Background
WYE-354 is a powerful and selective ATP-competitive kinase heterocyclic inhibitors with biochemical IC50 of 5 nM against mTOR.[1]
WYE-354 displays very specific selectivity to PI3K families (>100-fold to PI3K? and >500-fold to PI3K?), but it can’t inhibit some protein kinases. May cancer cell lines including MDA-MB-468 and U87MG can be inhibited by WYE-354 in 0.3–1 mM range.[1]
In-vitro study has proved that WYE-354 can inhibit substrate phosphorylation such as p-4E-BP1 T37/46 and p-Akt S473 by mTORC1 and mTORC2. WYE-354 can reduce Akt downstream function and block the propagation of the cancer cell lines with an IC50 value from sub-micromolar to micromolar range [3]. These effects were consistent with the G1 cell cycle arrest in two cell lines, including rapamycin-sensitive and rapamycin-resistant cells. It is believed to induce apoptosis, repress the global protein synthesis, and down-regulate many angiogenic factors.[3]
参考文献:
[1] Qingsong Liu, Carson Thoreen, Jinhua Wang, David Sabatini, Nathanael S. Gray. mTOR mediated anti-cancer drug discovery. Drug Discovery Today. 2009. 6(2): 47-55.
[2] Shi-Yong Sun. mTOR kinase inhibitors as potential cancer therapeutic drugs. Cancer Letters. 28 October 2013. 340(1): 1-8.
Protocol
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Cell experiment [1]:
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Cell lines
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HEK293 cells
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Preparation method
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The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while.Stock solution can be stored below -20°C for several months.
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Reaction Conditions
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1 h; 5 μM
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Applications
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To test directly the inhibition of mTORC2 catalytic activity in vitro, we immunoprecipitated mTORC2 and mTORC1 from HEK293 cells and performed immune-complex kinase assay of the mTORC2-specific substrate His6-AKT or the mTORC1 substrate His6-S6K. AKT (S473) phosphorylation was dose dependently inhibited by WYE-354.
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Animal experiment [1]:
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Animal models
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BALB/c nu/nu female mice
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Dosage form
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50 mg/kg; intraperitoneal injection
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Applications
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Nude mice bearing the PTEN-null PC3MM2 tumors were administered i.p. with vehicle or 50 mg/kg WYE-354.Tumor lysates prepared at 1, 2, 4, and 6 hours after dosing were immunoblotted for levels of P-S6K(T389), P-AKT(S473), and P-AKT(T308). Quantification of the immunoblotting results indicated that WYE-354 completely inhibited P-S6K (T389) for at least 6 hours and substantially inhibited P-AKT(S473) for 6 hour. As expected, P-AKT(T308) in the same tumors was variably but not significantly inhibited. WYE-354 selectively inhibited P-AKT(S473) via targeting mTORC2.
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Other notes
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Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.
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参考文献:
[1] Yu K, Toral-Barza L, Shi C, et al. Biochemical, cellular, and in vivo activity of novel ATP-competitive and selective inhibitors of the mammalian target of rapamycin[J]. Cancer research, 2009, 69(15): 6232-6240.
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