Product Categories

Ku-0063794

KU-0063794 is a potent and highly specific mTOR inhibitor for both mTORC1 and mTORC2 with IC50 ~10 nM.

Catalog No.S1226

3 Reviews 8 Product Citations

: Tel: +1-832-582-8158[email protected]

Ku-0063794 Chemical Structure
Molecular Weight: 465.54

Validation & Quality Control

Product Citations(8)

Cell Stem Cell, 2012, 10(2):210-7

Circ Res, 2010, 107(10), 1265-1274

Free Radic Biol Med, 2012, 53(5):1181-91

Br J Cancer, 2012, 106(8):1386-94

Biochem Pharmacol, 2012, 83(9), 1183-1194
J Biol Chem, 2011, 286(45):39450-6

Mol Cancer Res, 2012, 10(6), 821-833

PLoS One, 2012, 7(6), e39679

Customer Reviews(3) in vivo invitation

Quality Control & MSDS

Related Compound Libraries

Ku-0063794 is available in the following compound libraries:

Cambridge Cancer Compound Library(96-well) Chemical Library (96-well) Inhibitor Library (96-well) Kinase Inhibitor Library (96-well)

Product Information

Compare mTOR Inhibitors

Research Area
Research Area
Dr. Antonino Maria Sparta demonstrates a breakthrough in leukemia research by utilizing two inhibitors produced by Selleck Chemicals.

Cat.No.	Product Name	Solubility (25°C)
S1039	Rapamycin (Sirolimus)	<1 mg/mL	20 mg/mL	4 mg/mL
S1120	Everolimus (RAD001)	<1 mg/mL	30 mg/mL	7 mg/mL
S1009	BEZ235 (NVP-BEZ235)	<1 mg/mL	1 mg/mL	<1 mg/mL
S1555	AZD8055	2 mg/mL	93 mg/mL	15 mg/mL
S1038	PI-103	<1 mg/mL	24 mg/mL	<1 mg/mL
S2811	INK 128 (MLN0128)	<1 mg/mL	62 mg/mL	2 mg/mL
S2218	PP242	<1 mg/mL	62 mg/mL	18 mg/mL
S2827	Torin 1	<1 mg/mL	2 mg/mL	<1 mg/mL
S1022	Deforolimus (Ridaforolimus)	<1 mg/mL	198 mg/mL	<1 mg/mL
S1044	Temsirolimus (Torisel)	1 mg/mL	75 mg/mL	75 mg/mL
S1226	Ku-0063794	<1 mg/mL	16 mg/mL	<1 mg/mL
S1523	XL765 (SAR245409)	<1 mg/mL	15 mg/mL	<1 mg/mL
S2658	GSK2126458	<1 mg/mL	101 mg/mL	<1 mg/mL
S2624	OSI-027	<1 mg/mL	18 mg/mL	<1 mg/mL
S2743	PF-04691502	<1 mg/mL	14 mg/mL	<1 mg/mL
S2783	AZD2014	<1 mg/mL	38 mg/mL	<1 mg/mL
S2628	PF-05212384 (PKI-587)	<1 mg/mL	2 mg/mL	<1 mg/mL
S2696	GDC-0980 (RG7422)	<1 mg/mL	20 mg/mL	<1 mg/mL
S1266	WYE-354	<1 mg/mL	99 mg/mL	<1 mg/mL
S1360	GSK1059615	<1 mg/mL	2 mg/mL	<1 mg/mL
S2661	WYE-125132	<1 mg/mL	104 mg/mL	<1 mg/mL
S2668	WYE-687	<1 mg/mL	0.5 mg/mL	<1 mg/mL
S2699	CH5132799	<1 mg/mL	12 mg/mL	<1 mg/mL
S2817	Torin 2	<1 mg/mL	22 mg/mL	<1 mg/mL
S2238	Palomid 529	<1 mg/mL	81 mg/mL	<1 mg/mL
S2622	PP-121	<1 mg/mL	64 mg/mL	2 mg/mL
S2749	NVP-BGT226	<1 mg/mL	30 mg/mL	<1 mg/mL
S2689	WAY-600	<1 mg/mL	22 mg/mL	2 mg/mL

Product Description

Biological Activity Protocol Chemical Information Customer Reviews Product Citations Tech Support & FAQs

Biological Activity

Description	KU-0063794 is a potent and highly specific mTOR inhibitor for both mTORC1 and mTORC2 with IC50 ~10 nM.
Targets	mTORC1	mTORC2
IC50	~10 nM	~10 nM ^[1]
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]
In vivo
Clinical Trials
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

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.

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 Assay: ^[1]

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.

References

Chemical Information

Download Ku-0063794 SDF

Molecular Weight (MW)	465.54
Formula	C₂₅H₃₁N₅O₄
CAS No.	938440-64-3
Synonyms	N/A

Solubility (25°C) DMSO 16 mg/mL Water <1 mg/mL Ethanol <1 mg/mL
Storage	2 years -20°CPowder
	2 weeks4°Cin DMSO
	6 months-80°Cin DMSO

Chemical Name	(5-(2-((2R,6S)-2,6-dimethylmorpholino)-4-morpholinopyrido[2,3-d]pyrimidin-7-yl)-2-methoxyphenyl)methanol

Molarity Calculator Dilution Calculator Molecular Weight Calculator

Research Area

Customer Reviews (3)

Click to enlarge

Rating

Source

THE JOURNAL OF BIOLOGICAL CHEMISTRY, 2011. Ku-0063794 purchased from Selleck

Method

Western Blot

Cell Lines

mdMSC

Concentrations

2 uM

Incubation Time

Results

Here, the mTOR inhibitor KU0063794 (2 uM) reduced basal levels of phosphorylated Akt in mdMSC, leaving total Akt unchanged (Fig. 6A). KU0063794 prevented strain-induced phosphorylation of Akt at both Ser-473 and Thr-308… mTOR inhibition with KU0063794 disrupted insulin-induced phosphorylation of Akt at Ser-473 (Fig. 6D), but an increase in Akt phosphorylation at Thr-308 did occur.

Click to enlarge

Rating

Source

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

Method

MTT assays

Cell Lines

SUM149PT cells

Concentrations

0.001-10 µM

Incubation Time

72 h

Results

Ku-0063794 potently inhibited the survival of SUM149PT cells in a dose-dependent manner.

Click to enlarge

Rating

Source

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

Method

Western blot

Cell Lines

SMCs

Concentrations

10 µmol/L

Incubation Time

30 min

Results

We found that mTORC2 inhibitor Ku-0063794 blocked stretch- or IGF-1-induced SGK-1 phosphorylation. However, mTORC1 inhibitor rapamycin inhibited stretch-induced phosphorylation of p70S6K, but failed to inhibit SGK-1 phosphorylation, indicating that mTORC2 is a downstream signal of IGF-1R to activate SGK-1.

Product Citations (8)

Decoupling of tumor-initiating activity from stable immunophenotype in HoxA9-Meis1-driven AML. [Gibbs KD Jr, et al. Cell Stem Cell 2012;10(2):210-7]
PubMed: 22305570
The mechanical stress-activated serum-, glucocorticoid-regulated kinase 1 contributes to neointima formation in vein grafts. [Cheng J, et al. Circ Res 2010;107(10), 1265-1274]
PubMed: 20884880
Proline dehydrogenase is essential for proline protection against hydrogen peroxide induced cell death. [Natarajan SK, et al. Free Radic Biol Med 2012;53(5):1181-91]
PubMed: 22796327

The synergistic interaction of MEK and PI3K inhibitors is modulated by mTOR inhibition. [Haagensen EJ, et al. Br J Cancer 2012;106(8):1386-94]
PubMed: 22415236
Inhibition of tumor cell growth, proliferation and migration by X-387, a novel active-site inhibitor of mTOR. [Chen SM, et al. Biochem Pharmacol 2012;83(9), 1183-1194]
PubMed: 22305748
Mechanical regulation of GSK3β in mesenchymal stem cells is dependent on Akt serine-473 phosphorylation via mTOR complex 2. [Case N, et al. J Biol Chem 2011;286(45):39450-6]
PubMed: 21956113
Inhibition of mTORC1 kinase activates Smads 1 and 5 but not Smad8 in human prostate cancer cells, mediating cytostatic response to rapamycin. [Wahdan-Alaswad RS, et al. Mol Cancer Res 2012;10(6), 821-833]
PubMed: 22452883
mTOR is essential for the proteotoxic stress response, HSF1 activation and heat shock protein synthesis. [Chou SD, et al. PLoS One 2012;7(6), e39679]
PubMed: 22768106

Tech Support & FAQs

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3Monday–Friday 9:00 AM–5:00 PM (Central Time)

If you have any other enquiries, please leave a message.

* Indicates a Required Field

Related mTOR Inhibitors

BEZ235 (NVP-BEZ235)

BEZ235 (NVP-BEZ235) is a dual ATP-competitive PI3K and mTOR inhibitor of p110α, p110γ, p110δ and p110β with IC50 of 4 nM, 5 nM, 7 nM and 75 nM, respectively, and also inhibits ATR with IC50 of 21 nM.
Deforolimus (Ridaforolimus)

Deforolimus (Ridaforolimus, AP23573, MK-8669) is a selective mTOR inhibitor with IC50 of 0.2 nM.
PI-103

PI-103 is a potent, ATP-competitive PI3K inhibitor of DNA-PK, p110α, mTORC1, PI3KC2β, p110δ, mTORC2, p110β, and p110γ with IC50 of 2 nM, 8 nM, 20 nM, 26 nM, 48 nM, 83 nM, 88 nM and 150 nM, respectively.
WYE-354

WYE-354 is a potent, specific and ATP-competitive inhibitor of mTOR with IC50 of 5 nM.
GSK1059615

GSK1059615 is a novel and dual inhibitor of PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ and mTOR with IC50 of 0.4 nM, 0.6 nM, 2 nM, 5 nM and 12 nM, respectively.
XL765 (SAR245409)

XL765 is a dual inhibitor of mTOR/PI3k for mTOR, p110α, p110β, p110γ and p110δ with IC50 of 157 nM, 39 nM, 113 nM, 9 nM and 43 nM, respectively.
AZD8055

AZD8055 is a novel ATP-competitive inhibitor of mTOR with IC50 of 0.8 nM.
PP242

PP242 is a selective mTOR inhibitor with IC50 of 8 nM.
Palomid 529

Palomid 529 (P529) is a PI3K/Akt/mTOR inhibitor for VEGF-A and bFGF with IC50 of 10 nM and 30 nM, respectively.
PP-121

PP-121 is a multi-target inhibitor of PDGFR, Hck, mTOR, VEGFR2, Src and Abl with IC50 of 2 nM, 8 nM, 10 nM, 12 nM, 14 nM and 18 nM, respectively, and also inhibits DNA-PK with IC50 of 60 nM.

PI3K/Akt/mTOR Pathway Products

> Download

Choose Your Country or Region

Product Categories

Ku-0063794