GNE-317, a PI3K/mTOR inhibitor, can pass through the blood-brain barrier (BBB).

GNE-317，一种通过GDC-0980合成的环氧乙烷衍生物，旨在降低底物对外排转运蛋白的亲和力。体外实验中，GDC-0980与GNE-317在使用GL261细胞系的MTS细胞毒性实验中展现出了相似的特性。

小鼠通过i.c.接种含有GL261-GFP-Luc细胞后七天，每日一次接受最大耐受剂量的GDC-0980（7.5 mg/kg）、GNE-317（30 mg/kg）或安慰剂治疗。通过每周一次的生物发光成像追踪GL261肿瘤生长情况。GDC-0980、GNE-317或安慰剂三组治疗中，GL261肿瘤生长无显著差异。数据显示，这些化合物在诱导GL261细胞系细胞死亡方面效果有限。尽管GNE-317在化合物传递和治疗靶向效能上更为突出，但在GL261肿瘤治疗中并不有效。

Cellular viability assays are set up in a 96-well format with 2000 GL261-GFP-Luc cells plated per well in the culture conditions. GL261, an aggressive C57BL/6J-derived glioma line, is transfected with both green fluorescent protein (GFP) and luciferase (Luc) from separate plasmids.GL261-GFP-Luc cells are cultured in Dulbecco's modified Eagle's medium supplemented with 10% FBS and Penicillin/Streptomycin (100 U/mL) at 5% oxygen, and are selected by 4 mg/mL Puromycin and 4 mg/mL G418. Suspend GNE-317 in DMSO and then diluted with the medium.GL261-GFP-Luc cells are incubated in the presence of drug or vehicle for 48 hours, and viability was assessed by MTS assay. Results were detected using a Synergy Mx automated plate reader,which are set up absorbance at 490 nm and used to determine viability and at 650 nm to account for the background. Numerical values from drug-treated wells are normalized to the values of vehicle-treated wells to yield percent survival.

7-week-old C57BL/6J mice are implanted GL261-GFP-Luc cells. When tumors reach 5e7 photons/s/cm2/sr (radiance), mice are orally administered the maximum tolerated the dose,which is defined as <10% drop in mice bodyweight dose, GDC-0980 for 7.5 mg/kg, GNE-317 for 30 mg/kg or vehicle once daily for 3 days. At 1 or 6 hours after the third dose, mice are euthanized with carbon dioxide and perfused with 30 mL PBS.