Alpelisib (BYL-719) is a PI3Kα inhibitor (IC50=5 nM) with selective, potent, and oral activity. Alpelisib inhibits PI3Kβ/γ/δ with low activity (IC50=250/290/1200 nM). Alpelisib has antitumor activity and is targeted to PIK3CA mutant tumors.

p110α:5 nM (cell free), p110γ:250 nM (cell free)

方法：骨肉瘤细胞系 MG-63、HOS、MOS-J 和 POS-1 用 Alpelisib (0-50 µM) 处理 72 h，使用 XTT assay 检测细胞活力。
结果：Alpelisib 以剂量依赖性方式显著抑制所有骨肉瘤细胞系的细胞生长，IC50 范围为 6-15 µM，IC90 范围为 24-42 µM。[1]
方法：PIK3CA 野生型细胞 (SNU638 和 SNU668) 和三个 PIK3CA 突变体细胞 (SNU601、AGS 和 MKN1) 用 Alpelisib (5 µM) 处理 24 h，使用 Flow cytometry 检测细胞周期情况。
结果：无论 PIK3CA 突变状态如何，Alpelisib 处理都诱导 G0/G1 细胞周期停滞。在 PIK3CA 突变细胞 (AGS 和 MKN1) 中，亚 G1 组分显著增加，表明 Alpelisib 在这些细胞系中增加了细胞凋亡。[2]

方法：为检测体内抗肿瘤活性，将 Alpelisib (12.5-50 mg/kg，methylcellulose 0.5%) 口服给药给携带人骨肉瘤 HOS-MNNG 的 Rj:NMRI-nude 小鼠，每天一次，持续二十二天。
结果：Alpelisib 以剂量依赖性方式显著减少肿瘤体积。[1]
方法：为研究对胶原蛋白的调节作用，将 Alpelisib (12.5-50 mg/kg) 口服给药给携带 Rat1-myr-p110α 肿瘤皮下异种移植的 nude 小鼠，每天一次，持续八天。
结果：12.5、25、50 mg/kg 的 Alpelisib 治疗具有良好的耐受性，并产生了剂量依赖性和统计学上显著的抗肿瘤效果，T/C 为 14.1%，消退为 9.6% 和 65.2%。[3]

To evaluate the isoform-specific potency of NVP-BYL719 in a cell-based system, an N-terminally myristoylated form of each PI3K class IA isoform was expressed in Rat1 fibroblasts. The retroviral expression plasmid pBabePuro containing human p110α, p110β, and p110δ with an N-terminal myristoylation (myr) signal followed by an HA-tag were generated. Successfully infected Rat1 cells were selected in medium containing 4 μg/mL of puromycin, expanded and characterized for expression of the p110 isoforms. Transgenic expression of the myristoylated protein was confirmed by increased levels of phosphorylated Akt [1].

All in life experimentation and efficacy studies were conducted as described previously. Tumor xenografts were grown subcutaneously or orthotopically in nude mice or nude Rowett rats (Hsd: RH-Fox1rnu) by injection of 3 × 10^6 to 1 × 10^7 cells or implantation of tumor fragments of approximately 50 mg. Tumor-bearing animals mice were treated with either vehicle control, NVP-BYL719, or NVP-BKM120 (p.o., every day) at the doses indicated [1].