AS-604850 is a specific and ATP-competitive PI3Kγ inhibitor (IC50: 250 nM), 18-fold more selective for PI3Kγ than PI3Kα, and over 80-fold selectivity for PI3Kγ than PI3Kδ/β.

PI3Kγ:250 nM

AS-604850是一种与ATP竞争的PI3Kγ抑制剂，其Ki值为0.18μM。该化合物是PI3Kγ的同型选择性抑制剂，对PI3Kδ和β具有超过30倍的选择性，对PI3Kα则具有18倍的选择性。（PI3Kα: IC50 = 4.5μM, PI3Kγ和β: IC50 > 20μM）AS-604850能够抑制RAW264小鼠巨噬细胞中C5a介导的PKB磷酸化，其IC50为10μM。此外，AS-604850以浓度依赖性方式阻断Pik3cg+/+单核细胞中MCP-1介导的趋化作用，IC50为21mM，但不影响Pik3cg-/-细胞的趋化作用，表明AS-604850通过PI3Kγ发挥作用。[1] AS-604850减少了大鼠肝细胞中甘胆酸诱导的Akt磷酸化和凋亡。此外，AS-604850减轻了HepG2 Ntcp和Huh7-Ntcp细胞中胆盐诱导的凋亡。[2] AS604850能够浓度依赖地抑制EoL-1细胞和血嗜酸性粒细胞对血小板激活因子（PAF）的趋化反应。[3]

AS-604850通过口服给药，在ED50为42.4 mg/kg的条件下，能有效降低RANTES引起的腹膜中性粒细胞的招募。在硫代乙酸盐引起的腹膜炎模型中，10 mg/kg剂量的AS-604850能使中性粒细胞招募减少31%。[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.

Hepatocyte cultures are treated with diluent (DMSO), 25 μM TLC, 250 μM TCDC, 50 μM GCDC, or 50 ng/ml Fas for 2-4 hours HepG2-Ntcp and Huh7-Ntcp cells are treated with DMSO, 20 μM TLC, 75 μM TCDC or GCDC, 200 μM etoposide or 200 ng/ml TNFa plus 28 ng/ml actinomycin D for 2-4 hours. The number of apoptotic cells is determined morphologically using fluorescent staining and expressed as % of cells. Apoptosis is confirmed in human cell lines by determination of caspase-3/-7 activity and in rat hepatocytes by detection of the 17 kDa proteolytic cleavage fragment of caspase-3 by immunoblotting; equal protein loading is monitored by immunoblotting for actin.(Only for Reference)