ZSTK474

ZSTK474 inhibits class I PI3K isoforms with IC50 of 37 nM in a cell-free assay, mostly PI3Kδ. Phase1/2.

CAS No. 475110-96-4

サイズ	価格(税別)	在庫状況
10mM (1mL in DMSO)	JPY 16900	国内在庫あり
10mg	JPY 16200	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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ZSTK474関連製品

関連ターゲット	p110α p110β p110δ p110γ C2β Vps34	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

PI3Kシグナル伝達経路

PI3K阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
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	25766633
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	21882832
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	21882832
human LNCAP cells	Proliferation assay		3 days	Antiproliferative activity against human LNCAP cells after 3 days by MTS assay, IC50=0.21 μM	20227881
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	21882832
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	21882832
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	23795239
human DMS114 cells	Growth inhibition assay			Growth inhibition of human DMS114 cells	22336246
human MKN74 cells	Growth inhibition assay			Growth inhibition of human MKN74 cells	22336246
human SNB78 cells	Growth inhibition assay			Growth inhibition of human SNB78 cells	22336246
human St-4 cells	Growth inhibition assay			Growth inhibition of human St-4 cells	22336246
human DU145 cells	Growth inhibition assay			Growth inhibition of human DU145 cells	22336246
human LOXIMVI cells	Growth inhibition assay			Growth inhibition of human LOXIMVI cells	22336246
human PC3 cells	Growth inhibition assay			Growth inhibition of human PC3 cells	22336246
human LOXIMVI cells	Growth inhibition assay			Growth inhibition of human LOXIMVI cells	22336246
human OVCAR3 cells	Growth inhibition assay			Growth inhibition of human OVCAR3 cells	22336246
human SKOV3 cells	Growth inhibition assay			Growth inhibition of human SKOV3 cells	22336246
human KM12 cells	Growth inhibition assay			Growth inhibition of human KM12 cells	22336246
human HT-29 cells	Growth inhibition assay			Growth inhibition of human HT-29 cells	22336246
human HCT15 cells	Growth inhibition assay			Growth inhibition of human HCT15 cells	22336246
human NCI-H226 cells	Growth inhibition assay			Growth inhibition of human NCI-H226 cells	22336246
human NCI-H522 cells	Growth inhibition assay			Growth inhibition of human NCI-H522 cells	22336246
human A549 cells	Growth inhibition assay			Growth inhibition of human A549 cells	22336246
human HCC2998 cells	Growth inhibition assay			Growth inhibition of human HCC2998 cells	22336246
human SNB75 cells	Growth inhibition assay			Growth inhibition of human SNB75 cells	22336246
human OVCAR4 cells	Growth inhibition assay			Growth inhibition of human OVCAR4 cells	22336246
human OVCAR5 cells	Growth inhibition assay			Growth inhibition of human OVCAR5 cells	22336246
human OVCAR8 cells	Growth inhibition assay			Growth inhibition of human OVCAR8 cells	22336246
human SKOV3 cells	Growth inhibition assay			Growth inhibition of human SKOV3 cells	22336246
human ACHN cells	Growth inhibition assay			Growth inhibition of human ACHN cells	22336246
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生物活性

製品説明

ZSTK474 inhibits class I PI3K isoforms with IC50 of 37 nM in a cell-free assay, mostly PI3Kδ. Phase1/2.

特性

First orally administered PI3K inhibitor used in vivo.

Targets

PI3Kδ ^[2] (Cell-free assay)	PI3Kα ^[2] (Cell-free assay)	PI3K ^[1] (Cell-free assay)	PI3Kβ ^[2] (Cell-free assay)	PI3Kγ ^[2] (Cell-free assay)
4.6 nM	16 nM	37 nM	44 nM	49 nM

In Vitro
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. ZSTK474 inhibits the activities of recombinant p110β, -γ, and -δ with IC50 of 17 nM, 53 nM, and 6 nM, respectively. ZSTK474 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. ZSTK474 treatment at 1 μM blocks membrane ruffling and generation of PIP3 induced by platelet-derived growth factor in murine embryonic fibroblasts (MEFs). ZSTK474 at 10 μM induces apoptosis in OVCAR3 cells, and induces complete G1-phase arrest but not apoptosis in A549 cells. ZSTK474 treatment at 0.5 μM significantly decreases the level of phosphorylated Akt and GSK-3β, as well as the cyclin D1 protein expression. ZSTK474 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. ^[1] ZSTK474 does not inhibit mTOR at 0.1 μM, and even at a concentration of 100 μM, ZSTK474 inhibits mTOR activity less than 40%. ^[2] ZSTK474 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. ^[3] ZSTK474 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). ^[6]
Kinase Assay	Inhibition of PI3K activity
Kinase Assay	A549 cells are lysed in a buffer containing 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 5 mM EDTA, and 1% Igepal CA-630, the lysates are centrifuged at 20,000 g and 4 °C for 10 minutes, and the supernatants are used as cell lysate (protein = 2-4 mg/mL). To immunoprecipitate PI3K, 200 μL of cell lysate are incubated with anti-p85 polyclonal antibody and protein G-agarose (5 μL). PI3Kα, PI3Kβ, and PI3Kδ can be immunoprecipitated by the anti-p85 polyclonal antibody. Agarose beads containing immunoprecipitates are washed twice with buffer A (20 mM Tris-HCl at pH 7.5, 150 mM NaCl, 5 mM EDTA, and 1% Igepal CA-630), once with buffer B (500 mM LiCl and 100 mM Tris-HCl at pH 7.5), once with distilled water, and once with buffer C (100 mM NaCl and 20 mM Tris-HCl at pH 7.5). Immunoprecipitates are suspended in 20 μL of buffer C containing phosphatidylinositol of 200 μg/mL. The mixture is preincubated with increasing concentrations of ZSTK474 at 25 °C for 5 minutes. [γ-³²P]ATP (2 μCi per assay mixture; final concentration, 20 μM) and MgCl₂ (final concentration, 20 mM) are added to start the reaction. The reaction mixture is incubated at 25 °C for 20 minutes. Phosphorylated products of phosphatidylinositol are separated by thin-layer chromatography and visualized by autoradiography. The phosphatidylinositol-3-phosphate region is scraped from the plate, and radioactivity is also measured with liquid scintillation spectroscopy. The level of inhibition for ZSTK474 is determined as the percentage of ³²P counts per minute obtained without ZSTK474.
細胞実験	細胞株	MCF-7, HT-29, HCT-116, OVCAR3, A549, et al.
	濃度	Dissolved in DMSO, final concentrations ~10 μM
	反応時間	48 hours
	実験の流れ	Cells are exposed to increasing concentrations of ZSTK474 for 48 hours. The inhibition of cell proliferation is assessed by measuring changes in total cellular protein by use of a sulforhodamine B assay. Apoptosis is assessed by chromatin condensation or by flow cytometry. For chromatin condensation assay, cells are stained with Hoechst 33342 and examined by fluorescence microscopy. Morphologic changes induced by ZSTK474, such as the condensation of chromatin, are indicative of apoptosis. For flow cytometry analysis, cells are harvested, washed with ice-cold PBS, and fixed in 70% ethanol. Cells are then washed twice with ice-cold PBS again, treated with RNase A (500 μg/mL) at 37 °C for 1 hour, and stained with propidium iodide (25 μg/mL). The DNA content of the cells is analyzed with a flow cytometer.
実験結果図	Methods	Biomarkers	結果図	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

In Vivo
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 irinotecan, cisplatin, doxorubicin, and 5-fluorouracil at their respective maximum tolerable doses with tumor regression of 96%, 35.7%, 24%, or 68.3%, respectively. ZSTK474 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. ^[1] ZSTK474 significantly inhibits tumor growth in the RXF-631L xenograft model, correlated with a significantly reduced number of microvessels in the ZSTK474-treated mice. ^[3] Oral administration of ZSTK474 ameliorates the progression of adjuvant-induced arthritis (AIA) in rats. ^[6]
動物実験	動物モデル	Male BDF1 mice injected subcutaneously with B16F10 cells, and female BALB/c nude mice inoculated subcutaneously with A549, PC-3, or WiDr cells
	投与量	~400 mg/kg/day
	投与経路	Orally

参考
[1] Yaguchi S, et al. J Natl Cancer Inst, 2006, 98(8), 545-556. [2] Kong D, et al. Cancer Sci, 2007, 98(10), 1638-1642. [3] Kong D, et al. Eur J Cancer, 2009, 45(5), 857-865. [4] Marone R, et al. Mol Cancer Res, 2009, 7(4), 601-613. [5] Yang S, et al. PLoS One, 2011, 6(10), e26343. [6] Haruta K, et al. Inflamm Res, 2012, 61(6), 551-562. + 展開

参考

+ 展開

化学情報

分子量	417.41	化学式	C₁₉H₂₁F₂N₇O₂
CAS No.	475110-96-4	SDF	Download ZSTK474 SDFをダウンロードする
Smiles	C1COCCN1C2=NC(=NC(=N2)N3C4=CC=CC=C4N=C3C(F)F)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 : 15 mg/mL ( (35.93 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。)

Water : Insoluble

Ethanol : 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):