KU-0063794

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.

CAS No. 938440-64-3

サイズ	価格(税別)	在庫状況
10mM (1mL in DMSO)	JPY 31800	国内在庫あり
5mg	JPY 16900	国内在庫あり
10mg	JPY 30200	国内在庫あり
50mg	JPY 80000	国内在庫あり

代表番号: 045-509-1970\|電子メール：[email protected] よく尋ねられる質問

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製品安全説明書

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KU-0063794関連製品

関連ターゲット	mTORC1 mTORC2	もっと見る
関連化合物ライブラリー	Kinase Inhibitor Library PI3K/Akt Inhibitor Library Apoptosis Compound Library Cell Cycle compound library NF-κB Signaling Compound Library	もっと見る

シグナル伝達経路

mTORシグナル伝達経路

mTOR阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
H1650	Function Assay	7.61 nM	72 h	inhibits phosphorylation of p70S6K	23874880
PC9GR	Function Assay	6.21 nM	72 h	inhibits phosphorylation of p70S6K	23874880
PC9	Function Assay	10.15 nM	72 h	inhibits phosphorylation of p70S6K	23874880
H1975	Function Assay	11.15 nM	72 h	inhibits mTOR phosphorylation status	23874880
H1650	Function Assay	7.61 nM	72 h	inhibits mTOR phosphorylation status	23874880
PC9GR	Function Assay	6.21 nM	72 h	inhibits mTOR phosphorylation status	23874880
PC9	Function Assay	10.15 nM	72 h	inhibits mTOR phosphorylation status	23874880
HepG3	Function Assay	0.1–50 μM	48 h	induces cell autophagy in a dose dependent manner	26278819
HepG2	Function Assay	0.1–50 μM	24 h	induces p62 downregulation, Beclin-1 expression and LC3B-I to LC3B-II conversion in a dose dependent manner	26278819
HepG2	Function Assay	5/10 μM	24 h	downregulates the levels HIF1α and cyclin D1	26278819
HepG2	Function Assay	5/10 μM	24 h	dramatically inhibits phosphorylation of AKT at Ser-473	26278819
HepG2	Apoptosis Assay	0.1–50 μM	48 h	induces apoptosis in a dose dependent manner	26278819
HepG2	Colony Formation Assay	1–50 μM	10 d	decreases the number of viable HepG2 colonies significantly	26278819
HepG2	Cell Viability Assay	0.1–50 μM	72 h	decreases cell viability in a dose dependent manner	26278819
H1975	Function Assay	11.15 nM	72 h	inhibits phosphorylation of p70S6K	23874880
LNCaP	Cell Viability Assay	0-10 μM	24 h	decreases cell viability in a dose dependent manner	23840605
PC-3	Cell Viability Assay	0-10 μM	24 h	decreases cell viability in a dose dependent manner	23840605
MDA-MB-468	Cell Viability Assay	0-10 μM	24 h	decreases cell viability in a dose dependent manner	23840605
LNCaP	Function Assay	200–800 nM	24 h	decreases the phosphorylation level of p70S6K in a dose dependent manner	23840605
PC-3	Function Assay	200–800 nM	24 h	decreases the phosphorylation level of p70S6K in a dose dependent manner	23840605
MDA-MB-468	Function Assay	200–800 nM	24 h	decreases the phosphorylation level of p70S6K in a dose dependent manner	23840605
Caki-1	Function Assay	100-2000 nM	10-180 min	inhibits both mTORC1 and mTORC2 as indicated by the decrease in phosphorylation of downstream effectors	23349989
786-O	Function Assay	100-2000 nM	10-180 min	inhibits both mTORC1 and mTORC2 as indicated by the decrease in phosphorylation of downstream effectors	23349989
Caki-1	Cell Viability Assay	300-4000 nM	24-96 h	suppresses the cell viability in both time and dose dependent manner	23349989
786-O	Cell Viability Assay	300-4000 nM	24-96 h	suppresses the cell viability in both time and dose dependent manner	23349989
Caki-1	Function Assay	2 µM	72 h	induces G1 cell cycle arrest and autophagy	23349989
786-O	Function Assay	2 µM	72 h	induces G1 cell cycle arrest and autophagy	23349989
HBCx-10	Function assay	5 mg/kg		Potentiation of irinotecan-induced tumor regression against human HBCx-10 cells xenografted in immunocompromised mouse at 5 mg/kg, po qd administered on days 1 to 3 of weekly cycle	19762236
PC3	Function assay	100 mg/kg		Inhibition of Akt phosphorylation at Ser473 in PTEN-deficient human PC3 cells xenograft mouse model at 100 mg/kg, po single dose measured up to 8 hrs	29211480
PC3	Antitumor assay	30 mg/kg		Antitumor activity against human PC3 cells xenograft mouse model assessed as inhibition of tumor growth at 30 mg/kg, po bid in presence of 1-aminobenzotriazole	29211480
PC3	Antitumor assay	60 mg/kg		Antitumor activity against human PC3 cells xenograft mouse model assessed as inhibition of tumor growth at 60 mg/kg, po bid in presence of 1-aminobenzotriazole	29211480
SW620	Function assay	20 mg/kg		Potentiation of irinotecan-induced tumor regression against human SW620 cells xenografted in immunocompromised mouse at 20 mg/kg, po qd administered on days 2 to 4 of weekly cycle	29211480
H1975	Growth Inhibition Assay		72 h	IC50=11.15±0.93 nM	23874880
H1650	Growth Inhibition Assay		72 h	IC50=7.61±0.62 nM	23874880
PC9GR	Growth Inhibition Assay		72 h	IC50=6.21±1.30 nM	23874880
PC9	Growth Inhibition Assay		72 h	IC50=10.15±0.62 nM	23874880
HEK293	Function assay		2 hrs	Inhibition of recombinant FLAG-tagged mTOR expressed in HEK293 cells using biotinylated p70S6K substrate after 2 hrs by alphascreen competition assay, IC50=0.003μM	19762236
U87MG	Function assay		2 hrs	Inhibition of mTORC1 in human U87MG cells assessed as phosphorylated S6 ribosomal protein (Ser235/236) level after 2 hrs by Western blotting, IC50=0.1μM	19762236
U87MG	Function assay		2 hrs	Inhibition of mTORC2 in human U87MG cells assessed as phosphorylated AKT (Ser473) level after 2 hrs by Western blotting, IC50=0.15μM	19762236
T47D	Antiproliferative assay		120 hrs	Antiproliferative activity against human T47D cells after 120 hrs by SRB assay, GI50=0.35μM	19762236
MDA-MB-468	Function assay		2 hrs	Inhibition of mTORC2 in human MDA-MB-468 cells assessed as reduction of AKT phosphorylation at Ser473 after 2 hrs, IC50=0.24μM	23375793
MDA-MB-468	Function assay		2 hrs	Inhibition of mTORC1 in human MDA-MB-468 cells assessed as reduction of pS6 phosphorylation at Ser235/236 after 2 hrs, IC50=0.66μM	23375793
HEK293	Function assay		30 mins	Inhibition of mTORC1 in HEK293 cells using GST-tagged S6K1 or Akt1 as substrate after 30 mins by immunoblotting assay, IC50=0.01μM	29211480
HEK293	Function assay		30 mins	Inhibition of mTORC2 in HEK293 cells using GST-tagged S6K1 or Akt1 as substrate after 30 mins by immunoblotting assay, IC50=0.01μM	29211480
NUGC4	Growth Inhibition Assay			IC50=2.93 ± 0.31 μM	24597478
MKN45	Growth Inhibition Assay			IC50=0.82 ± 0.01 μM	24597478
HGC27	Growth Inhibition Assay			IC50=15.0 ± 4.82 μM	24597478
AGS	Growth Inhibition Assay			IC50=15.0 ± 2.91 μM	24597478
HEK293	Function assay			Inhibition of recombinant FLAG-tagged mTOR (1362 to 2549) (unknown origin) expressed in HEK293 cells, IC50=0.0025μM	23375793
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
BT-37	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
NB-EBc1	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
OHS-50	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
MG 63 (6-TG R)	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells	29435139
Rh41	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for A673 cells)	29435139
BT-12	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-12 cells	29435139
DAOY	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for DAOY cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SJ-GBM2 cells	29435139
U-2 OS	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for U-2 OS cells	29435139
Rh41	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh41 cells	29435139
RD	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for RD cells	29435139
Rh18	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh18 cells	29435139
Rh30	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh30 cells	29435139
SK-N-SH	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-SH cells	29435139
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生物活性

製品説明

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
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]
Kinase Assay	mTOR complexes kinase assays
Kinase Assay	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.
細胞実験	細胞株	Wild-type and mLST8 deficient MEFs
	濃度	Dissolved in DMSO, final concentration ~3 μM
	反応時間	24, 48, and 72 hours
	実験の流れ	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.
実験結果図	Methods	Biomarkers	結果図	PMID
実験結果図	Western blot	p-S6K / S6K / p-4E-BP1 / E7 / E6 / p53 p-mTOR		28115701

In Vivo
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].
動物実験	動物モデル	Nu/Nu nude mice
	投与量	8 mg/kg
	投与経路	i.p.

参考
[1] García-Martínez JM, et al. Biochem J, 2009, 421(1), 29 [2] Zhang H, et al. PLoS One. 2013, 8(1):e54918.

参考

化学情報

分子量	465.54	化学式	C₂₅H₃₁N₅O₄
CAS No.	938440-64-3	SDF	Download KU-0063794 SDFをダウンロードする
Smiles	CC1CN(CC(O1)C)C2=NC3=C(C=CC(=N3)C4=CC(=C(C=C4)OC)CO)C(=N2)N5CCOCC5
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1年-80°Cin solvent
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1个月-20°Cin solvent

In vitro
Batch:

DMSO : 16 mg/mL ( (34.36 mM)；吸湿したDMSOは溶解度を減少させます。新しいDMSOをご使用ください。)

Ethanol : 5 mg/mL

Water : Insoluble

モル濃度計算器

in vivo
Batch:

Add solvents to the product individually and in order.

投与溶液組成計算機

実験計算

モル濃度計算器

質量	濃度	体積	分子量
=	×	×

希釈計算器分子量計算器

投与溶液組成計算機(クリア溶液)

ステップ１：実験データを入力してください。（実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します）

投与量 mg/kg 動物平均体重 g 投与体積（動物毎）μL 動物数匹

ステップ２：投与溶媒の組成を入力してください。（ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください）

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

計算結果：

投与溶媒濃度： mg/ml;

DMSOストック溶液調製方法： mg 試薬を μL DMSOに溶解する（濃度 mg/mL, 注：濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法：Take μL DMSOストック溶液に μL PEG300，を加え、完全溶解後μL Tween 80，を加えて完全溶解させた後 μL ddH₂O，を加え完全に溶解させます。

投与溶媒調製方法：μL DMSOストック溶液に μL Corn oil，を加え、完全溶解。

注意：１.ストック溶液に沈殿、混濁などがないことをご確認ください；
２.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):