Home PI3K/Akt/mTOR mTOR inhibitor WYE-125132 (WYE-132)

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WYE-125132 (WYE-132) mTOR inhibitor

Cat.No.S2661

WYE-125132 (WYE-132) is a highly potent, ATP-competitive mTOR inhibitor with IC50 of 0.19 nM; it is highly selective for mTOR versus PI3Ks or PI3K-related kinases hSMG1 and ATR.
WYE-125132 (WYE-132) mTOR inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 519.6

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Quality Control

Batch: S266101 DMSO]104 mg/mL]false]Water]Insoluble]false]Ethanol]Insoluble]false Purity: 100%
100

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
LNCAP cells Cytotoxicity assay Cytotoxicity against human LNCAP cells, IC50=0.002 μM
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight 519.6 Formula

C27H33N7O4

 Storage (From the date of receipt)
CAS No. 1144068-46-1 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles CNC(=O)NC1=CC=C(C=C1)C2=NC3=C(C=NN3C4CCC5(CC4)OCCO5)C(=N2)N6CC7CCC(C6)O7

Solubility

In vitro
Batch:

DMSO : 104 mg/mL (200.15 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo
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In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Mechanism of Action

Features
A highly potent, ATP-competitive, and specific mTOR kinase inhibitor.
Targets/IC50/Ki
mTOR [1]
(Cell-free assay)
0.19 nM
In vitro
WYE-125132 (WYE-132) potently and ATP-competitively inhibits recombinant mTOR kinase with IC50 of 0.19 nM and also shows high selectivity over various PI3Ks and a panel of 230 protein kinases. [1] In vitro, it exhibits significant anti-proliferative activity against a panel of tumor cell lines with IC50 ranging from 2 nM (LNCap) to 380 nM (HTC116). Besides, this compound also causes cell cycle progression, induction of apoptosis, and inhibition of protein synthesis and cell size. [1] It results in a significant reduction in the synthesis of pre-tRNALeu by 72%, 80%, and 53% in actively proliferating cells of MG63, MDA361, and HEK293, respectively by inhibiting mTORC1. Moreover, WYE-125132 is also found to induce the dephosphorylation of Maf1 (negative regulator of Pol III transcription) and its accumulation in the nucleus. [2]
Kinase Assay
Kinase assays
mTOR enzyme assays via dissociation-enhanced lanthanide fluorescent immunoassay (DELFIA), ATP matrix assays, and mTOR immune-complex kinase assays are performed as follows for WYE-125132 (WYE-132). The endogenous TOR of LNCap cell lysate is immunoprecipitated by anti-FRAP/TOR (N-19). Cell lysate (1.0 mg) is mixed with 4 μg antibody coupled to protein-G/A agarose in 1 mL lysis buffer. The immune-complexes are washed sequentially with lysis buffer, lysis buffer plus 500 mM KCl, and kinase buffer wash. The immune-complexes are subjected to kinase reaction for 30 minutes at 30 °C in a final volume of 50 μL containing 10 mM Hepes (pH 7.4), 50 mM NaCl, 50 mM β-glycerophosphate, and 0.5 μM microcystin LR, 1 mM DTT, 10 mM MnCl2, 100 μM ATP, 1 μg His6-S6K or 1 μg His6-4EBP1. Kinase reactions (immune-complex and purified enzymes) are terminated by NuPAGE LDS sample buffer and resolved in a 4-12% NuPAGE Bis-Tris gel for Western blotting with anti-P(T389)-p70S6K and anti-P(T46)-4EBP1, anti-FRAP/TOR (N-19), anti-FLAG M2, and anti-His6 (Clone His-1). In the radioactive assay, 10 μCi [γ-32P]ATP (3000 Ci/mmol) and 100 μM cold ATP are used. 32P-labeled products are resolved by SDS-PAGE and subjected to autoradiogram to Kodak X-ray films.
In vivo
WYE-125132 (WYE-132) (5 mg/kg p.o.) produces significant antitumor activity and causes dose-dependent tumor growth delay in the PI3K/mTOR- and HER2-hyperactive MDA361 tumor model. In addition, it also shows potent antitumor efficacy in the PTEN-null glioma U87MG, non-small cell lung cancer H1975 and A549 models. [1]
References

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Frequently Asked Questions

Question 1:
Whether the vehicle “30% PEG400+0.5% Tween80+5% Propylene glycol” you recommended for it is a clear solution or not? What’s the suggested formula for i.p. injection?

Answer:
The formula on our website “30% PEG400+0.5% Tween80+5% Propylene glycol” is not a clear solution and can be used for p.o. administration. For i.p. solution, there are two methods to prepare it: 1）2% DMSO+30% PEG 300+ddH2O, 2.5mg/ml, stable for 1h at RT; 2）2% DMSO+30% PEG 300+ 2% Tween80 +ddH2O, 2.5mg/ml, stable for 2h at RT.

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