1. PI3K/Akt/mTOR
    Cell Cycle/DNA Damage
    Autophagy
  2. mTOR
    DNA-PK
    Autophagy

Torin 2 

Cat. No.: HY-13002 Purity: 99.89%
Handling Instructions

Torin 2 is an mTOR inhibitor with EC50 of 0.25 nM for inhibiting cellular mTOR activity, and exhibits 800-fold selectivity over PI3K (EC50: 200 nM). Torin 2 also inhibits DNA-PK with an IC50 of 0.5 nM in the cell free assay. Torin 2 can suppress both mTORC1 and mTORC2.

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

Torin 2 Chemical Structure

Torin 2 Chemical Structure

CAS No. : 1223001-51-1

Size Price Stock Quantity
Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO USD 106 In-stock
Estimated Time of Arrival: December 31
5 mg USD 96 In-stock
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10 mg USD 132 In-stock
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50 mg USD 396 In-stock
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100 mg USD 588 In-stock
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200 mg USD 1008 In-stock
Estimated Time of Arrival: December 31
500 mg   Get quote  
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    Torin 2 purchased from MCE. Usage Cited in: Mol Cancer Ther. 2018 Oct 3. pii: molcanther.0260.2018.

    The mTORC1/2 inhibitor Torin2 suppresses protein synthesis in the EW8 and TC71 Ewing sarcoma cell lines.

    Torin 2 purchased from MCE. Usage Cited in: Mol Cancer Ther. 2018 Oct 3. pii: molcanther.0260.2018.

    EW8 cells are treated with 50 nM Gemcitabine in combination with increasing doses of Prexasertib, LY2603618, or MK-8776 for 6 hours. Cell lysates are then collected and blotted for p-CHK1- 296.

    Torin 2 purchased from MCE. Usage Cited in: Mol Cancer Ther. 2018 Oct 3. pii: molcanther.0260.2018.

    EW8 and TC71 cells are treated with Olaparib (5 μM) or U0126 (5 μM) for 24 hours and then protein synthesis is assessed using puromycin labeling.

    Torin 2 purchased from MCE. Usage Cited in: Mol Cancer Ther. 2018 Oct 3. pii: molcanther.0260.2018.

    Immunoblot assessing protein synthesis, using puromycin labeling, and CHK1 levels of cell lines treated with the protein translation inhibitor 4EGI-1.

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    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    Torin 2 is an mTOR inhibitor with EC50 of 0.25 nM for inhibiting cellular mTOR activity, and exhibits 800-fold selectivity over PI3K (EC50: 200 nM). Torin 2 also inhibits DNA-PK with an IC50 of 0.5 nM in the cell free assay. Torin 2 can suppress both mTORC1 and mTORC2.

    IC50 & Target

    EC50: 0.25 nM (cellular mTOR), 200 nM (cellular PI3K)[1]
    IC50: 2.81 nM (mTOR), 0.5 nM (DNA-pK), 5.67 nM (p110γ), 8.58 nM (hVPS34), 18.3 nM (PI4Kβ), 24.5 nM (PI3K-C2β), 28.1 nM (PI3K-C2α)[1]
    mTORC1, mTORC2[4]

    In Vitro

    Torin 2 is subject to further profiling against a panel of lipid kinases with IC50s of 2.81 nM, 0.5 nM, 5.67 nM, 8.58 nM, 18.3 nM, 24.5 nM and 28.1 nM for mTOR, DNA-pK, p110γ, hVPS34, PI4Kβ, PI3K-C2β and PI3K-C2α, respectively. Torin 2 (Torin2) possesses a 250 pM EC50 for inhibiting mTOR in cells while maintaining 800-fold cellular selectivity relative to inhibition of PI3K and most other protein kinases[1]. Torin 2 (Torin2) exhibits potent biochemical and cellular activity against PIKK family kinases including ATM (EC50 28 nM), ATR (EC50 35 nM) and DNA-PK (EC50 118 nM). Torin 2 potently inhibits T308 of Akt, a direct substrate of PDK1 and an indirect substrate of PI3Ks, with an EC50 of less than 10 nM[2]. Torin-2 can suppress both mTORC1 and mTORC2[4].

    In Vivo

    Torin 2 (Torin2) exhibits good bioavailability and exposure and can maintain strong inhibition of mTOR activity in lung and liver to at least six hours after a single dose of 20 mg/kg. Torin 2 is easier to produce on scale and exhibits improved pharmacokinetic properties which should enable it use in vivo experiments[1]. Torin 2 (Torin2) strongly suppresses pS6K(T389) and p4EBP1(T37/46) and partly suppresses pAkt(T308). Treatment of mice with AZD6244 at 25 mg/kg results in a profound inhibition of pERK. Combined administration of Torin 2 (40 mg/kg) and AZD6244 (25 mg/kg) demonstrates strong inhibition of all pharmacodynamics markers[2]. Treatment with Torin 2 (Torin2) and Rapamycin induces IL-6 secretion by astrocytes and may contribute to the reduction of mechanical hypersensitivity after SCI. Torin1 and Torin 2 treatment increases IL-6 mRNA, suggesting that the PI3K-mTOR pathway is a negative regulator of IL-6 expression in astrocytes. Importantly, Torin 2 treatment does not show any cell toxicity, as no signs of cell death are observed by TUNEL assay or by detection of cleaved-caspase 3 by western blotting[3].

    Solvent & Solubility
    In Vitro: 

    10 mM in DMSO

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.3127 mL 11.5634 mL 23.1267 mL
    5 mM 0.4625 mL 2.3127 mL 4.6253 mL
    10 mM 0.2313 mL 1.1563 mL 2.3127 mL
    *Please refer to the solubility information to select the appropriate solvent.
    References
    Cell Assay
    [2]

    HCT116 cells are treated with 100 nM Torin 2 or AZD8055 for 1 hour before they are thoroughly washed out by 3×PBS and 1×DMEM medium. Then cells are incubated in DMEM medium for indicated time before they are lysed and collected using M-PER. Protein concentrations are measured and equal amount of proteins are loaded. Experiments are repeated three times and one set of results[2].

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

    Animal Administration
    [1][3]

    Mice[1]
    Six-week old male C57BL/6 mice are fasted overnight prior to Torin 2 treatment. The mice are treated with vehicle (for 10 h) or Torin 2 (20 mg/kg for 6h) via oral gavage and then re-fed 1 h prior to sacrifice (CO2 asphyxiation). Liver and lung 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.
    Rats[3]
    Female rats (220 g) are group-housed 4 animals per cage, and kept on a 12-hour light/dark cycle with food and water ad libitum. Spinal cord injury is done with the Keck Center for Neurosciences impactor using a 10 g weight dropped from a height of 25 mm onto the dorsal surface of the exposed spinal cord. After recording BBB scores, withdrawal thresholds evoked by touch stimulus, and body weights for the first week post-injury, animals are divided into 5 treatment groups: naïve (N=4), sham (N=6), vehicle (N=6), Torin 2 (N=6), and Torin 2+Rapamycin (N=8). Torin 2 alone (4 mg/kg) or in combination with Rapamycin (1.5 mg/kg) is administered orally by gavage once a day starting at day 15 after injury and ending at day 29. In sham operated rats only the laminectomy is performed.

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

    References
    Molecular Weight

    432.40

    Formula

    C₂₄H₁₅F₃N₄O

    CAS No.

    1223001-51-1

    SMILES

    O=C1N(C2=C(C=C1)C=NC3=CC=C(C=C32)C4=CC=C(N=C4)N)C5=CC=CC(C(F)(F)F)=C5

    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: 99.89%

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    Product Name:
    Torin 2
    Cat. No.:
    HY-13002
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    Torin 2

    Cat. No.: HY-13002