Home PI3K/Akt/mTOR AMPK inhibitor WZ4003

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WZ4003 AMPK inhibitor

Cat.No.S7317

WZ4003 is a highly specific NUAK kinase inhibitor with IC50 of 20 nM and 100 nM for NUAK1 and NUAK2 in cell-base assays, respectively, without significant inhibition on 139 other kinases.
WZ4003 AMPK inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 496.99

Jump to Mechanism of Action Solubility Chemical Information, Storage & Stability Specificity

Quality Control

Batch: S731701 DMSO]10 mg/mL]false]Water]Insoluble]false]Ethanol]Insoluble]false Purity: 99.72%
99.72

Chemical Information, Storage & Stability

Molecular Weight 496.99 Formula

C25H29ClN6O3

 Storage (From the date of receipt)
CAS No. 1214265-58-3 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles CCC(=O)NC1=CC(=CC=C1)OC2=NC(=NC=C2Cl)NC3=C(C=C(C=C3)N4CCN(CC4)C)OC

Solubility

In vitro
Batch:

DMSO : 10 mg/mL (20.12 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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Mechanism of Action

Targets/IC50/Ki
NUAK1 [1]
(Cell-free assay)
20 nM
NUAK2 [1]
(Cell-free assay)
100 nM
In vitro
In HEK-293 cells expressing wild-type NUAK1, WZ4003 (3–10 μM) markedly suppresses NUAK1-mediated MYPT1 phosphorylation. Moreover, this compound (10 μM) inhibits MYPT1 Ser445 phosphorylation as well as cell migration, invasion and proliferation to a similar extent as knock out in MEFs or knock down in U2OS cells of NUAK1. [1] It also exhibits a high, specific affinity to the L858R/T790M mutant EGFR, while a significantly reduced cellular IC50 against T790M containing Ba/F3 cells. [2]
Kinase Assay
IC50 determination
Active GST–NUAK1, GST–NUAK1[A195T] and GST–NUAK2 enzymes are purified using glutathione–Sepharose from HEK-293 cell lysates 36–48 h following the transient transfection of pEBG2T mammalian constructs expressing N-terminal GSTtagged NUAK1, NUAK1[A195T] or NUAK2. For peptide kinase assays, 96-well plates are used, and each reaction is performed in triplicate. Each reaction is set up in a total volume of 50 μL containing 100 ng of NUAK1 (wild-type or A195T mutant) or NUAK2 in 50 mM Tris/HCl (pH 7.5), 0.1 mM EGTA, 10 mM magnesium acetate, 200 μM Sakamototide, 0.1 mM [γ -32P]ATP (450–500 c.p.m./pmol) and the indicated concentrations of inhibitors dissolved in DMSO. After incubation for 30 min at 30°C, reactions are terminated by adding 25 mM (final) EDTA to chelate the magnesium. Then, 40 μL of the reaction mix is spotted on to P81 paper and immersed in 50 mM orthophosphoric acid. Samples are washed three times in 50 mM orthophosphoric acid followed by a single acetone rinse and air drying. The incorporation of [γ -32P]ATP into Sakamototide is quantified by Cerenkov counting. The values are expressed as a percentage of the DMSO control. IC50 curves are developed and IC50 values are calculated using GraphPad Prism software.
References

Applications

Methods Biomarkers Images PMID
Western blot Tau-5 / p-tau (Ser356) S7317-WB1 27720485
Immunofluorescence YAP/TAZ S7317-IF1 30158528

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