AZD8186 is an effective and specific PI3Kβ/δ inhibitor (IC50: 4/12 nM).

PI3Kβ:4 nM, PI3Kδ:12 nM

AZD8186 强效抑制对PI3Kβ抑制敏感的MDA-MB-468细胞中的p-Akt以及对PI3Kδ抑制敏感的Jeko B细胞，其IC50分别为3 nM和4 nM。[1] 在大多数PTEN缺失的细胞系中，AZD8186 显示出优先的生长抑制活性，其GI50为<1 μM。[2]

在裸鼠体内携带PTEN缺失的PC3前列腺肿瘤异种移植物模型中，AZD8186（100 mg/kg，口服）显著抑制Akt磷酸化水平，并且显著抑制肿瘤生长。当与ABT联合使用时，AZD8186（60 mg/kg，口服）能够完全抑制肿瘤生长。[1] 在小鼠PTEN-null TNBC模型HCC70和MDA-MB-468，以及前列腺模型HID28中，AZD8186（50 mg/kg，口服）也能抑制肿瘤生长。[2] 使用AZD8186与雄激素剥夺疗法的联合治疗可导致持久的肿瘤退缩，且该效果在治疗停止后仍然持续。[3]

PI3Kα, PI3Kβ, PI3Kγ and PI3Kδ enzyme assays: The inhibition of PI3Kα, PI3Kβ, PI3Kγ and PI3Kδ human recombinant PI3K isoforms is evaluated using a Kinase-Glo Plus Assay Kit. 12 point half-log concentration-response curves with a top concentration of 100 μM are constructed by dispensing DMSO solubilised compounds into white 384-well medium-binding microplates using an Echo 555. 3 μL of the appropriate PI3K? in Tris buffer (50 mM Tris pH7.4, 0.05% CHAPS, 2.1 mM DTT, and 10 mM MgCl2) is added. The plate is covered and allowed to pre-incubate with compound for 20 minutes prior to addition of 3 μL of substrate solution containing PIP2 and ATP. The enzyme reaction is stopped after 80 minutes by the addition of Kinase Glo detection solution. Plates are covered and incubated for 30 minutes at room temperature before the luminescence signal is read using a PHERAstar plate reader. The final concentrations of DMSO, ATP and PIP2 in the assay are 2%, 8 μM, and 80 μM respectively. The final concentrations of PI3Kα, PI3Kβ, PI3Kγ and PI3Kδ are respectively 20 nM, 20 nM, 45 nM and 30 nM. For PI3Kα, PI3Kβ and PI3Kδ the concentration of active enzyme is determined as outlined in the enzyme assay tight binding limit determination section. For PI3Kγ the concentration of enzyme is determined by Bradford assay. IC50 values are calculated using Genedata Screener.

Cells are seeded in 96-well plates, at a density to allow for logarithmic growth during the 72 hour assay, and incubated overnight at 37°C, 5% CO2. Cells are treated with AZD8186 (30 to 0.003 μM) for 72 hours and cell proliferation measured by MTS. The CellTiter Aqueous Non-Radioactive Cell Proliferation Assay reagent is used in accordance with the manufacturer's protocol and Absorbance measured with Tecan Ultra instrument. Pre-dose measurements are made and the concentration needed to reduce the growth of treated cells to half that of untreated cells (GI50) values are determined.(Only for Reference)