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

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Kwaliteitscontrole (Quality Control)

Batch: Zuiverheid: 99.98%

99.98

Gerelateerde doelwitten	Akt mTOR GSK-3 ATM/ATR DNA-PK AMPK PDPK1 PTEN PI4K PP2A
Overig PI3K Inhibitoren	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)

Celkweek, behandeling & werkzame concentratie
(Cell Culture, Treatment & Working Concentration)

Cellijnen	Assaytype	Concentratie	Incubatietijd	Activiteitsbeschrijving
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
Klik om meer experimentele gegevens over cellijnen te bekijken

Chemische informatie, opslag en stabiliteit (Chemical Information, Storage & Stability)

Molecuulgewicht	417.41	Formule	C₁₉H₂₁F₂N₇O₂	Opslag (vanaf de datum van ontvangst)	3 jaar-20°Cpoeder
CAS-nr.	475110-96-4	SDF downloaden		Opslag van stamoplossingen	1 jaar-80°Cin oplosmiddel 1 maand-20°Cin oplosmiddel
Synoniemen	N/A			Smiles	C1COCCN1C2=NC(=NC(=N2)N3C4=CC=CC=C4N=C3C(F)F)N5CCOCC5

Oplosbaarheid (Solubility)

In vitro
Batch:

DMSO : 15 mg/mL (35.93 mM)
(Met vocht verontreinigd DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molariteitscalculator

Massa	Concentratie	Volume	Molecuulgewicht
=	×	×

Verdunningscalculator Molecuulgewichtcalculator

In vivo Batch:
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 Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	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.

In vivo formulatiecalculator (heldere oplossing)

Stap 1: Voer onderstaande informatie in (Aanbevolen: een extra dier om rekening te houden met verlies tijdens het experiment)

Dosering: mg/kg Gemiddeld gewicht van dieren: g Doseervolume per dier:μL Aantal dieren:

Stap 2: Voer de in vivo formulering in (Dit is alleen de calculator, geen formulering. Neem eerst contact met ons op als er geen in vivo formulering is in de sectie oplosbaarheid.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Berekeningsresultaten:

Werkconcentratie: mg/ml;

Methode voor het bereiden van DMSO-moedervloeistof: mg geneesmiddel vooropgelost in μL DMSO ( Concentratie moedervloeistof mg/mL, Neem eerst contact met ons op als de concentratie de DMSO-oplosbaarheid van de batch van het geneesmiddel overschrijdt. )

Methode voor het bereiden van in vivo formulering: Neem μL DMSO moedervloeistof, voeg daarna toeμL PEG300, mengen en verhelderen, daarna toevoegenμL Tween 80, mengen en verhelderen, daarna toevoegen μL ddH₂O, mengen en verhelderen.

Methode voor het bereiden van in vivo formulering: Neem μL DMSO moedervloeistof, voeg daarna toe μL Maïsolie, mengen en verhelderen.

Opmerking: 1. Zorg ervoor dat de vloeistof helder is voordat u het volgende oplosmiddel toevoegt.
2. Zorg ervoor dat u het/de oplosmiddel(en) in de juiste volgorde toevoegt. U moet ervoor zorgen dat de verkregen oplossing, bij de vorige toevoeging, een heldere oplossing is voordat u verdergaat met het toevoegen van het volgende oplosmiddel. Fysieke methoden zoals vortexen, ultrasoon of een warmwaterbad kunnen worden gebruikt om het oplossen te bevorderen.

Werkingsmechanisme (Mechanism of Action)

Kenmerken	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).
Kinase Assay	Remming van PI3K-activiteit
Kinase Assay	A549-cellen worden gelyseerd in een buffer die 20 mM Tris-HCl (pH 7,5), 150 mM NaCl, 5 mM EDTA en 1% Igepal CA-630 bevat, de lysaten worden gedurende 10 minuten gecentrifugeerd bij 20.000 g en 4 °C, en de supernatanten worden gebruikt als cellysaat (eiwit = 2-4 mg/mL). Om PI3K te immunoprecipiteren, worden 200 μL cellysaat geïncubeerd met anti-p85 polyklonaal antilichaam en proteïne G-agarose (5 μL). PI3Kα, PI3Kβ en PI3Kδ kunnen worden geïmmunoprecipiteerd door het anti-p85 polyklonale antilichaam. Agarosekralen die immunoprecipitaten bevatten, worden tweemaal gewassen met buffer A (20 mM Tris-HCl bij pH 7,5, 150 mM NaCl, 5 mM EDTA en 1% Igepal CA-630), eenmaal met buffer B (500 mM LiCl en 100 mM Tris-HCl bij pH 7,5), eenmaal met gedestilleerd water en eenmaal met buffer C (100 mM NaCl en 20 mM Tris-HCl bij pH 7,5). Immunoprecipitaten worden gesuspendeerd in 20 μL buffer C die fosfatidylinositol van 200 μg/mL bevat. Het mengsel wordt 5 minuten voorgeïncubeerd met toenemende concentraties van deze verbinding bij 25 °C. [γ-³²P]ATP (2 μCi per assaymengsel; eindconcentratie, 20 μM) en MgCl₂ (eindconcentratie, 20 mM) worden toegevoegd om de reactie te starten. Het reactiemengsel wordt gedurende 20 minuten geïncubeerd bij 25 °C. Gefosforyleerde producten van fosfatidylinositol worden gescheiden door dunne-laagchromatografie en gevisualiseerd door autoradiografie. Het fosfatidylinositol-3-fosfaatgebied wordt van de plaat geschraapt en de radioactiviteit wordt ook gemeten met vloeistofscintillatiespectroscopie. Het remmingsniveau voor deze chemische stof wordt bepaald als het percentage ³²P-tellingen per minuut verkregen zonder deze verbinding.
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.
Referenties	[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/

Toepassingen (Applications)

Methoden	Biomarkers	Afbeeldingen	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

Informatie over klinische proeven (Clinical Trial Information)

(gegevens van https://clinicaltrials.gov, bijgewerkt op 2017-07-06)

NCT-nummer	Werving	Aandoeningen	Sponsor/medewerkers	Startdatum	Fasen
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 Remmer

Molecuulgewicht: 417.41

Kwaliteitscontrole (Quality Control)

Celkweek, behandeling & werkzame concentratie
(Cell Culture, Treatment & Working Concentration)

Chemische informatie, opslag en stabiliteit (Chemical Information, Storage & Stability)

Oplosbaarheid (Solubility)

Molariteitscalculator

In vivo formulatiecalculator (heldere oplossing)

Werkingsmechanisme (Mechanism of Action)

Toepassingen (Applications)

Informatie over klinische proeven (Clinical Trial Information)

ZSTK474 PI3K Remmer

Molecuulgewicht: 417.41

Kwaliteitscontrole (Quality Control)

Celkweek, behandeling & werkzame concentratie (Cell Culture, Treatment & Working Concentration)

Chemische informatie, opslag en stabiliteit (Chemical Information, Storage & Stability)

Oplosbaarheid (Solubility)

Molariteitscalculator

In vivo formulatiecalculator (heldere oplossing)

Werkingsmechanisme (Mechanism of Action)

Toepassingen (Applications)

Informatie over klinische proeven (Clinical Trial Information)

Celkweek, behandeling & werkzame concentratie
(Cell Culture, Treatment & Working Concentration)