, is performed in 384-well plates using ATP/[γ³³P]ATP and PtdIns as substrates. Kinase assays for IC50 value determinations of CAY10505 with PI3Kα, PI3Kβ, and PI3Kδ are carried out.

Cell Lines

Bone marrow cells

Concentrations

~10 mM

Incubation Time

3 hours

Methods

Bone marrow cells are isolated, prepared, and stimulated. For in vitro chemotaxis, 10⁷ 5-day-derived BMDMs are suspended in 1 mL of medium containing 0.5% BSA and CAY10505 or DMSO and applied to the upper chamber of a transwell, 5 μm pore size, chemotaxis plate. An amount of 600 μL of medium containing MCP-1/CCL2 and inhibitors or DMSO is added to the lower chamber. After 3 hours of incubation at 37 °C and 5% CO₂, the number of cells in the lower chamber is quantified with a Coulter.

Animal Models

Wistar albino rats

Formulation

Doses

0.6 mg/kg

Administration

Administered via p.o.