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    InvivoChem Cat #: V0193
    CAS #: 1062169-56-5Purity ≥98%

    Description: WYE-354 (WYE 354) is a novel, potent, specific, cell-permeable and ATP-competitive mTOR (mammalian target of rapamycin) inhibitor with potential anticancer/antitumor activity. It inhibits mTOR with IC50 of 5 nM.

    References:  Cancer Res. 2009;69(15):6232-40; Biochem Biophys Res Commun. 2011 Apr 22;407(4):714-9.  

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    Molecular Weight (MW)




    CAS No.



    -20℃ for 3 years in powder form

    -80℃ for 2 years in solvent

    Solubility (In vitro)

    DMSO: 99 mg/mL (199.8 mM)

    Water: <1 mg/mL

    Ethanol: <1 mg/mL

    Solubility (In vivo)

    4% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/mL 

    Synonyms & Other info

    WYE354; WYE 354; WYE-354;

    Chemical Name: methyl 4-(6-(4-((methoxycarbonyl)amino)phenyl)-4-morpholino-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate


    InChi Code: InChI=1S/C24H29N7O5/c1-34-23(32)26-17-5-3-16(4-6-17)20-27-21(29-11-13-36-14-12-29)19-15-25-31(22(19)28-20)18-7-9-30(10-8-18)24(33)35-2/h3-6,15,18H,7-14H2,1-2H3,(H,26,32)


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    In Vitro

    Kinase Assay: The assays are performed in 96-well plates for 2 hours at room temperature in 25 μL containing 6 nM Flag-TOR(3.5), 1 μM His6-S6K, and 100 μM ATP. The assays are performed and detected by DELFIA employing the Eu-phospho-p70S6K T389 antibody. For inhibitor versus ATP matrix competition, mTOR kinase reactions are carried out with varying concentrations of ATP (0, 25, 50 100, 200, 400, and 800 μM) in combination with varying concentrations of WYE-354. The assays contained 12 nM Flag-TOR(3.5), 1 μM His-S6K, and are incubated for 30 min. The assay results are similarly detected by DELFIA and processed for generation of double-reciprocal plots.


    Cell Assay: Cells (Tumor cell lines including MDA-MB-361, MDA-MB-231, MDA-MB-468, LNCap, DU145, A498, and HCT116) are plated in 96-well plates at 1000 to 3000 cells per well for 24 hours, treated with DMSO or varying concentrations of WYE-354. Viable cell densities are determined 72 hours later by MTS assay employing a CellTiter 96 kit. The effect of each treatment is calculated as percent of control growth relative to the DMSO-treated cells grown in the same culture plate. Inhibitor dose response curves are plotted for determination of IC50 values.

    WYE-354 also inhibits several PI3Ks at micromolar levels. In HEK293 cells, WYE-354 (0.2 μM–5 μM) effectively inhibits both mTORC1 and mTORC2. WYE-354 (0.3 μM–10 μM) significantly blocks mTOR signaling and Akt activation in U87MG and MDA361 cells. Furthermore, WYE-354 potently inhibits proliferation in tumor cell lines including MDA-MB-361, MDA-MB-231, MDA-MB-468, LNCap, A498, and HCT116, with IC50 values ranging from 0.28 μM to 2.3 μM. The apoptosis induced by WYE-354 is accompanied by G1 cell cycle arrest and caspases activation. In endothelial HUVEC cells, WYE-354 (10 nM–1 μM) also inhibits both mTORC1 and mTORC2 signaling, as revealed by dephosphorylation of S6 ribosomal protein and Akt, respectively. Furthermore, WYE-354 (10 nM–1 μM) activates mitogen-activated protein kinase (MAPK) signaling, which may be due to its inhibition of mTORC1.

    In Vivo

    In a mice xenograft model of PTEN-null PC3MM2 tumor, WYE-354 (50 mg/kg) effectively inhibits mTOR signaling and tumor growth.

    Animal model

    Nude mice (BALB/c, nu/nu, female) bearing PC3MM2 xenograft

    Formulation & Dosage

    Formulated in 5% ethanol, 5% polysorbate 80, 5% PEG-400; 50 mg/kg; i.p. injection.


    Cancer Res. 2009;69(15):6232-40; Biochem Biophys Res Commun. 2011 Apr 22;407(4):714-9.  

    These protocols are for reference only. InvivoChem does not independently validate these methods.


    In vivo efficacy of WYE-354 on human gallbladder cancer xenografts.  2015 Oct 13;6(31):31877-88.


    In vivo efficacy of rapamycin on human gallbladder cancer xenografts.  2015 Oct 13;6(31):31877-88.


    In vitro effects of WYE-354 on cell growth and mTOR signaling pathway in two gallbladder cancer cell lines.  2015 Oct 13;6(31):31877-88.


    Novel pyrazolopyrimidines are ATP-competitive inhibitors of mTOR. Cancer Res. 2009 Aug 1;69(15):6232-40


    mTOR inhibitors down-regulate angiogenic factors.


    WYE-354 inhibits mTOR signaling and tumor growth in vivo.


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