2. PI3K/Akt/mTOR Cell Cycle/DNA Damage Autophagy Apoptosis
  3. mTOR DNA-PK Autophagy Apoptosis
  4. Torin 2

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.

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Torin 2 Chemical Structure

Torin 2 Chemical Structure

CAS No. : 1223001-51-1

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Based on 16 publication(s) in Google Scholar

Torin 2 Related Antibodies

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  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

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[1][4]

mTOR

2.81 nM (IC50)

mTOR

0.25 nM (EC50, Cell Assay)

mTORC1

 

mTORC2

 

DNA-PK

0.5 nM (IC50)

p110γ

5.67 nM (IC50)

PI3K-C2β

24.5 nM (IC50)

PI3K-C2α

28.1 nM (IC50)

hVps34

8.58 nM (IC50)

PI3K

200 nM (EC50, Cell Assay)

ΡΙ4Κβ

18.3 nM (IC50)

Autophagy

 

Cellular Effect
Cell Line Type Value Description References
HCT-116 GI50
32.19 nM
Compound: Torin2
Growth inhibition of human HCT116 cells measured after 72 hrs by Celltiter-Glo assay
Growth inhibition of human HCT116 cells measured after 72 hrs by Celltiter-Glo assay
[PMID: 32911078]
HeLa IC50
< 10 nM
Compound: 15
Inhibition of N-terminally FLAG-tagged mTORC2 (unknown origin) expressed in human HeLa cells using S6K1 or Akt1 as substrate after 20 mins by immunoblotting assay
Inhibition of N-terminally FLAG-tagged mTORC2 (unknown origin) expressed in human HeLa cells using S6K1 or Akt1 as substrate after 20 mins by immunoblotting assay
[PMID: 29211480]
HeLa GI50
97.95 nM
Compound: Torin2
Growth inhibition of human HeLa cells measured after 72 hrs by Celltiter-Glo assay
Growth inhibition of human HeLa cells measured after 72 hrs by Celltiter-Glo assay
[PMID: 32911078]
MOLM-13 IC50
< 1 nM
Compound: Torin2
Antiproliferative activity against human MOLM13 cells by Cell-Titer Glo assay
Antiproliferative activity against human MOLM13 cells by Cell-Titer Glo assay
[PMID: 30370766]
MOLM-14 IC50
< 1 nM
Compound: Torin2
Antiproliferative activity against human MOLM14 cells by Cell-Titer Glo assay
Antiproliferative activity against human MOLM14 cells by Cell-Titer Glo assay
[PMID: 30370766]
MV4-11 IC50
< 1 nM
Compound: Torin2
Antiproliferative activity against human MV4-11 cells by Cell-Titer Glo assay
Antiproliferative activity against human MV4-11 cells by Cell-Titer Glo assay
[PMID: 30370766]
PC-3 EC50
200 nM
Compound: 3, Torin2
Inhibition of PI3Kalpha in human PC3 cells expressing Akt1 S473D mutant assessed as phosphorylation of Akt Thr308 by immunoblotting
Inhibition of PI3Kalpha in human PC3 cells expressing Akt1 S473D mutant assessed as phosphorylation of Akt Thr308 by immunoblotting
[PMID: 21322566]
RD CC50
0.04 μM
Compound: 1
Cytotoxicity against human RD cells incubated for 4 days by CellTiter-Glo luminescence assay
Cytotoxicity against human RD cells incubated for 4 days by CellTiter-Glo luminescence assay
[PMID: 31082764]
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].

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

Torin 2 Related Antibodies
In Vivo

Torin 2 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 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 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].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Molecular Weight

432.40

Formula

C24H15F3N4O
CAS No.
Appearance

Solid

Color

White to light yellow

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
Shipping

Room temperature in continental US; may vary elsewhere.
Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 1 year
-20°C 6 months
Solvent & Solubility
In Vitro: 

DMSO : 15.62 mg/mL (36.12 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened 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
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
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Concentration
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Volume
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Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 1.56 mg/mL (3.61 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.56 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (15.6 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% 1-Methyl-2-pyrrolidinone    90% PEG300

    Solubility: ≥ 2 mg/mL (4.63 mM); Clear solution

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

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(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
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% 
+
%
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%
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Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.65%

SDS (393 KB)

COA (253 KB) LCMS (93 KB)

Handling Instructions (2659 KB)
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

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 2.3127 mL 11.5634 mL 23.1267 mL 57.8168 mL
5 mM 0.4625 mL 2.3127 mL 4.6253 mL 11.5634 mL
10 mM 0.2313 mL 1.1563 mL 2.3127 mL 5.7817 mL
15 mM 0.1542 mL 0.7709 mL 1.5418 mL 3.8545 mL
20 mM 0.1156 mL 0.5782 mL 1.1563 mL 2.8908 mL
25 mM 0.0925 mL 0.4625 mL 0.9251 mL 2.3127 mL
30 mM 0.0771 mL 0.3854 mL 0.7709 mL 1.9272 mL
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Torin 2 Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Torin 21223001-51-1Torin2Torin-2mTORDNA-PKAutophagyApoptosisMammalian target of RapamycinDNA-dependent protein kinaseInhibitorinhibitorinhibit

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