Torin 1

2 years -80 in solvent

Administered via i.p. once daily

MEFs

20 mg/kg

2-10 nM [1], 2-10 nM [1], 2-10 nM [1], 2-10 nM [1], 2-10 nM [1], 2-10 nM [1]

Torin1 is efficacious at a dose of 20 mg/kg in a U87MG xenograft model and demonstrates good pharmacodynamic inhibition of downstream effectors of mTOR in tumor and peripheral tissues. [2]

mTORC1 and mTORC2 in Vitro Kinase Assays, To produce soluble mTORC1, HEK-293T cell lines that stably express N-terminally FLAG-tagged Raptor are generated using vesicular stomatitis virus G-pseudotyped MSCV retrovirus. For mTORC2, similar HeLa cells that stably express N-terminally FLAG-tagged Protor-1 are generated. Both complexes are purified by lysing cells in 50 mm HEPES, pH 7.4, 10 mm sodium pyrophosphate, 10 mm sodium beta-glycerophosphate, 100 mm NaCl, 2 mm EDTA, 0.3% CHAPS. Cells are lysed at 4 C for 30 min, and the insoluble fraction is removed by microcentrifugation at 13, 000 rpm for 10 min. Supernatants are incubated with FLAG-M2 monoclonal antibody-agarose for 1 h and then washed three times with lysis buffer and once with lysis buffer containing a final concentration of 0.5 M NaCl. Purified mTORC1 is eluted with 100 ug/mL 3xFLAG peptide in 50 mm HEPES, pH 7.4, 100 mm NaCl. Eluate can be aliquoted and stored at -80 C. Substrates S6K1 and Akt1 are purified. Kinase assays are performed for 20 min at 30 C in a final volume of 20 uL consisting of the kinase buffer (25 mm HEPES, pH 7.4, 50 mm KCl, 10 mm MgCl2, 500 um ATP) and 150 ng of inactive S6K1 or Akt1 as substrates. Reactions are stopped by the addition of 80 uL of sample buffer and boiled for 5 min. Samples were subsequently analyzed by SDS-PAGE and immunoblotting.

DMSO 2 mg/mL, Water <1 mg/mL, Ethanol <1 mg/mL

Benzo[h]-1, 6-naphthyridin-2(1H)-one, 1-[4-[4-(1-oxopropyl)-1-piperazinyl]-3-(trifluoromethyl)phenyl]-9-(3-quinolinyl)-