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KU-0063794 mTOR inhibitor

Name: KU-0063794
Brand: Selleck Chemicals
SKU: S1226
Rating: 5 (77 reviews)

N. Cat.: S1226

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.

KU-0063794 mTOR inhibitor Chemical Structure

Struttura chimica

Peso molecolare: 465.54

Vai a

Controllo Qualità (Quality Control)

Lotto: Purezza: 99.91%

99.91

Target correlati	PI3K Akt GSK-3 ATM/ATR DNA-PK AMPK PDPK1 PTEN PP2A PDK
Altro mTOR Inibitori	Torin 1 Torin 2 AZD8055 Ridaforolimus (Deforolimus, MK-8669) Sapanisertib (MLN0128, INK-128) Torkinib (PP242) MHY1485 Vistusertib (AZD2014) OSI-027 3BDO

Coltura cellulare, trattamento e concentrazione di lavoro
(Cell Culture, Treatment & Working Concentration)

Linee cellulari	Tipo di saggio	Concentrazione	Tempo di incubazione	Formulazione	Descrizione dell'attività	PMID
HepG2	Cell Viability Assay	0.1–50 μM	72 h		decreases cell viability 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	Apoptosis Assay	0.1–50 μM	48 h		induces apoptosis in a dose dependent manner	26278819
HepG2	Function Assay	5/10 μM	24 h		dramatically inhibits phosphorylation of AKT at Ser-473	26278819
HepG2	Function Assay	5/10 μM	24 h		downregulates the levels HIF1α and cyclin D1	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
HepG3	Function Assay	0.1–50 μM	48 h		induces cell autophagy in a dose dependent manner	26278819
AGS	Growth Inhibition Assay				IC50=15.0 ± 2.91 μM	24597478
HGC27	Growth Inhibition Assay				IC50=15.0 ± 4.82 μM	24597478
MKN45	Growth Inhibition Assay				IC50=0.82 ± 0.01 μM	24597478
NUGC4	Growth Inhibition Assay				IC50=2.93 ± 0.31 μM	24597478
PC9	Growth Inhibition Assay		72 h		IC50=10.15±0.62 nM	23874880
PC9GR	Growth Inhibition Assay		72 h		IC50=6.21±1.30 nM	23874880
H1650	Growth Inhibition Assay		72 h		IC50=7.61±0.62 nM	23874880
H1975	Growth Inhibition Assay		72 h		IC50=11.15±0.93 nM	23874880
PC9	Function Assay	10.15 nM	72 h		inhibits mTOR phosphorylation status	23874880
PC9GR	Function Assay	6.21 nM	72 h		inhibits mTOR phosphorylation status	23874880
H1650	Function Assay	7.61 nM	72 h		inhibits mTOR phosphorylation status	23874880
H1975	Function Assay	11.15 nM	72 h		inhibits mTOR phosphorylation status	23874880
PC9	Function Assay	10.15 nM	72 h		inhibits phosphorylation of p70S6K	23874880
PC9GR	Function Assay	6.21 nM	72 h		inhibits phosphorylation of p70S6K	23874880
H1650	Function Assay	7.61 nM	72 h		inhibits phosphorylation of p70S6K	23874880
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	DMSO	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	DMSO	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	DMSO	suppresses the cell viability in both time and dose dependent manner	23349989
786-O	Cell Viability Assay	300-4000 nM	24-96 h	DMSO	suppresses the cell viability in both time and dose dependent manner	23349989
Caki-1	Function Assay	2 µM	72 h	DMSO	induces G1 cell cycle arrest and autophagy	23349989
786-O	Function Assay	2 µM	72 h	DMSO	induces G1 cell cycle arrest and autophagy	23349989
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
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
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
HEK293	Function assay				Inhibition of recombinant FLAG-tagged mTOR (1362 to 2549) (unknown origin) expressed in HEK293 cells, IC50=0.0025μM	23375793
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
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
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
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
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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Informazioni chimiche, conservazione e stabilità (Chemical Information, Storage & Stability)

Peso molecolare	465.54	Formula	C₂₅H₃₁N₅O₄	Conservazione (Dalla data di ricezione)	3 anni-20°Cpolvere
N. CAS	938440-64-3	Scarica SDF		Conservazione delle soluzioni stock	1 anno-80°Cin solvente 1 mese-20°Cin solvente
Sinonimi	N/A			Smiles	CC1CN(CC(O1)C)C2=NC3=C(C=CC(=N3)C4=CC(=C(C=C4)OC)CO)C(=N2)N5CCOCC5

Solubilità (Solubility)

In vitro
Lotto:

DMSO : 16 mg/mL (34.36 mM)
(Il DMSO contaminato da umidità può ridurre la solubilità. Utilizzare DMSO fresco e anidro.)

Ethanol : 5 mg/mL

Water : Insoluble

Calcolatore di Molarità

Massa	Concentrazione	Volume	Peso molecolare
=	×	×

Calcolatore di Diluizione Calcolatore del Peso Molecolare

In vivo Lotto:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	30%PEG400 1 0.5%Tween80 2 5%propylene glycol Validato dai laboratori Selleck. Se necessiti di modifiche a questa formulazione, contatta il nostro team di vendita per test personalizzati.	30.000mg/ml (64.44mM)	Taking the 1 mL working solution as an example, add 300 μL of 100 mg/ml clarified PEG400 stock solution to 5 μL of Tween80, mix evenly to clarify it; add 50 μL Propylene glycol to the above system, mix evenly to clarify it; then continue to add 645 μL ddH2O to adjust the volume. to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

Calcolatore di formulazione in vivo (Soluzione chiara)

Passo 1: Inserire le informazioni di seguito (Consigliato: Un animale aggiuntivo per tenere conto della perdita durante l'esperimento)

Dosaggio: mg/kg Peso medio degli animali: g Volume di dosaggio per animale:μL Numero di animali:

Passo 2: Inserire la formulazione in vivo (Questo è solo il calcolatore, non la formulazione. Contattateci prima se non c'è una formulazione in vivo nella sezione Solubilità.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Risultati del calcolo:

Concentrazione di lavoro: mg/ml;

Metodo per preparare il liquido master di DMSO: mg farmaco predissolto in μL DMSO ( Concentrazione del liquido master mg/mL, Vi preghiamo di contattarci prima se la concentrazione supera la solubilità del DMSO del lotto del farmaco. )

Metodo per preparare la formulazione in vivo: Prendere μL DMSO liquido master, quindi aggiungereμL PEG300, mescolare e chiarire, quindi aggiungereμL Tween 80, mescolare e chiarire, quindi aggiungere μL ddH₂O, mescolare e chiarire.

Metodo per preparare la formulazione in vivo: Prendere μL DMSO liquido master, quindi aggiungere μL Olio di mais, mescolare e chiarire.

Nota: 1. Si prega di assicurarsi che il liquido sia limpido prima di aggiungere il solvente successivo.
2. Assicurarsi di aggiungere il/i solvente/i in ordine. È necessario assicurarsi che la soluzione ottenuta, nell'aggiunta precedente, sia una soluzione limpida prima di procedere all'aggiunta del solvente successivo. Metodi fisici come il vortex, gli ultrasuoni o il bagno d'acqua calda possono essere utilizzati per facilitare la dissoluzione.

Meccanismo d'azione (Mechanism of Action)

Targets/IC₅₀/Ki	mTORC1 (Cell-free assay) ~10 nM mTORC2 (Cell-free assay) ~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, this compound 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 this compound is needed to inhibit the S6K1 activity by ~90%. This chemical at 100-300 nM also completely inhibits the amino-acid-induced phosphorylation of S6K1 and S6 protein. Similar to S6K1, it 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, this compound 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. This compound 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 it dose not inhibit phorbol ester induced ERK or RSK phosphorylation and RSK activation. Compared with rapamycin, this chemical exhibits more significant potency to induce the complete dephosphorylation of 4E-BP1 at Thr³⁷, Thr⁴⁶ and Ser⁶⁵. It inhibits cell growth of both wild-type and mLST8-deficient MEFs and induces a G1 cell cycle arrest, more significantly than rapamycin.
Saggio chinasico	Saggi chinasici dei complessi di mTOR
Saggio chinasico	Le cellule HEK-293 vengono lisate fresche in tampone di lisi Hepes. Il lisato (1-4 mg) viene pre-chiarificato incubando con 5-20 μL di Proteina G-Sepharose coniugata a IgG pre-immune. Gli estratti del lisato vengono quindi incubati con 5-20 μL di Proteina G-Sepharose coniugata a 5-20 μg di anticorpo anti-Rictor o anti-Raptor, o IgG pre-immune. Tutti gli anticorpi sono coniugati covalentemente a Proteina G-Sepharose. Le immunoprecipitazioni vengono eseguite per 1 ora a 4 °C su una piattaforma vibrante. Gli immunoprecipitati vengono lavati quattro volte con tampone di lisi Hepes, seguiti da due lavaggi con tampone chinasico Hepes. Per gli immunoprecipitati di Raptor utilizzati per fosforilare S6K1, per i due passaggi di lavaggio iniziali il tampone include 0.5 M NaCl per garantire un'attività chinasica ottimale. GST-Akt1 è isolata da cellule HEK-293 private di siero incubate con PI-103 (1 μM per 1 ora). GST-S6K1 è purificata da cellule HEK-293 private di siero incubate con rapamicina (0.1 μM per 1 ora). Le reazioni di mTOR vengono avviate aggiungendo 0.1 mM ATP e 10 mM MgCl₂ in presenza di varie concentrazioni di KU-0063794 e GST-Akt1 (0.5 μg) o GST-S6K1 (0.5 μg). Le reazioni vengono eseguite per 30 minuti a 30 °C su una piattaforma vibrante e interrotte con l'aggiunta di tampone SDS per campioni. Le miscele di reazione vengono quindi filtrate attraverso un filtro Spin-X con pori da 0.22 μm e i campioni vengono sottoposti a elettroforesi e analisi di immunoblotting con gli anticorpi indicati.
In vivo	Ku0063794 inhibits tumor growth and mTOR signaling in a preclinical renal cell carcinoma model. However, this compound was not more effective in the animal study. A possible explanation for lack of greater activity in vivo for this chemical.
Riferimenti	[1] https://pubmed.ncbi.nlm.nih.gov/19402821/ [2] https://pubmed.ncbi.nlm.nih.gov/23349989/

Applicazioni (Applications)

Metodi	Biomarcatori	Immagini	PMID
Western blot	p-mTOR p-S6K / S6K / p-4E-BP1 / E7 / E6 / p53		24262658

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):