WZ4002 is a mutant-selective EGFR inhibitor for EGFR(L858R) and EGFR(T790M) with IC50 of 2 nM/8 nM.

EGFR (L858R/T790M):8 nM, EGFR (L858R):2 nM

WZ4002对其他EGFR基因型E746_A750和E746_A750/T790M的抑制作用IC50分别为2 nM和6 nM。此外，WZ4002能以IC50为32 nM抑制野生型ERBB2。WZ4002在NSCLC细胞系中抑制EGFR、AKT和ERK1/2的磷酸化，并阻止表达不同EGFR T790M突变等位基因的NIH-3T3细胞中EGFR的磷酸化。WZ4002 在10 μM时相对于DMSO对照组抑制效果大于95%的激酶来测量它们的解离常数。WZ4002在其C2-苯胺取代基上具有一个邻甲氧基。与WZ3146相比，WZ4002 对EGFR的选择性更强。与喹唑啉抑制剂相比，WZ4002抑制WT EGFR磷酸化的效力降低了100倍。同样，WZ4002对重组L858R/T790M蛋白的EGFR激酶活性的抑制作用比对WT EGFR更为强烈，而HKI-272和吉非替尼则观察到相反的情况。[1] 另外，第三代EGFR TKI WZ4002能完全抑制Src TKI耐药的H1975细胞和HCC827细胞中磷酸化的EGFR。[2]

在为期两周的有效性研究中，与单独使用载体相比，WZ4002治疗在含有T790M的小鼠模型中显著促进了肿瘤回缩。[1] 低剂量WZ4002和高剂量WZ4002治疗分别使示踪剂吸收平均降低了26%和36%。[3]

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.

The NSCLC, Ba/F3 cells, NIH-3T3 cells, PC9 gR4 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.(Only for Reference)