PI3K-IN-1 (Voxtalisib Analogue) is a dual inhibitor of mTOR/PI3K, mostly for p110γ , also inhibits DNA-PK and mTOR.

PI3Kγ:9 nM, PI3Kα:39 nM, DNA-PK:150 nM, PI3Kβ:113 nM, PI3Kδ:43 nM

在GBM 39-luc细胞植入的裸鼠中,口服XL765能够抑制肿瘤细胞的生长,同时提高存活率.在BxPC-3异种移植物小鼠模型中,XL765（30 mg / kg）与氯喹（50 mg / kg）联用,能够抑制肿瘤细胞生长.

XL765对于I类PI3K有抑制作用,对于对于p110α（IC50=39 nM）,p110β（IC50=113 nM）,p110γ（IC50=9 nM）和p110δ（IC50=43 nM）。XL765通过显着抑制mTOR靶点S6,S6K和4EBP1的磷酸化,对mTOR（IC50 = 157 nM）产生抑制作用。在MIAPaCa-2细胞中,XL765处理后,产生自噬体积累。

Radioligand Binding Assays: Each tube in the A3 AR competitive binding assay contains 100 μL of membrane suspension (20 μg of protein), 50 μL of [125I]4-amino-3-iodobenzyl)adenosine-5′-N-methyluronamide (0.5 nM), and 50 μL of increasing concentrations of the test ligands in Tris-HCl buffer (50 mM, pH 7.4) containing 10 mM MgCl2 and 1 mM EDTA. Nonspecific binding is determined using 10 mM 5′-N-ethylcarboxamidoadenosine in the buffer. The mixtures are incubated at 25°C for 60 min. Binding reactions are terminated by filtration through Whatman GF/B filters under reduced pressure using a MT-24 cell harvester. Filters are washed three times with 9 mL of ice-cold buffer. Radioactivity is determined using a Beckman γ-counter, and the percent inhibition is calculated.

Cells are treated with XL765 24 hours after plating and harvested for apoptosis or autophagy assays at 24, 48, or 72 hours after XL765 treatment. Apoptosis is determined by total percentage of annexin V-positive cells by fluorescence-activated cell sorting (FACS). Acidic vesicular organelles (AVOs) are detected in XL765-treated cells by vital staining with acridine orange. The degree of AVO formation is expressed as fold increase of acridine orange fluorescence intensity (FL3) in XL765-treated cells versus control cells. (Only for Reference)