AS-605240 is an effective and specific inhibitor of PI3Kγ (IC50/Ki: 87.8 nM).

PI3Kγ:87.8 nM

在肥胖症诱导的糖尿病ob/ob鼠模型中,AS-605240（10 mg/kg）会降低血糖水平, 使胰岛素敏感性和葡萄糖耐量显著提高,且对体重无影响,30 mg/kg剂量的效果更好,对体重只有轻微影响.AS-605240使ATMs量和MCP-1循环水平降低. 在αCII诱导的关节炎中,AS-605240（50 mg/kg）保护其免受 αCII-IA症状.在胶原诱导的关节炎鼠模型中,AS-605240（50 mg/kg）也抑制关节炎症和损伤.AS-605240可使RANTES诱导的患腹膜炎鼠体内的中性粒细胞趋化性降低（ED50：9.1 mg/kg）.

在骨髓单核细胞中,AS-605240（1 μM）对MCP-1或CSF-1诱导的PKB磷酸化有抑制作用。在鼠的SC-CA1突触中,AS-605240（100 nM）消除NMDAR LTD,但对mGluR LTD和LTP无影响。AS-605240对PI3Kα/β/δ（IC50：60/270/300 nM）也有抑制作用。AS-605240还抑制C5a调节的PKB磷酸化（IC50：90 nM）。

In vitro PI3K lipid kinase assay: (1) For PI3Kγ: human PI3Kγ (100 ng) is incubated at RT with kinase buffer (10 mM MgCl2, 1 mM β-glycerophosphate, 1 mM DTT, 0.1 mM Na3VO4, 0.1% Na Cholate and 15 μM ATP/100 nCi γ[33P]ATP, final concentrations) and lipid vesicles containing 18 μM PtdIns and 250 μM of PtdSer (final concentrations), in the presence of AS-605240 or DMSO. Kinase reaction is stopped by adding 250 μg of Neomycin-coated Scintillation Proximity Assay (SPA) beads. (2) For PI3Kα, β, and δ: varying amounts of ATP are incubated with the different purified PI3K isoforms and saturating concentrations of PtdIns. Consequently, IC50 determinations with PI3Kα, β, and δ, to evaluate inhibitor selectivity are performed as follows: 60 ng of PI3Kα are incubated at RT with kinase buffer, as described for PI3Kγ (but containing 89 μM ATP/300 nCi γ[33P]ATP and no Na Cholate, instead) and lipid vesicles containing 212 μM PtdIns and 58 μM of PtdSer. 100 ng of PI3Kβ are incubated at RT with kinase buffer (containing 70 μM ATP/300 nCi γ[33P]ATP, 4 mM MgCl2 and no Na Cholate) and lipid vesicles containing 225 μM PtdIns and 45 μM of PtdSer. 90 ng of PI3Kδ are incubated with kinase buffer (containing 65 μM ATP/300 nCi γ[33P]ATP, 1 mM MgCl2, and no Na Cholate) and lipid vesicles containing 100 μM PtdIns and 170 μM of PtdSer. The reactions are stopped after 2 hours.

After a 3-hour starvation in serum-free medium, Cells are pretreated with AS-605240 or DMSO for 30 min and stimulated for 5 min with 50 nM of C5a. PKB phosphorylation is monitored using phosphorylated Ser473 Akt-specific antibody and standard ELISA protocols. (Only for Reference)