| Field | Specification |
|---|---|
| Target | |
| CAS no. | |
| Applications | |
| Molecular weight | |
| Molecular formula | C25H31N5O4 |
| Purity | |
| SMILES | |
| Form | Solid |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Compound Overview
KU-0063794 is a potent, specific inhibitor of mTOR, blocking both the mTORC1 and mTORC2 complexes with an IC50 of 10 nM for each. It is supplied as a light yellow to yellow solid (C25H31N5O4, MW 465.54) at 99.67% purity.
Physical & Chemical Properties
| CAS Number | 938440-64-3 |
|---|---|
| Molecular Formula | C25H31N5O4 |
| Molecular Weight | 465.54 g/mol |
| Purity | 99.67% |
| Appearance | Solid |
| Color | Light yellow to yellow |
| SMILES | OCC1=CC(C2=CC=C3C(N=C(N=C3N4CCOCC4)N5C[C@@H](O[C@@H](C5)C)C)=N2)=CC=C1OC |
| Target | mTORC1, mTORC2 |
| Signaling Pathway | PI3K/Akt/mTOR |
| Solubility | In Vitro: DMSO: 16.67 mg/mL (35.81 mM; Requires sonication; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO) |
| Storage | Powder: -20°C, 3 years; 4°C, 2 years. In solvent: -80°C, 2 years; -20°C, 1 year. |
| Shipping | Room temperature in continental US; may vary elsewhere. |
Biological Activity
IC50 & Target[1]
|
mTORC1 10 nM (IC50) |
mTORC2 10 nM (IC50) |
Literature Cited
Sources cited in this description and in the In Vitro & In Vivo Data tab. Peer-reviewed publications that used this product are listed under References.
Safety
For Research Use Only. Not for use in diagnostic or therapeutic procedures, and not for human or veterinary use. Handle in accordance with the Safety Data Sheet and your institution's chemical hygiene plan.
In Vitro
| Solvent | Solubility | Notes |
|---|---|---|
| DMSO | 16.67 mg/mL (35.81 mM) | requires sonication; use freshly opened DMSO (absorbed moisture lowers solubility) |
Aliquot the stock solution and store it at -80°C (up to 2 years) or -20°C (up to 1 year); avoid repeated freeze-thaw cycles.
In Vivo
Choose the formulation that suits the animal model and route of administration; percentages are volume ratios of the final working solution. Start from a clear DMSO stock (see In Vitro above), add the co-solvents one at a time in the order listed, mixing after each addition, and prepare the working solution fresh on the day of dosing. If precipitation or phase separation occurs, gentle warming or sonication can help.
Protocol 1
| Composition | 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline |
|---|---|
| Result | ≥ 1.67 mg/mL (3.59 mM); clear solution |
| How to prepare | Gives a clear solution at ≥ 1.67 mg/mL (saturation not determined). For 1 mL of working solution: add 100 μL DMSO stock (16.7 mg/mL) to 400 μL PEG300; then 50 μL Tween-80; then 450 μL saline to bring the volume to 1 mL. Saline: dissolve 0.9 g sodium chloride in ddH2O and make up to 100 mL. |
Protocol 2
| Composition | 10% DMSO + 90% (20% SBE-β-CD in saline) |
|---|---|
| Result | ≥ 1.67 mg/mL (3.59 mM); clear solution |
| How to prepare | Gives a clear solution at ≥ 1.67 mg/mL (saturation not determined). For 1 mL of working solution: add 100 μL DMSO stock (16.7 mg/mL) to 900 μL 20% SBE-β-CD in saline. 20% SBE-β-CD in saline: dissolve 2 g SBE-β-CD powder in 10 mL saline until clear (4°C, store up to one week). |
Protocol 3
| Composition | 10% DMSO + 90% Corn Oil |
|---|---|
| Result | ≥ 1.67 mg/mL (3.59 mM); clear solution |
| How to prepare | Gives a clear solution at ≥ 1.67 mg/mL (saturation not determined). Use with caution if continuous dosing will exceed two weeks. For 1 mL of working solution: add 100 μL DMSO stock (16.7 mg/mL) to 900 μL corn oil. |
Data provided by the manufacturer.
In Vitro
Ku-0063794 is cell permeant and suppresses the activation and hydrophobic motif phosphorylation of Akt, S6K and SGK, though not of RSK (ribosomal S6 kinase; an AGC kinase that mTOR does not regulate). Ku-0063794 also inhibits phosphorylation of the T-loop Thr308 residue of Akt by PDK1 (3-phosphoinositide-dependent protein kinase-1). Ku-0063794 dephosphorylates the mTORC1 substrate 4E-BP1 (eukaryotic initiation factor 4E-binding protein 1) much more strongly than rapamycin does, even in mTORC2-deficient cells, suggesting that 4E-BP1 is phosphorylated by a form of mTOR distinct from mTORC1 or mTORC2. Ku-0063794 also suppresses cell growth and induces a G1-cell-cycle arrest[1]. Ku0063794 leaves nuclear phospho-Mst1-Thr-120 levels unchanged in LNCaP cell nuclei, whereas in C4-2 cell nuclei, Ku0063794 or CCI-779 increases phospho-Mst1-Thr-120 levels[2]. Combining GDC-0941 and KU0063794 inhibits phosphorylation of 4EBP1 and S6 in HCT116, DLD1 and HT29 cell lines to a similar extent as single agent NVP-BEZ235[3].
Data provided by the manufacturer. Numbered citations refer to the Literature Cited list in the product description.
Kinase Assay[1]
Freshly lyse HEK-293 cells in Hepes lysis buffer. Pre-clear the lysate (1-4 mg) by incubating with 5-20 μL of Protein G-Sepharose conjugated to pre-immune IgG. Next, incubate the lysate extracts with 5-20 μL of Protein G-Sepharose conjugated to 5-20 μg of anti-Rictor antibody, anti-Raptor antibody, or pre-immune IgG (all antibodies being covalently coupled to Protein G-Sepharose). Carry out immunoprecipitations for 1 hour at 4°C on a vibrating platform. Wash the immunoprecipitates four times with Hepes lysis buffer, then twice with Hepes kinase buffer. In the first two wash steps for Raptor immunoprecipitates used to phosphorylate S6K1, include 0.5mol/LNaCl in the buffer to ensure optimal kinase activity. Isolate GST-Akt1 from HEK-293 cells that were serum-deprived and incubated with PI-103 (1 μM for 1 hour). Purify GST-S6K1 from HEK-293 cells that were serum-deprived and incubated with rapamycin (0.1 μM for 1 hour). Start mTOR reactions by adding 0.1 mM ATP and 10 mM MgCl2, together with various concentrations of KU-0063794 plus either GST-Akt1 (0.5 μg) or GST-S6K1 (0.5 μg). Run the reactions for 30 minutes at 30°C on a vibrating platform, then stop them by adding SDS sample buffer. Filter the reaction mixtures through a 0.22-μM-poresize Spin-X filter, and analyze the samples by electrophoresis and immunoblotting with the indicated antibodies.
Cell Assay[1]
Treat cells with KU-0063794 for 24, 48, and 72 hours, changing the medium every 24 hours with freshly dissolved KU-0063794. To measure cell growth, wash cells once with PBS and fix in 4% (v/v) paraformaldehyde in PBS for 15 minutes. Following a single water wash, stain the cells with 0.1% Crystal Violet in 10% ethanol for 20 minutes and wash three times with water. Extract Crystal Violet from the cells using 0.5 mL of 10% (v/v) ethanoic (acetic) acid for 20 minutes. Dilute the eluate 1:10 in water and quantify absorbance at 590 nm. To assess cell cycle distribution, harvest cells by trypsinization, wash once in PBS, and re-suspend in ice-cold aq. 70% (v/v) ethanol. Wash cells twice in PBS plus 1% (w/v) BSA and stain for 20 minutes in PBS plus 0.1% (v/v) Triton X-100 supplemented with 50 g/mL propidium iodide and 50 g/mL RNase A. Determine DNA content of the cells with a FACSCalibur flow cytometer and CellQuest software. Acquire red fluorescence (585 nm) on a linear scale and use pulse width analysis to exclude doublets. Determine cell-cycle distribution with FlowJo software.
Data provided by the manufacturer. Numbered citations refer to the Literature Cited list in the product description.
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Distinct longevity mechanisms across and within species and their association with aging. Cell 2023 Jun 22;186(13):2929-2949.e20. PMID: 37269831
Identification and Application of Gene Expression Signatures Associated with Lifespan Extension. Cell Metab 2019 Sep 3;30(3):573-593.e8.
DYRK1B blockade promotes tumoricidal macrophage activity in pancreatic cancer. Gut 2024 Jun 4:gutjnl-2023-331854. PMID: 38834297
Intrinsic endothelial remodeling drives brain capillary repair. Neuron 2026 May 11:S0896-6273(26)00315-6. PMID: 42119560
Vimentin binds to a novel tumor suppressor protein, GSPT1-238aa, encoded by circGSPT1 with a selective encoding priority to halt autophagy in gastric carcinoma. Cancer Lett 2022 Oct 1:545:215826. PMID: 35839920
TOR signaling regulates liquid phase separation of the SMN complex governing snRNP biogenesis. Cell Rep 2021 Jun 22;35(12):109277. PMID: 34161763
CC-223, NSC781406, and BGT226 Exerts a Cytotoxic Effect Against Pancreatic Cancer Cells via mTOR Signaling. Front Pharmacol 2020 Nov 11:11:580407. PMID: 33343350
In Vitro and in Vivo Activity of mTOR Kinase and PI3K Inhibitors Against Leishmania donovani and Trypanosoma brucei. Molecules 2020 Apr 23;25(8):1980.
Inhibition of mTOR/S6K1/Gli1 signaling alleviates morphine-induced thermal hyperalgesia and tolerance. Mol Pain 2025 Jan-Dec:21:17448069251376198. PMID: 40968661
Selective ATP-competitive inhibitors of TOR suppress rapamycin-insensitive function of TORC2 in Saccharomyces cerevisiae. ACS Chem Biol 2012 Jun 15;7(6):982-7.
