Torin 2(IC50=0.25 nM), a specific and effective mTOR inhibitor, is the 800-fold greater specific activity for mTOR than PI3K and improves pharmacokinetic properties. The EC50 of Torin 2 for ATM/ATR/DNA-PK inhibition is 28 nM/35 nM/118 nM, respectively.

mTOR:0.25 nM, ATR:35 nM(EC50), DNA-PK:118 nM(EC50), ATM:28 nM(EC50)

在Th-MYCN 小鼠体内,Torin 2（20 mg/kg）消除MYCN肿瘤,减少MYCN蛋白质水平,并诱导细胞凋亡.在小鼠肝微粒体稳定性试验中,Torin 2表现出>95%的药效学响应,半衰期为11.7分钟.在雄性Swiss albino小鼠中,静脉内或口服Torin 2后,体内表现出最好的生物利用度为51%,半衰期为0.72小时,和低清除率是19.6 mL/min/kg.

在MZ-CRC-1和TT细胞中, Torin 2（< 50 nM）引起细胞的的活性明显减少,Torin 2（100 nM）使细胞的迁移明显减少。Torin 2抑制mTORC1,因此通过促进其核转运激活TFEB,EC50为1.666 mM。Torin 2与PI3Kγ具有相同的结合模式,V882作为铰链结合点,在内部疏水袋Y867,D841 和D964再提供三个作用于氨基比林侧链的氢键,类似于mTOR的Y2225,D2195 和D2357。

mTOR and PI3K Cellular Assays: Cellular IC50 values for mTOR are determined using p53?/? MEFs. Cells are treated with vehicle or increasing concentrations of Torin 2 for 1 h and then lyse. Phosphorylation of S6K1 Thr-389 is monitored by immunoblotting using a phospho-specific antibody. Meanwhile, cellular IC50 values for PI3Ka are determined based on phosphorylation of Akt Thr-308 in p53?/?/mLST8?/? MEFs or human PC3 cells expressing the S473D mutant of Akt1.

For viability, MZ-CRC-1 and TT cells are seeded in quadruplicate in 96-well plates (1.0×104 cells per well) in culture media with 2.5% and 4% FBS, respectively. After 24 hours, cells are treated with Torin 2. At the indicated time point, cells are incubated for 3 hours with 10 μL of CellTiter96 AQueous One solution in 100 μL of culture media and absorbance is measured at 490 nm.(Only for Reference)