Afuresertib (GSK2110183) is an orally bioavailable inhibitor of the serine/threonine protein kinase Akt (protein kinase B) with potential antineoplastic activity.

Akt3:2.6 nM(Ki), Akt1:0.08 nM(Ki), Akt2:2 nM(Ki)

用10,30和100 mg/kg Afuresertib处理负荷SKOV3卵巢肿瘤异种移植物的小鼠,分别导致凝血酶形成指数为23%,37%和97%.每天给负荷BT474乳腺肿瘤异种移植物的小鼠,口服10,30或100 mg/kg Afuresertib,分别导致凝血酶形成指数为8%,37%和61%.

总体上65%的血液学细胞系对Afuresertib敏感,EC50 < 1 μM。在测试的实体肿瘤细胞系中,21%对Afuresertib的EC50 <1 μM。Afuresertib抑制E17K AKT1突变蛋白的激酶活性,EC50为0.2 nM。Afuresertib显示对多种AKT底物磷酸化水平（包括GSK3b,PRAS40,FOXO和胱天蛋白酶9）的浓度依赖性效应。

Potency (Ki*) of afuresertib: The true potency (Ki*) of the inhibitor is initially determined at low enzyme concentrations (0.1 nM AKT1, 0.7 nM AKT2, and 0.2 nM AKT3) using a filter binding assay and then confirmed with progress curve analysis. In the filter binding assay, a pre-mix of enzyme plus inhibitor is incubated for 1 h and then added to a GSKα peptide (Ac-KKGGRARTSS-FAEPG-amide) and [γ33P] ATP. Reactions are terminated after 2 h and the radio labeled AKT peptide product is captured in a phospho-cellulose filter plate. Progress curve analysis utilizes continuous real-time fluorescence detection of product formation using the Sox-AKT-tide substrate (Ac-ARKRERAYSF-d-Pro-Sox-Gly-NH2).

A 3-day proliferation assay using CellTiter-Glo is performed to measure the growth inhibition by the compounds at 0-30 μM. Cell growth is determined relative to untreated (DMSO) controls. EC50's are calculated from inhibition curves using a 4- or 6-parameter fitting algorithm in the Assay Client application.(Only for Reference)