1. PI3K/Akt/mTOR
    Autophagy
  2. mTOR
    Autophagy

Torin 1 

Cat. No.: HY-13003 Purity: 99.16%
Handling Instructions

Torin 1 is a potent inhibitor of mTOR with an IC50 of 3 nM. Torin 1 inhibits both mTORC1/2 complexes with IC50 values between 2 and 10 nM.

For research use only. We do not sell to patients.

Torin 1 Chemical Structure

Torin 1 Chemical Structure

CAS No. : 1222998-36-8

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Estimated Time of Arrival: December 31
50 mg USD 462 In-stock
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100 mg USD 768 In-stock
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Customer Review

    Torin 1 purchased from MCE. Usage Cited in: Haematologica. 2017 Apr;102(4):755-764.

    Knocking out of 4EBPs induces resistance to TORKi treatment. Ramos is transduced with CRISPR-CAS9 vectors targeting both 4EPB1 and 4EBP2 and immunoblotted with the indicated antibodies. 48 h after treatment with AZD8055 or Torin1, 4EBP1/2 double knock-out (Ramos-DKO) and control (Ramos-C) Ramos cells are immunoblotted with antibodies against MCL1 and BCL-XL

    Torin 1 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 May 22;9(6):604.

    AGS cells are pretreated with 25 and 50 nM Bortezomib for 1 h, and then incubated with 500 nM Torin 1 for 24 h. After pretreatment with PPI for 24 h in pH 7.4 or pH 6.5 condition, Rapamycin (1 μM) or Torin 1 (500 nM) is added for another 24 h. The protein level of SQSTM1 is measured by western blot analysis.

    Torin 1 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 May 22;9(6):604.

    Western blot analysis of ER stress-related proteins after PPI treatment for 48 h in both pH 7.4 and pH 6.5 conditions. Cells treated with Thapsigargin (TG, 0.5 and 1 μM) or Tunicamycin (Tu, 2.5, 5 and 10 μg/mL) for 24 h served as positive controls.

    Torin 1 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 May 22;9(6):604.

    AGS cells transfected with HA-tagged SQSTM1 plasmid, are treated with 25 μg/mL Cycloheximide (CHX) over a 240-min time period or treated with 100 μg/mL PPI for 48 h in pH 7.4 conditions, and then followed by 25 μg/mL CHX over a 240-min time period.

    Torin 1 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 May 22;9(6):604.

    Different concentrations of two classical proteasome inhibitors Bortezomib and MG132 are added. AGS cells are either untreated or treated with Bortezomib (25 nM) or MG132 (0.1 μM) for 24 h in the absence or presence of baf A1 (100 nM).

    Torin 1 purchased from MCE. Usage Cited in: Mol Cell Biol. 2017 Jun 29;37(14). pii: e00075-17.

    L-glutamine-responsive TORC1 activation in vitro. An in vitro kinase assay using semi-intact cells is performed. The reaction is performed with or without the indicated amount of Rapamycin or Torin-1. FLAG-tagged Tor1 and Myc-tagged Kog1 are detected as loading controls. The bar graph refers to the mean ratio of phosphorylated/total 4EBP1, normalized to the values for the DMSO-treated and glutamine-unadded samples.

    Torin 1 purchased from MCE. Usage Cited in: Cell Metab. 2018 Jan 9;27(1):118-135.e8.

    Inhibiting mTORC1 activity by intraperitoneally injecting DSS-treated C57BL/6 mice with Deforolimus or Torin 1 every other day also results in significant body weight loss compared with controls.

    Torin 1 purchased from MCE. Usage Cited in: Exp Cell Res. 2018 Jun 15;367(2):186-195.

    Torin1 and Rapamycin downregulate phospho-mTOR (Ser 2448), phospho-S6K1 (Thr 389), phospho-4EBP1 (Thr 37/46), Bcl-2 and upregulate Bax, cleaved caspase-3 in vitro cultured RSC96 cells.

    Torin 1 purchased from MCE. Usage Cited in: Exp Cell Res. 2018 Jun 15;367(2):186-195.

    MHY1485 prevents high glucose-caused reduced phospho-S6K1 (Thr 389) and cell apoptosis in RSC96 cells.

    Torin 1 purchased from MCE. Usage Cited in: J Virol. 2018 Jul 31;92(16). pii: e00600-18.

    HEK293T cells treated with Torin 1 (250 nM, 6 h) are used as the positive control. The cells are harvested at 30 h posttransfection and the lysates are analyzed with immunoblotting using antibodies specific for the indicated proteins.

    Torin 1 purchased from MCE. Usage Cited in: J Virol. 2018 Jul 31;92(16). pii: e00600-18.

    MARC-145 cells grown in six-well plates are treated with 0.5 mM Ars at 37°C for 1 h. The medium is changed to medium containing different concentrations of ISRIB and after 25 min, the cells are treated with puromycin, incubated at 37°C for 25 min, and lysed for western blotting analysis.

    Torin 1 purchased from MCE. Usage Cited in: Neurochem Res. 2018 Nov;43(11):2141-2154.

    Torin 1 (T) stimulation decreases the ratios of phospho-mTOR (Ser 2448)/ mTOR, phospho-S6K1 (Thr 389)/S6K1 and NGF/β-actin at all time points.

    Torin 1 purchased from MCE. Usage Cited in: Neurochem Res. 2018 Nov;43(11):2141-2154.

    MHY1485 (M) increases phospho-mTOR (Ser 2448), phosphoS6K1 (Thr 389) and NGF expression in RSC96 cells.

    Torin 1 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 Oct 9;9(10):1032.

    Western blot showing the cell cycle and autophagy-related proteins after 48 h of treatment with 200 μM Nitazoxanide (NTZ) and/or 500 nM Torin 1 or 30 mM Chloroquine (CQ).

    Torin 1 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 Oct 9;9(10):1032.

    Western blot showing SQSTM1 and LC3 levels in LN229 cells after treatment with 200 μM Nitazoxanide (NTZ) and 200 nM Bafilomycin A1 (BAF) for 48 h.

    View All mTOR Isoform Specific Products:

    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    Torin 1 is a potent inhibitor of mTOR with an IC50 of 3 nM. Torin 1 inhibits both mTORC1/2 complexes with IC50 values between 2 and 10 nM.

    IC50 & Target

    IC50: 2-10 nM (mTORC1), 2-10 nM (mTORC2), 3 nM (mTOR), 0.6 μM (ATM), 1 μM (DNA-PK), 1.8 μM (PI3K-α), 3 μM (hVPS34)[2]

    In Vitro

    Torin1 (250 nM) completely inhibits proliferation and causes a G1/S cell cycle arrest, and decreases cell size to a greater degree than 50 nM rapamycin in wild-type MEFs[1]. Torin1 has more than 800-fold selectivity between mTOR and PI3Kis, and is very selective relative to other PIKK family kinases with the exception of DNA-PK[2].

    In Vivo

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

    Solvent & Solubility
    In Vitro: 

    DMSO : 6.6 mg/mL (10.86 mM; Need ultrasonic and warming)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.6458 mL 8.2288 mL 16.4577 mL
    5 mM 0.3292 mL 1.6458 mL 3.2915 mL
    10 mM 0.1646 mL 0.8229 mL 1.6458 mL
    *Please refer to the solubility information to select the appropriate solvent.
    In Vivo:
    • 1.

      Torin 1 is dissolved in 100% N-methyl-2-pyrrolidone and subsequently diluted with PEG400 and water at the ratio of 1:2:2[3].

    • 2.

      Torin 1 is dissolved in NMP/PEG 400 (1:4)[4].

    References
    Kinase Assay
    [1]

    To produce soluble mTORC1, HEK-293T cell lines are generated that stably express N-terminally FLAG-tagged Raptor using vesicular stomatitis virus G-pseudotyped MSCV retrovirus. For mTORC2, HeLa cells are generated that stably express N-terminally FLAG-tagged Protor-1. 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 mol/L 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. 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.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Cell Assay
    [1]

    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.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Animal Administration
    [2]

    For pharmacodynamic experiments, torin 1 powder is first dissolved at 25 mg/mL in 100% N-methyl-2-pyrrolidone and then diluted 1:4 with sterile 50% PEG400 prior to injection. Six-week old male C57BL/6 mice are fasted overnight prior to drug treatment. The mice are treated with vehicle (for 10 hr) or 26 (20 mg/kg for 2, 6 or 10 hr) by IP injection, and then refed 1 h prior to sacrifice (CO2 asphyxiation). Tissues are collected and frozen on dry ice. The frozen tissue is thawed on ice and lysed by sonication in tissue lysis buffer (50 mM HEPES, pH 7.4, 40 mM NaCl, 2 mM EDTA, 1.5 mM sodium orthovanadate, 50 mM sodium fluoride, 10 mM sodium pyrophosphate, 10 mM sodium β-glycerophosphate, 0.1% SDS, 1.0% sodium deoxycholate and 1.0% Triton, supplemented with protease inhibitor cocktail tablets). The concentration of clear lysate is measured using the Bradford assay and samples are subsequently normalized by protein content and analyzed by SDS-PAGE and immunoblotting.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References
    Molecular Weight

    607.62

    Formula

    C₃₅H₂₈F₃N₅O₂

    CAS No.

    1222998-36-8

    SMILES

    O=C1N(C2=C(C=C1)C=NC3=CC=C(C=C32)C4=CC5=CC=CC=C5N=C4)C6=CC=C(C(C(F)(F)F)=C6)N7CCN(CC7)C(CC)=O

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Purity: 99.16%

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    Cat. No.: HY-13003