Name: ZSTK474
Brand: Selleck Chemicals
SKU: S1072
Rating: 5 (85 reviews)

Saltar a

Control de calidad (Quality Control)

Lote: Pureza: 99.98%

99.98

Dianas relacionadas	Akt mTOR GSK-3 ATM/ATR DNA-PK AMPK PDPK1 PTEN PI4K PP2A
Otros PI3K Inhibidores	GDC-0077 (Inavolisib) SAR405 Quercetin (Sophoretin) LY294002 XL147 analogue Tersolisib (STX-478) Buparlisib (BKM120) 740 Y-P (PDGFR 740Y-P) GO-203 TFA Eganelisib (IPI-549)

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

Líneas celulares	Tipo de ensayo	Concentración	Tiempo de incubación	Descripción de la actividad
Sf21 insect cells	Function assay		1 h	Inhibition of bovine recombinant PI3K p110delta expressed in Sf21 insect cells using phosphatidylinositol as substrate after 1 hr by phosphoimaging, IC50=0.7 nM
human HCT116 cells	Function assay		15 mins	Inhibition of PIK3CA H1047R mutant-mediated cell signaling in human HCT116 cells expressing PTEN assessed as inhibition of insulin-induced pAkt/PKB phosphorylation at Thr308 treated for 15 mins before insulin challenge measured after 5 mins by immunoblotting, IC50=78 nM
human LNCAP cells	Proliferation assay		3 days	Antiproliferative activity against human LNCAP cells after 3 days by MTS assay, IC50=0.21 μM
human NZB5 cells	Proliferation assay		5 days	Antiproliferative activity against human NZB5 cells expressing wild type p110alpha assessed as incorporation of [3H]thymidine after 5 days, IC50=0.22 μM
human NZOV9 cells	Proliferation assay		5 days	Antiproliferative activity against human NZOV9 cells expressing p110alpha kinase Y1021C mutant assessed as incorporation of [3H]thymidine after 5 days, IC50=0.29 μM
human MDA-MB-468 cells	Cytotoxic assay		48 h	Cytotoxicity against PTEN-deficient human MDA-MB-468 cells assessed as inhibition of cell growth after 48 hrs by Cell Titer 96 assay
human A549 cells	Function assay	10 μM	1 h	Inhibition of PI3K in human A549 cells assessed as reduction in pAkt level at 10 uM after 1 hr by Western blotting analysis
human DMS114 cells	Growth inhibition assay			Growth inhibition of human DMS114 cells
human MKN74 cells	Growth inhibition assay			Growth inhibition of human MKN74 cells
human SNB78 cells	Growth inhibition assay			Growth inhibition of human SNB78 cells
human St-4 cells	Growth inhibition assay			Growth inhibition of human St-4 cells
human DU145 cells	Growth inhibition assay			Growth inhibition of human DU145 cells
human LOXIMVI cells	Growth inhibition assay			Growth inhibition of human LOXIMVI cells
human PC3 cells	Growth inhibition assay			Growth inhibition of human PC3 cells
human LOXIMVI cells	Growth inhibition assay			Growth inhibition of human LOXIMVI cells
human OVCAR3 cells	Growth inhibition assay			Growth inhibition of human OVCAR3 cells
human SKOV3 cells	Growth inhibition assay			Growth inhibition of human SKOV3 cells
human KM12 cells	Growth inhibition assay			Growth inhibition of human KM12 cells
human HT-29 cells	Growth inhibition assay			Growth inhibition of human HT-29 cells
human HCT15 cells	Growth inhibition assay			Growth inhibition of human HCT15 cells
human NCI-H226 cells	Growth inhibition assay			Growth inhibition of human NCI-H226 cells
human NCI-H522 cells	Growth inhibition assay			Growth inhibition of human NCI-H522 cells
human A549 cells	Growth inhibition assay			Growth inhibition of human A549 cells
human HCC2998 cells	Growth inhibition assay			Growth inhibition of human HCC2998 cells
human SNB75 cells	Growth inhibition assay			Growth inhibition of human SNB75 cells
human OVCAR4 cells	Growth inhibition assay			Growth inhibition of human OVCAR4 cells
human OVCAR5 cells	Growth inhibition assay			Growth inhibition of human OVCAR5 cells
human OVCAR8 cells	Growth inhibition assay			Growth inhibition of human OVCAR8 cells
human SKOV3 cells	Growth inhibition assay			Growth inhibition of human SKOV3 cells
human ACHN cells	Growth inhibition assay			Growth inhibition of human ACHN cells
Haga clic para ver más datos experimentales de líneas celulares

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

Peso molecular	417.41	Fórmula	C₁₉H₂₁F₂N₇O₂	Almacenamiento (Desde la fecha de recepción)	3 años-20°Cpolvo
Nº CAS	475110-96-4	Descargar SDF		Almacenamiento de soluciones madre	1 año-80°Cen disolvente 1 mes-20°Cen disolvente
Sinónimos	N/A			Smiles	C1COCCN1C2=NC(=NC(=N2)N3C4=CC=CC=C4N=C3C(F)F)N5CCOCC5

Solubilidad (Solubility)

In vitro
Lote:

DMSO : 15 mg/mL (35.93 mM)
(El DMSO contaminado con humedad puede reducir la solubilidad. Usar DMSO fresco y anhidro.)

Water : Insoluble

Ethanol : 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	0.5% hydroxyethyl cellulose Validado por los laboratorios Selleck. Si necesita ajustes en esta formulación, contacte con nuestro equipo de ventas para pruebas personalizadas.	10.000mg/ml (23.96mM)	Taking the 1 mL working solution as an example, take 10 mg of this product, add it to 1 ml of 0.5% hydroxyethyl cellulose clear solution, and mix evenly to form a uniform suspension. 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)

Características	First orally administered PI3K inhibitor used in vivo.
Targets/IC₅₀/Ki	PI3Kδ (Cell-free assay) 4.6 nM PI3Kα (Cell-free assay) 16 nM PI3K (Cell-free assay) 37 nM PI3Kβ (Cell-free assay) 44 nM PI3Kγ (Cell-free assay) 49 nM
In vitro	ZSTK474 at 1 μM potently reduces PI3K activity to 4.7% of the control level, whereas LY2194002 only reduces the activity to 44.6% of the control. This compound inhibits the activities of recombinant p110β, -γ, and -δ with IC50 of 17 nM, 53 nM, and 6 nM, respectively. It shows potent antiproliferative activity against a panel of 39 human cancer cell lines with mean GI50 of 0.32 μM, more effectively than that of LY294002 or wortmannin with mean GI50 of 7.4 μM or 10 μM, respectively. This chemical treatment at 1 μM blocks membrane ruffling and generation of PIP3 induced by platelet-derived growth factor in murine embryonic fibroblasts (MEFs). It at 10 μM induces apoptosis in OVCAR3 cells, and induces complete G1-phase arrest but not apoptosis in A549 cells. This compound treatment at 0.5 μM significantly decreases the level of phosphorylated Akt and GSK-3β, as well as the cyclin D1 protein expression. It also inhibits the phosphorylation of other downstream signaling components that are involved in regulating cell proliferation including FKHRL1, FKHR, TSC-2, mTOR, and p70S6K in a dose-dependent manner. This chemical does not inhibit mTOR at 0.1 μM, and even at a concentration of 100 μM, it inhibits mTOR activity less than 40%. It blocks VEGF-induced cell migration and the tube formation in human umbilical vein endothelial cells (HUVECs), and inhibits the expression of HIF-1α and secretion of VEGF in RXF-631L cells, exhibiting potent in vitro antiangiogenic activity. This compound treatment inhibits the production of IFNγ and IL-17 in concanavalin A-activated T cells, and inhibits the proliferation and PGE(2) production by fibroblast-like synovial cells (FLS).
Ensayo de quinasa	Inhibición de la actividad de PI3K
Ensayo de quinasa	Las células A549 se lisan en un tampón que contiene 20 mM Tris-HCl (pH 7,5), 150 mM NaCl, 5 mM EDTA y 1% de Igepal CA-630, los lisados se centrifugan a 20.000 g y 4 °C durante 10 minutos, y los sobrenadantes se utilizan como lisado celular (proteína = 2-4 mg/mL). Para inmunoprecipitar PI3K, se incuban 200 μL de lisado celular con anticuerpo policlonal anti-p85 y agarosa de proteína G (5 μL). Las PI3Kα, PI3Kβ y PI3Kδ pueden ser inmunoprecipitadas por el anticuerpo policlonal anti-p85. Las perlas de agarosa que contienen los inmunoprecipitados se lavan dos veces con el tampón A (20 mM Tris-HCl a pH 7,5, 150 mM NaCl, 5 mM EDTA y 1% de Igepal CA-630), una vez con el tampón B (500 mM LiCl y 100 mM Tris-HCl a pH 7,5), una vez con agua destilada y una vez con el tampón C (100 mM NaCl y 20 mM Tris-HCl a pH 7,5). Los inmunoprecipitados se suspenden en 20 μL del tampón C que contiene fosfatidilinositol a 200 μg/mL. La mezcla se preincuba con concentraciones crecientes de este compuesto a 25 °C durante 5 minutos. Se añaden [γ-³²P]ATP (2 μCi por mezcla de ensayo; concentración final, 20 μM) y MgCl₂ (concentración final, 20 mM) para iniciar la reacción. La mezcla de reacción se incuba a 25 °C durante 20 minutos. Los productos fosforilados del fosfatidilinositol se separan por cromatografía en capa fina y se visualizan por autorradiografía. La región del fosfatidilinositol-3-fosfato se raspa de la placa, y la radioactividad también se mide con espectroscopia de centelleo líquido. El nivel de inhibición para este químico se determina como el porcentaje de recuentos de ³²P por minuto obtenidos sin este compuesto.
In vivo	Oral administration of ZSTK474 inhibits the growth of subcutaneously implanted mouse B16F10 melanoma tumors in a dose-dependent manner, producing tumor regression of 28.5%, 7.1%, or 4.9% on day 14 at 100, 200, or 400 mg/kg, respectively, which is superior to that of the four major anticancer drugs at their respective maximum tolerable doses with tumor regression of 96%, 35.7%, 24%, or 68.3%, respectively. This compound treatment at 400 mg/kg completely inhibits the growth of A549, PC-3, and WiDr xenografts in mice, and induces the regression of A549 xenograft tumors. It significantly inhibits tumor growth in the RXF-631L xenograft model, correlated with a significantly reduced number of microvessels in the treated mice. Oral administration of this chemical ameliorates the progression of adjuvant-induced arthritis (AIA) in rats.
Referencias	[1] https://pubmed.ncbi.nlm.nih.gov/16622124/ [2] https://pubmed.ncbi.nlm.nih.gov/17711503/ [3] https://pubmed.ncbi.nlm.nih.gov/19144509/ [4] https://pubmed.ncbi.nlm.nih.gov/19372588/ [5] https://pubmed.ncbi.nlm.nih.gov/22039466/ [6] https://pubmed.ncbi.nlm.nih.gov/22349137/

Aplicaciones (Applications)

Métodos	Biomarcadores	Imágenes	PMID
Western blot	p-PDK1 / p-GSK3β / p-AKT / AKT P-gp / MRP1 p-Rb / p27 / Cyclin D1 HIF-1α / HIF-1β		28388564
Growth inhibition assay	Cell viability		28388564

Información del ensayo clínico (Clinical Trial Information)

(datos de https://clinicaltrials.gov, actualizado el 2017-07-06)

Número NCT	Reclutamiento	Condiciones	Patrocinador/Colaboradores	Fecha de inicio	Fases
NCT01682473	COMPLETED	Neoplasms	Zenyaku Kogyo Co., Ltd.	2012-09-20	PHASE1
NCT01280487	COMPLETED	Neoplasms	Zenyaku Kogyo Co., Ltd.	2011-01	PHASE1

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):

ZSTK474 PI3K Inhibidor

Peso molecular: 417.41

Control de calidad (Quality Control)

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

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

Solubilidad (Solubility)

Calculadora de Molaridad

Calculadora de formulación in vivo (Solución clara)

Mecanismo de acción (Mechanism of Action)

Aplicaciones (Applications)

Información del ensayo clínico (Clinical Trial Information)

ZSTK474 PI3K Inhibidor

Peso molecular: 417.41

Control de calidad (Quality Control)

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

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

Solubilidad (Solubility)

Calculadora de Molaridad

Calculadora de formulación in vivo (Solución clara)

Mecanismo de acción (Mechanism of Action)

Aplicaciones (Applications)

Información del ensayo clínico (Clinical Trial Information)

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