WYE-354

 

Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

Biological Activity

Description	WYE-354 is a potent, specific and ATP-competitive inhibitor of mTOR with IC50 of 5 nM, blocks mTORC1/P-S6K(T389) and mTORC2/P-AKT(S473) not P-AKT(T308), selective for mTOR than PI3Kα (>100-fold) and PI3Kγ (>500-fold).
Targets	mTOR	PI3Kα	PI3Kγ
IC50	5 nM	1.89 μM	7.37 μM [1]
In vitro	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. [1] 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. [2]
In vivo	In a mice xenograft model of PTEN-null PC3MM2 tumor, WYE-354 (50 mg/kg) effectively inhibits mTOR signaling and tumor growth. [1]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

mTOR inhibitor assays

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: ^[1]

Cell lines	Tumor cell lines including MDA-MB-361, MDA-MB-231, MDA-MB-468, LNCap, DU145, A498, and HCT116
Concentrations	0–50 μM, dissolved in DMSO
Incubation Time	72 hours
Method	Cells 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.

Animal Study: ^[1]

Animal Models	Nude mice (BALB/c, nu/nu, female) bearing PC3MM2 xenograft
Dosages	50 mg/kg
Administration	Administered via intraperitoneal injection
Solubility	30% PEG400/0.5% Tween80/5% propylene glycol, 30 mg/mL

1

References

Chemical Information

Download WYE-354 SDF

Chemical Name

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.49553	4.9553	9.9106	14.8659

Customer Reviews (3)

Click to enlarge	Rating
	Source	Dr. Yong-Weon Yi from Georgetown University Medical Center.WYE-354 purchased from Selleck
	Method	MTT assays
	Cell Lines	HCC1937 cells, SUM149PT cells, MDA-MB-231 cells, MDA-MB-463 cells
	Concentrations	0-10 μM
	Incubation Time	72 h
	Results	WYE-354 decreased the viability of HCC1937 cells, SUM149PT cells, MDA-MB-231 cells, and MDA-MB-463 cells in a dose-dependent manner.

Product Citations (2)

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  mTOR regulates phagosome and entotic vacuole fission. [Krajcovic M, et al. Mol Biol Cell 2013;24(23):3736-45]

  
  PubMed: 24088573


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  Inhibition of mTORC1 kinase activates Smads 1 and 5 but not Smad8 in human prostate cancer cells, mediating cytostatic response to rapamycin. [Wahdan-Alaswad RS, et al. Mol Cancer Res 2012;10(6):821-33]

  
  PubMed: 22452883