AS-252424 is a potent and selective inhibitor of PI3Kγ with IC50 of 33 nM; >10 fold selectivity for PI3Kγ versus PI3Kα.

PI3Kγ:33 nM

AS-252424是一种具有选择性ATP竞争性PI3Kγ抑制剂的呋喃-2-基亚甲基噻唑烷二酮，IC50值为33 nM。对PI3Kα的抑制作用较弱，其IC50为935 nM。在对80种不同的Ser/Thr和Tyr激酶进行筛选时，除了CK2外，AS-252424在10 μM浓度下对它们均无显著的抑制作用。AS-252424以浓度依赖的方式抑制C5a介导的PKB/Akt磷酸化作用，IC50值处于亚μM或低μM范围。AS-252424还以浓度依赖的方式抑制野生型初级单核细胞和单核细胞系THP-1中MCP-1介导的趋化作用，其IC50值分别为52 μM和53 μM。[1] AS252424专门阻断胰腺癌细胞系HPAF和Capan1的增殖，通过细胞计数法评估。[2] 最近的研究表明，100 nM的AS-252424显著降低了HL-1心肌细胞中的[Ca2+]i、ICa和Ca2+瞬态。[3]

口服AS-252424，剂量为10 mg/kg时，能够使中性粒细胞的招募量中等程度减少（35%），其效果几乎与PI3Kγ缺失小鼠中观察到的结果相匹配。[1]

In vitro PI3Kγ Kinase Assay: 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 M ATP/100 nCi γ[33]ATP, final concentrations) and lipid vesicles containing 18 M PtdIns and 250 M of PtdSer (final concentrations), in the presence of AS-252424 or DMSO. Kinase reaction is stopped by adding 250 g of Neomycin-coated Scintillation Proximity Assay (SPA) beads and preceded.

After 3 hours of starvation in serum-free medium, Raw-264 macrophages are pretreated with AS-252424 or DMSO for 30 minutes and stimulated for 5 minutes with 50 nM C5a. PKB/Akt phosphorylation is monitored using phospho-Ser-473 Akt specific antibody and standard ELISA protocols.(Only for Reference)