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Research Area
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Description
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Cancer |
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Biological Activity
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Description
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WZ4002 is a novel, mutant-selective EGFR inhibitor for EGFRL858R and EGFRL858R/T790M with IC50 of 2 nM and 8 nM, respectively. |
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Targets
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EGFRL858R |
EGFRL858R/T790M |
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IC50 |
2 nM |
8 nM [1] |
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In Vitro
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WZ4002 inhibits other EGFR genotypes E746_A750 and E746_A750/T790M with IC50 of 2 and 6 nM. Besides, WZ4002 suppresses widetype ERBB2 with an IC50 of 32 nM. WZ4002 inhibits EGFR, AKT and ERK1/2 phosphorylation in NSCLC cell lines and WZ4002 prevents of EGFR phosphorylation in NIH-3T3 cells expressing different EGFR T790M mutant alleles. For WZ4002, kinases that exhibited greater than 95% inhibition relative to the DMSO control at 10 μM are selected for measurement of their dissociation constants. WZ4002, which possesses an ortho-methoxy group at the C2-aniline substituent, is more selective for EGFR compared to WZ3146. WZ4002 is 100-fold less effective at inhibiting phosphorylation of WT EGFR compared to the quinazoline inhibitors. Similarly, WZ4002 prevents EGFR kinase activity of recombinant L858R/T790M protein more potently than of WT EGFR, while the opposite is observed with HKI-272 and gefitinib. [1] In addition, the phosphorylated EGFR of Src TKI-resistant H1975 cells, as well as HCC827 cells, is completely suppressed by the third generation EGFR TKI, WZ4002. [2] |
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In Vivo
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In a 2-week efficacy study, WZ4002 treatment results in significant tumor regressions compared to vehicle alone in both T790M containing murine models. [1] Treatment with low-dose WZ4002, and high-dose WZ4002 leads to mean decreases in tracer uptake of 26%, and 36%, respectively. [3] |
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Clinical Trials
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Features
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Protocol
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Kinase Assay
[1]
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| EGFR kinase assays |
In vitro inhibitory enzyme kinetic assays using recombinant EGFR L858R/T790M and WT protein and are performed using the ATP/NADH coupled assay system in a 96-well format. WZ4002 is added to determine its inhibitory effects. |
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Cell Assay
[1]
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| Cell Lines |
NSCLC, Ba/F3 cells, NIH-3T3 cells, PC9GR4 cells |
| Concentrations |
0-1 μM |
| Incubation Time |
72 hours |
| Methods |
The NSCLC, Ba/F3 cells, NIH-3T3 cells, PC9GR4 cells are used and verified to contain EGFR delE746_A750/T790M by direct sequencing. Cell proliferation and growth assays are performed using the MTS assay. Site directed mutagenesis is performed using the Quick Change Site-Directed Mutagenesis kit. |
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Animal Study
[1]
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| Animal Models |
EGFR-TL (T790M/L858R) mice |
| Formulation |
NMP (10% 1-methyl-2-pyrrolidinone: 90% PEG-300 |
| Doses |
25mg/kg |
| Administration |
Gavage |
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