KU-0063794

Catalog No.S1226

KU-0063794 Chemical Structure

Molecular Weight(MW): 465.54

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.

Size Price Stock Quantity  
In DMSO USD 180 In stock
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4 Customer Reviews

  • mTORC2-mediated stretch-induced SGK-1 phosphorylation. SMCs infected with Ad-SGK-1 were pretreated with PPP (10 μmol/L), rapamycin (100 nmol/L),or Ku-0063794 (10 μmol/L) for 30 minutes and then were subjected to stretch or IGF-1 (10 ng/mL) for 30 minutes.

     

     

    Circ Res 2010 107, 1265-1274. KU-0063794 purchased from Selleck.

    Effects of PI3K and mTOR inhibitors on IGF1R and AKT signaling in RMS cells. Rh41 cells were treated with 0.3 uM PI3K inhibitor BKM120, 0.3 uM mTOR inhibitor KU0063794 for the indicated time. Lysates were made and analyzed for p-AKT, S6RP and IGF1R.

    Oncogene 2013 10.1038/onc.2013.509. KU-0063794 purchased from Selleck.

  •  

    mTORC2 is required for mechanical activation of Akt. A, immunoblots of mdMSC subjected to strain for 45 min following treatment with the mTOR inhibitor KU0063794 (2uM). B, densitometric analysis of phosphorylated GSK3 (Ser-9) normalized to total GSK3. D, immunoblots of mdMSC treated with KU0063794 and then stimulated with insulin (50 nM) for 60 min.

    J Biol Chem 2011 286, 39450-6. KU-0063794 purchased from Selleck.

    For MTT assays, cells (2,000 ~ 5,000 cells/well) were subcultured into 96-well plates according to their growth properties. Cell proliferation was assayed at 72 hr after treatment of KU-0063794 by adding 20 μl of 5 mg/ml 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution per 100 μl of growth medium. After incubating for 3-4 h at 37°C, the media were removed and 150 µl/well of MTT solvent (either absolute DMSO or isopropanol containing 4 μM HCl and 0.1% Nonidet-40) was added to dissolve the formazan. The absorbance of each well was measured by ELx808 (BioTek, Winooski, VT) or Wallac Victor2 (Perkin-Elmer Life Sciences, Boston, MA) Microplate Reader. Viable cells are presented as percent of control, vehicle-treated cells.

     

     

    Dr. Yong-Weon Yi from Georgetown University Medical Center. KU-0063794 purchased from Selleck.

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Biological Activity

Description 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.
Targets
mTORC1 [1]
(Cell-free assay)
mTORC2 [1]
(Cell-free assay)
~10 nM ~10 nM
In vitro

Compared with the mTOR inhibitor PP242, KU-0063794 exhibits higher specificity for mTOR, as being inactive against PI3Ks or 76 other kinases. In HEK-293 cells, KU-0063794 at 30 nM is sufficient to rapidly ablate S6K1 activity by blocking the phosphorylation of the hydrophobic motif (Thr389) and subsequently the phosphorylation of the T-loop residue (Thr229). In case of IGF1 stimulation of serum-starved HEK-293 cells, 300 nM of KU-0063794 is needed to inhibit the S6K1 activity by ~90%. KU-0063794 at 100-300 nM also completely inhibits the amino-acid-induced phosphorylation of S6K1 and S6 protein. Similar to S6K1, KU-0063794 inhibits the phosphorylation of mTORC1 at Ser2448 and mTORC2 at Ser2481 in a dose-dependent and time-dependent manner. In the presence of serum or following IGF1 stimulation, KU-0063794 induces a dose-dependent inhibition of the activity and phosphorylation of Akt at Ser473 and unexpected Thr308 as well as the phosphorylation of the Akt substrates PRAS40 at Thr246, GSK3α/GSK3β at Ser21/Ser9 and Foxo-1/3a at Thr24/Thr32. KU-0063794 but not rapamycin inhibits SGK1 activity and Ser422 phosphorylation as well as its physiological substrate NDGR1 in a dose-dependent manner, to the same extent as S6K1 and Akt phosphorylation, whereas KU-0063794 dose not inhibit phorbol ester induced ERK or RSK phosphorylation and RSK activation. Compared with rapamycin, KU-0063794 exhibits more significant potency to induce the complete dephosphorylation of 4E-BP1 at Thr37, Thr46 and Ser65. KU-0063794 inhibits cell growth of both wild-type and mLST8-deficient MEFs and induces a G1 cell cycle arrest, more significantly than rapamycin. [1]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HepG2  M{\ic2NmdGxiVnnhZoltcXS7IFHzd4F6 MmWyNE4y6oDVNUFCpO69VQ>? M4DsSVczKGh? Mnfs[IVkemWjc3XzJINmdGxidnnhZoltcXS7IHnuJIEh\G:|ZTDk[ZBmdmSnboSgcYFvdmW{ M3\Yb|I3Ojd6OEG5
HepG2  NHe2[I5Ed2yxbomgSo9zdWG2aX;uJGF{e2G7 MYCx5qCUPTEEoN88US=> M2TBTlExKGR? MYPk[YNz\WG|ZYOgeIhmKG63bXLldkBw\iC4aXHicIUhUGWyR{KgZ49td26rZYRCpJNq\26rZnnjZY51dHl? NUT1Zm9bOjZ{N{i4NVk>
HepG2  M2jqWmFxd3C2b4Ppd{BCe3OjeR?= M3fOV|AvOeLCk{WwxsDPxE1? MWm0PEBp MkTObY5lfWOnczDhdI9xfG:|aYOgbY4h[SCmb4PlJIRmeGWwZHXueEBu[W6wZYK= NX7INFFDOjZ{N{i4NVk>
HepG2  NI\uNXJHfW6ldHnvckBCe3OjeR?= Mn\ZOU8yOCEQvF2= NHfveI0zPCCq MkG5[JJidWG2aXPhcIx6KGmwaHnibZR{KHCqb4PwbI9zgWyjdHnvckBw\iCDS2SgZZQhW2W{LUS3Ny=> NUfOT4t2OjZ{N{i4NVk>
HepG2  M1nSN2Z2dmO2aX;uJGF{e2G7 MlPYOU8yOCEQvF2= MnTmNlQhcA>? MlLC[I94dnKnZ4XsZZRmeyC2aHWgcIV3\Wy|IFjJSlHPuSCjbnSgZ5lkdGmwIFSxxsA> MUeyOlI4QDhzOR?=
HepG2  MWfGeY5kfGmxbjDBd5NigQ>? MWWwMlHjiJN3MNMg{txO M2jXXFI1KGh? Mmn3bY5lfWOnczDwOlIh\G:5boLl[5Vt[XSrb36sJGJm[2yrbj2xJIV5eHKnc4Ppc44h[W6mIFzDN2IuUSC2bzDMR|NDNUmLIHPvcpZmenOrb36gbY4h[SCmb4PlJIRmeGWwZHXueEBu[W6wZYK= MnfWNlYzPzh6MUm=
HepG3  M{HNNWZ2dmO2aX;uJGF{e2G7 NHzhcmIxNjIkgKO1NOKh|ryP MmjGOFghcA>? MlywbY5lfWOnczDj[YxtKGG3dH;wbIFogSCrbjDhJIRwe2ViZHXw[Y5l\W62IH3hco5meg>? M2PU[|I3Ojd6OEG5
AGS M2LnNmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MUfJR|UxRTF3LkCgxtEhOi57MTFOwG0> MVqyOFU6PzR5OB?=
HGC27 MUHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MWrJR|UxRTF3LkCgxtEhPC56MjFOwG0> NXfJe206OjR3OUe0O|g>
MKN45 MmGxS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NELSRYJKSzVyPUCuPFIhyrFiMD6wNUDPxE1? NYHsV3JtOjR3OUe0O|g>
NUGC4 NFewUnRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NFTKSXdKSzVyPUKuPVMhyrFiMD6zNUDPxE1? NHLxd2MzPDV7N{S3PC=>
PC9 NGrDZZVIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M4fy[VczKGh? NHjjV2hKSzVyPUGwMlE2yrFyLk[yJI5O MW[yN|g4PDh6MB?=
PC9GR NXPK[G9zT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= Mn\BO|IhcA>? M2C2e2lEPTB;Nj6yNeKyOS5|MDDuUS=> MkDsNlM5PzR6OEC=
H1650 MmDBS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? Ml\5O|IhcA>? MkmzTWM2OD15Lk[xxtExNjZ{IH7N MUWyN|g4PDh6MB?=
H1975 M{DXXWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NFLLc4Y4OiCq NGPaTYtKSzVyPUGxMlE2yrFyLkmzJI5O MUGyN|g4PDh6MB?=
PC9 MVXGeY5kfGmxbjDBd5NigQ>? MnS1NVAvOTVibl2= MXS3NkBp M33Fc4lvcGmkaYTzJI1VV1JicHjvd5Bpd3K7bHH0bY9vKHO2YYT1d:Kh NIH1UlkzOzh5NEi4NC=>
PC9GR MX\GeY5kfGmxbjDBd5NigQ>? NH3STlU3NjJzIH7N MYG3NkBp MYrpcohq[mm2czDtWG9TKHCqb4PwbI9zgWyjdHnvckB{fGG2dYRCpC=> M3j5[FI{QDd2OEiw
H1650 NVn5NHRsTnWwY4Tpc44hSXO|YYm= MUO3MlYyKG6P NXjUZndPPzJiaB?= MUHpcohq[mm2czDtWG9TKHCqb4PwbI9zgWyjdHnvckB{fGG2dYRCpC=> MYCyN|g4PDh6MB?=
H1975 MofWSpVv[3Srb36gRZN{[Xl? MV6xNU4yPSCwTR?= NGHMSpc4OiCq MkX2bY5pcWKrdIOgcXRQWiCyaH;zdIhwenmuYYTpc44he3SjdIXzxsA> NF7FUoIzOzh5NEi4NC=>
PC9 NYHibplZTnWwY4Tpc44hSXO|YYm= M1\nS|ExNjF3IH7N MlXVO|IhcA>? M{HCdolvcGmkaYTzJJBpd3OyaH;yfYxifGmxbjDv[kBxPzCVNlu= NHLGTYIzOzh5NEi4NC=>
PC9GR MkDHSpVv[3Srb36gRZN{[Xl? MXW2MlIyKG6P MkTzO|IhcA>? M1zOdYlvcGmkaYTzJJBpd3OyaH;yfYxifGmxbjDv[kBxPzCVNlu= NUKzd2FqOjN6N{S4PFA>
H1650 M1HzTmZ2dmO2aX;uJGF{e2G7 NGXxO|A4NjZzIH7N MU[3NkBp MonpbY5pcWKrdIOgdIhwe3Cqb4L5cIF1cW:wIH;mJJA4OFN4Sx?= NUPudXV[OjN6N{S4PFA>
H1975 NIL5Z3pHfW6ldHnvckBCe3OjeR?= M{\KflEyNjF3IH7N NV\wbHFLPzJiaB?= NFPNUpdqdmirYnn0d{BxcG:|cHjvdplt[XSrb36gc4YheDdyU{\L NXi3W41yOjN6N{S4PFA>
LNCaP NWeyOVlMS2WubDDWbYFjcWyrdImgRZN{[Xl? MWKwMVExKM7:TR?= M3O5TlI1KGkEoB?= M1;mWYRm[3KnYYPld{Bk\WyuII\pZYJqdGm2eTDpckBiKGSxc3Wg[IVx\W6mZX70JI1idm6nch?= MXmyN|g1ODZyNR?=
PC-3 MXLD[YxtKF[rYXLpcIl1gSCDc4PhfS=> Mmn1NE0yOCEQvF2= M4PaT|I1KGkEoB?= MXfk[YNz\WG|ZYOgZ4VtdCC4aXHibYxqfHliaX6gZUBld3OnIHTldIVv\GWwdDDtZY5v\XJ? NWLZTJRSOjN6NEC2NFU>
MDA-MB-468  NG\teGhE\WyuIG\pZYJqdGm2eTDBd5NigQ>? MX:wMVExKM7:TR?= MYSyOEBpyqB? NILzO2Rl\WO{ZXHz[ZMh[2WubDD2bYFjcWyrdImgbY4h[SCmb4PlJIRmeGWwZHXueEBu[W6wZYK= M1qxRVI{QDRyNkC1
LNCaP MoDnSpVv[3Srb36gRZN{[Xl? NUHTV3BvOjBy4pETPFAxKG6P M4rVZlI1KGkEoB?= M{i2XIRm[3KnYYPld{B1cGVicHjvd5Bpd3K7bHH0bY9vKGyndnXsJI9nKHB5MGO2T{BqdiCjIHTvd4Uh\GWyZX7k[Y51KG2jbn7ldi=> MlrXNlM5PDB4MEW=
PC-3 M1\tcGZ2dmO2aX;uJGF{e2G7 NXPo[FRjOjBy4pETPFAxKG6P NEi0TWozPCCqwrC= NUW5cJlJ\GWlcnXhd4V{KHSqZTDwbI9{eGixconsZZRqd25ibHX2[Ywhd2ZicEewV|ZMKGmwIHGg[I9{\SCmZYDlcoRmdnRibXHucoVz NV2wTY5rOjN6NEC2NFU>
MDA-MB-468  MlfXSpVv[3Srb36gRZN{[Xl? M{e2O|IxOOLCk{iwNEBvVQ>? M4PlNVI1KGkEoB?= NWrWV29J\GWlcnXhd4V{KHSqZTDwbI9{eGixconsZZRqd25ibHX2[Ywhd2ZicEewV|ZMKGmwIHGg[I9{\SCmZYDlcoRmdnRibXHucoVz M1;oWVI{QDRyNkC1
Caki-1  NELid4NHfW6ldHnvckBCe3OjeR?= MYKxNFAuOjByMDDuUS=> Mm\SNVAuOThyIH3pci=> NFnwc5pFVVOR MV\pcohq[mm2czDic5RpKG2WT2LDNUBidmRibWTPVmMzKGG|IHnu[Ilk[XSnZDDifUB1cGViZHXjdoVie2ViaX6gdIhwe3Cqb4L5cIF1cW:wIH;mJIRwf26|dILlZY0h\W[oZXP0c5J{ MmXKNlM{PDl7OEm=
786-O NXf3ToplTnWwY4Tpc44hSXO|YYm= M1r2clExOC1{MECwJI5O NIHF[JgyOC1zOECgcYlv NIfjdlZFVVOR NWfjcVczcW6qaXLpeJMh[m:2aDDtWG9TSzFiYX7kJI1VV1KFMjDhd{BqdmSrY3H0[YQh[nlidHjlJIRm[3KnYYPlJIlvKHCqb4PwbI9zgWyjdHnvckBw\iCmb4fud5Rz\WGvIHXm[oVkfG:{cx?= MUeyN|M1QTl6OR?=
Caki-1  MnjzR4VtdCCYaXHibYxqfHliQYPzZZk> MoD1N|AxNTRyMECgcm0> MYKyOE06PiCq Ml\USG1UVw>? MYrzeZBxemW|c3XzJJRp\SClZXzsJJZq[WKrbHn0fUBqdiCkb4ToJJRqdWViYX7kJIRwe2ViZHXw[Y5l\W62IH3hco5meg>? M2jURlI{OzR7OUi5
786-O NEHOZ3FE\WyuIG\pZYJqdGm2eTDBd5NigQ>? MX2zNFAuPDByMDDuUS=> MWmyOE06PiCq NHPscGVFVVOR MVvzeZBxemW|c3XzJJRp\SClZXzsJJZq[WKrbHn0fUBqdiCkb4ToJJRqdWViYX7kJIRwe2ViZHXw[Y5l\W62IH3hco5meg>? NXqzXIl6OjN|NEm5PFk>
Caki-1  MVHGeY5kfGmxbjDBd5NigQ>? NHr5d4kzKML3TR?= NHj6SoM4OiCq MVvEUXNQ MUjpcoR2[2W|IFexJINmdGxiY4njcIUh[XK{ZYP0JIFv\CCjdYTvdIhi\3l? M2PTUlI{OzR7OUi5
786-O NXnHWZQ4TnWwY4Tpc44hSXO|YYm= M4nHV|IhyrWP M3XwVlczKGh? M1L1cGROW09? M3LWVIlv\HWlZYOgS|Eh[2WubDDjfYNt\SCjcoLld5Qh[W6mIHH1eI9xcGGpeR?= M4rMNVI{OzR7OUi5

... Click to View More Cell Line Experimental Data

In vivo Ku0063794 inhibits tumor growth and mTOR signaling in a preclinical renal cell carcinoma model. However, Ku0063794 was not more effective than temsirolimus in the animal study. A possible explanation for lack of greater activity in vivo for Ku0063794 is that temsirolimus has important effects on the tumor microenvironment. Temsirolimus decreased angiogenesis in the xenograft tumors while Ku0063794 did not. Temsirolimus treated tumors expressed less VEGF and PDGF than Ku0063794 treated tumors, thus stimulating less angiogenesis[2].

Protocol

Kinase Assay:

[1]

+ Expand

mTOR complexes kinase assays:

HEK-293 cells are freshly lysed in Hepes lysis buffer. Lysate (1-4 mg) is pre-cleared by incubating with 5-20 μL of Protein G-Sepharose conjugated to pre-immune IgG. The lysate extracts are then incubated with 5-20 μL of Protein G-Sepharose conjugated to 5-20 μg of either anti-Rictor or anti-Raptor antibody, or pre-immune IgG. All antibodies are covalently conjugated to Protein G-Sepharose. Immunoprecipitations are carried out for 1 hour at 4 °C on a vibrating platform. The immunoprecipitates are washed four times with Hepes lysis buffer, followed by two washes with Hepes kinase buffer. For Raptor immunoprecipitates used for phosphorylating S6K1, for the initial two wash steps the buffer includes 0.5 M NaCl to ensure optimal kinase activity. GST-Akt1 is isolated from serum-deprived HEK-293 cells incubated with PI-103 (1 μM for 1 hour). GST-S6K1 is purified from serum-deprived HEK-293 cells incubated with rapamycin (0.1 μM for 1 hour). mTOR reactions are initiated by adding 0.1 mM ATP and 10 mM MgCl2 in the presence of various concentrations of KU-0063794 and GST-Akt1 (0.5 μg) or GST-S6K1 (0.5 μg). Reaction are carried out for 30 minutes at 30 °C on a vibrating platform and stopped by addition of SDS sample buffer. Reaction mixtures are then filtered through a 0.22-μm-poresize Spin-X filter and samples are subjected to electrophoresis and immunoblot analysis with the indicated antibodies.
Cell Research:

[1]

+ Expand
  • Cell lines: Wild-type and mLST8 deficient MEFs
  • Concentrations: Dissolved in DMSO, final concentration ~3 μM
  • Incubation Time: 24, 48, and 72 hours
  • Method:

    Cells are treated with KU-0063794 for 24, 48, and 72 hours, and the medium is changed every 24 hours with freshly dissolved KU-0063794. For the measurement of cell growth, cells are washed once with PBS, and fixed in 4% (v/v) paraformaldehyde in PBS for 15 minutes. After washing once with water, the cells are stained with 0.1% Crystal Violet in 10% ethanol for 20 minutes and washed three times with water. Crystal Violet is extracted from cells with 0.5 mL of 10% (v/v) ethanoic (acetic) acid for 20 minutes. The eluate is then diluted 1:10 in water and absorbance at 590 nm is quantified. For the assessment of cell cycle distribution, cells are harvested by trypsinization, washed once in PBS, and re-suspended in ice-cold aq. 70% (v/v) ethanol. Cells are washed twice in PBS plus 1% (w/v) BSA and stained for 20 minutes in PBS plus 0.1% (v/v) Triton X-100 containing 50 g/mL propidium iodide and 50 g/mL RNase A. The DNA content of cells is determined using a FACSCalibur flow cytometer and CellQuest software. Red fluorescence (585 nm) is acquired on a linear scale, and pulse width analysis is used to exclude doublets. Cell-cycle distribution is determined using FlowJo software.


    (Only for Reference)
Animal Research:

[2]

+ Expand
  • Animal Models: Nu/Nu nude mice
  • Formulation: one part DMSO and then diluted (200 µg/100 µl) with 4 parts PEG1500 (50% (w/v) in 75 mM Hepes, pH 8.0
  • Dosages: 8 mg/kg
  • Administration: i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 16 mg/mL (34.36 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents individually and in order:
30% PEG400+0.5% Tween80+5% propylene glycol
30 mg/mL

* 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 Information

Molecular Weight 465.54
Formula

C25H31N5O4

CAS No. 938440-64-3
Storage powder
Synonyms N/A

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID