solo para uso en investigación

KU-0063794 mTOR Inhibidor

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

Cat. No.: S1226

KU-0063794 es un inhibidor dual de mTOR potente y altamente específico de mTORC1 y mTORC2 con una IC50 de ~10 nM en ensayos sin células; sin efecto sobre PI3Ks.

KU-0063794 mTOR Inhibidor Chemical Structure

Estructura química

Peso molecular: 465.54

Saltar a

Control de calidad (Quality Control)

Lote: Pureza: 99.91%

99.91

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

Cultivo celular, tratamiento y concentración de trabajo
(Cell Culture, Treatment & Working Concentration)

Líneas celulares	Tipo de ensayo	Concentración	Tiempo de incubación	Formulación	Descripción de la actividad	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
Haga clic para ver más datos experimentales de líneas celulares

Información química, almacenamiento y estabilidad (Chemical Information, Storage & Stability)

Peso molecular	465.54	Fórmula	C₂₅H₃₁N₅O₄	Almacenamiento (Desde la fecha de recepción)	3 años-20°Cpolvo
Nº CAS	938440-64-3	Descargar SDF		Almacenamiento de soluciones madre	1 año-80°Cen disolvente 1 mes-20°Cen disolvente
Sinónimos	N/A			Smiles	CC1CN(CC(O1)C)C2=NC3=C(C=CC(=N3)C4=CC(=C(C=C4)OC)CO)C(=N2)N5CCOCC5

Solubilidad (Solubility)

In vitro
Lote:

DMSO : 16 mg/mL (34.36 mM)
(El DMSO contaminado con humedad puede reducir la solubilidad. Usar DMSO fresco y anhidro.)

Ethanol : 5 mg/mL

Water : Insoluble

Calculadora de Molaridad

Masa	Concentración	Volumen	Peso molecular
=	×	×

Calculadora de Dilución Calculadora de Peso Molecular

In vivo Lote:
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 Validado por los laboratorios Selleck. Si necesita ajustes en esta formulación, contacte con nuestro equipo de ventas para pruebas personalizadas.	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.

Calculadora de formulación in vivo (Solución clara)

Paso 1: Introduzca la información a continuación (Recomendado: Un animal adicional para tener en cuenta la pérdida durante el experimento)

Dosis: mg/kg Peso promedio de los animales: g Volumen de dosificación por animal:μL Número de animales:

Paso 2: Introduzca la formulación in vivo (Esto es solo la calculadora, no la formulación. Por favor, contáctenos primero si no hay una formulación in vivo en la sección de Solubilidad.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Resultados del cálculo:

Concentración de trabajo: mg/ml;

Método para preparar el líquido maestro de DMSO: mg fármaco predissuelto en μL DMSO ( Concentración del líquido maestro mg/mL, Por favor, contáctenos primero si la concentración excede la solubilidad del DMSO del lote del fármaco. )

Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadirμL PEG300, mezclar y clarificar, luego añadirμL Tween 80, mezclar y clarificar, luego añadir μL ddH₂O, mezclar y clarificar.

Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadir μL Aceite de maíz, mezclar y clarificar.

Nota: 1. Por favor, asegúrese de que el líquido esté claro antes de añadir el siguiente disolvente.
2. Asegúrese de añadir el (los) disolvente(s) en orden. Debe asegurarse de que la solución obtenida, en la adición anterior, sea una solución clara antes de proceder a añadir el siguiente disolvente. Se pueden utilizar métodos físicos como el vórtice, el ultrasonido o el baño de agua caliente para ayudar a la disolución.

Mecanismo de acción (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.
Ensayo de quinasa	Ensayos de quinasas de complejos mTOR
Ensayo de quinasa	Las células HEK-293 se lisan frescas en tampón de lisis Hepes. El lisado (1-4 mg) se pre-clarifica incubando con 5-20 μL de Proteína G-Sepharose conjugada a IgG preinmune. Los extractos de lisado se incuban luego con 5-20 μL de Proteína G-Sepharose conjugada a 5-20 μg de anticuerpo anti-Rictor o anti-Raptor, o IgG preinmune. Todos los anticuerpos están conjugados covalentemente a Proteína G-Sepharose. Las inmunoprecipitaciones se realizan durante 1 hora a 4 °C en una plataforma vibratoria. Los inmunoprecipitados se lavan cuatro veces con tampón de lisis Hepes, seguido de dos lavados con tampón de quinasa Hepes. Para los inmunoprecipitados de Raptor utilizados para fosforilar S6K1, para los dos pasos de lavado iniciales el tampón incluye 0.5 M de NaCl para asegurar una actividad quinasa óptima. La GST-Akt1 se aísla de células HEK-293 privadas de suero incubadas con PI-103 (1 μM durante 1 hora). La GST-S6K1 se purifica de células HEK-293 privadas de suero incubadas con rapamicina (0.1 μM durante 1 hora). Las reacciones de mTOR se inician añadiendo 0.1 mM de ATP y 10 mM de MgCl₂ en presencia de varias concentraciones de KU-0063794 y GST-Akt1 (0.5 μg) o GST-S6K1 (0.5 μg). Las reacciones se realizan durante 30 minutos a 30 °C en una plataforma vibratoria y se detienen añadiendo tampón de muestra SDS. Las mezclas de reacción se filtran a través de un filtro Spin-X de 0.22 μm de tamaño de poro y las muestras se someten a electroforesis y análisis de inmunotransferencia con los anticuerpos indicados.
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.
Referencias	[1] https://pubmed.ncbi.nlm.nih.gov/19402821/ [2] https://pubmed.ncbi.nlm.nih.gov/23349989/

Aplicaciones (Applications)

Métodos	Biomarcadores	Imágenes	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):