Torin 1

Catalog No.S2827

Torin 1 is a potent inhibitor of mTORC1/2 with IC50 of 2 nM/10 nM in cell-free assays; exhibits 1000-fold selectivity for mTOR than PI3K.

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Torin 1 Chemical Structure

Torin 1 Chemical Structure
Molecular Weight: 607.62

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Related Compound Libraries

mTOR Inhibitors with Unique Features

Product Information

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  • Research Area
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description Torin 1 is a potent inhibitor of mTORC1/2 with IC50 of 2 nM/10 nM in cell-free assays; exhibits 1000-fold selectivity for mTOR than PI3K.
Targets mTORC1 [1]
(Cell-free assay)
mTOR [2]
(Cell-free assay)
DNA-PK [2]
(Cell-free assay)
mTORC2 [1]
(Cell-free assay)

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IC50 2 nM 4.32 nM 6.34 nM 10 nM
In vitro Torin1 inhibits phosphorylation of mTORC1 and mTORC2 substrates in cells at concentrations of 2 and 10 nM, respectively. Moreover, Torin1 exhibits 1000-fold selectivity for mTOR over PI3K (EC50 = 1800 nM) and exhibits 100-fold binding selectivity relative to 450 other protein kinases. [1] [2] Torin1 causes cell cycle arrest through a rapamycin-resistant mechanism that is also independent of mTORC2. Torin1 disrupts mTORC1-dependent phenotypes more completely than rapamycin. Rapamycin-resistant functions of mTORC1 are required for cap-dependent translation. [1] In a recent study, it is reported Torin1 increases neurotensin secretion and gene expression through activation of the MEK/ERK/c-Jun pathway in the human endocrine cell line BON. [3]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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... Click to View More Cell Line Experimental Data

In vivo 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]
Features

Protocol(Only for Reference)

Kinase Assay: [1]

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 β-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 μg/mL 3×FLAG 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 μL consisting of the kinase buffer (25 mm HEPES, pH 7.4, 50 mm KCl, 10 mm MgCl2, 500 μm ATP) and 150 ng of inactive S6K1 or Akt1 as substrates. Reactions are stopped by the addition of 80 μL of sample buffer and boiled for 5 min. Samples are subsequently analyzed by SDS-PAGE and immunoblotting.

Cell Assay: [1]

Cell lines MEFs
Concentrations ~250 nM
Incubation Time 4 days
Method Cell viability is assessed with the CellTiter-Glo Luminescent Cell Viability Assay. On Day 0, 96-well plates are seeded with 500 cells per well and grown overnight. On Day 1, cells are treated with the appropriate compounds and subsequently analyzed on Days 3-5. For analysis, plates are incubated for 60 min at room temperature; 50 μL of CellTiter-Glo reagent is added to each well, and plates are mixed on an orbital shaker for 12 min. Luminescence is quantified on a standard plate luminometer.

Animal Study: [2]

Animal Models U87MG xenograft model
Formulation Torin1 powder is first dissolved at 25 mg/mL in 100% N-methyl-2-pyrrolidone and subsequently diluted 1:4 with sterile 50% PEG400 to a final concentration of 5 mg/mL
Dosages 20 mg/kg
Administration Administered via i.p. once daily

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Thoreen CC, et al, J Biol Chem, 2009, 284(12), 8023-8032.

[2] Liu Q, et al, J Med Chem, 2010, 53(19), 7146-7155.

view more

Chemical Information

Download Torin 1 SDF
Molecular Weight (MW) 607.62
Formula

C35H28F3N5O2

CAS No. 1222998-36-8
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 2 mg/mL (3.29 mM)
Water <1 mg/mL (<1 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 30% PEG 400+0.5% Tween80+5% Propylene glycol 30mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name Benzo[h]-1,6-naphthyridin-2(1H)-one, 1-[4-[4-(1-oxopropyl)-1-piperazinyl]-3-(trifluoromethyl)phenyl]-9-(3-quinolinyl)-

Customer Product Validation(4)


Click to enlarge
Rating
Source Mol Cell Biol 2014 34(24), 4474-84. Torin 1 purchased from Selleck
Method Immunoprecipitation
Cell Lines HeLa cells
Concentrations 1 mM
Incubation Time 1 h
Results The specificity of the reaction was further demonstrated by the loss of SENP3 phosphorylation as well as mTOR autophosphorylation upon the addition of the mTOR inhibitor Torin1.

Click to enlarge
Rating
Source Am J Pathol 2014 184(1), 214-29. Torin 1 purchased from Selleck
Method Western blot
Cell Lines Huh-7.5 cells
Concentrations 0-100 nM
Incubation Time 24 h
Results To investigate the relationship between ER stress and autophagy and IFN receptors, it first induced ER stress and autophagy response with the known inducers TG and Torin 1, respectively, in uninfected Huh-7.5 cells. It performed TG and Torin 1 titration experiments in HCV-infected and uninfected Huh-7.5 cells to compare IFNAR1 expression by Western blotting.

Click to enlarge
Rating
Source Biochem Biophys Res Commun 2014 10.1016/j.bbrc.2014.05.047. Torin 1 purchased from Selleck
Method Western blot
Cell Lines HepG2 cells
Concentrations 0.1 nM
Incubation Time 24 h
Results When autophagy was induced by treatment of mTOR inhibitors PP242 or Torin1, the amount of LC3-II increased.

Click to enlarge
Rating
Source Lim Hsiu Kim, Lina from National University of Singapore. Torin 1 purchased from Selleck
Method ELISA
Cell Lines C6
Concentrations 10 nM
Incubation Time 1 h
Results

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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  • Temsirolimus (CCI-779, NSC 683864)

    Temsirolimus (CCI-779, NSC 683864) is a specific mTOR inhibitor with IC50 of 1.76 μM in a cell-free assay.

  • INK 128 (MLN0128)

    INK 128 (MLN0128) is a potent and selective mTOR inhibitor with IC50 of 1 nM in cell-free assays; >200-fold less potent to class I PI3K isoforms, superior in blocking mTORC1/2 and sensitive to pro-invasion genes (vs Rapamycin). Phase 1.

  • Tacrolimus (FK506)

    Tacrolimus (FK506) is a 23-membered macrolide lactone, it reduces peptidyl-prolyl isomerase activity in T cells by binding to the immunophilin FKBP12 (FK506 binding protein) creating a new complex.

  • KU-0063794

    KU-0063794 is a potent and highly specific dual-mTOR inhibitor of mTORC1 and mTORC2 with IC50 of ~10 nM in cell-free assays; no effect on PI3Ks.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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