To evaluate anticancer properties of these inhibitors, four in vitro assays in the SW480 cell line (time-resolved fluorescence resonance energy transfer assay, KINOMEscan kinase profiling, viability, and apoptosis assays) and two pharmacological mechanism analyses (Gene Set Enrichment Analysis and western blotting) were performed.
KHKI-01128 and KHKI-01215 exhibited potent inhibitory activity against NUAK2 (half-maximal inhibitory concentration=0.024±0.015 μM and 0.052±0.011 μM, respectively). These inhibitors suppressed cell proliferation, with half-maximal inhibitory concentrations of 1.26±0.17 μM and 3.16±0.30 μM, respectively, and induced apoptosis of SW480 cells. Gene Set Enrichment Analysis revealed negative enrichment scores of -0.84 for KHKI-01128 (false-discovery rate=0.70) and 1.37 for KHKI-01215 (false-discovery rate=0.18), indicating that both effectively suppressed the expression of YES1-associated transcriptional regulator (YAP) target genes.
These results suggest that KHKI-01128 and KHKI-01215 are potent NUAK2 inhibitors with promising potential for pharmaceutical applications.
方法:为了评估这些抑制剂的抗癌特性,SW480细胞系中的四种体外测定(时间分辨荧光共振能量转移测定,KINOMEscan激酶谱,生存能力,和凋亡测定)和两个药理学机制分析(基因集富集分析和蛋白质印迹)。
结果:KHKI-01128和KHKI-01215对NUAK2表现出有效的抑制活性(半最大抑制浓度=0.024±0.015μM和0.052±0.011μM,分别)。这些抑制剂抑制细胞增殖,半最大抑制浓度为1.26±0.17μM和3.16±0.30μM,分别,诱导SW480细胞凋亡。基因集富集分析显示,KHKI-01128的负富集评分为-0.84(错误发现率=0.70),KHKI-01215的负富集评分为1.37(错误发现率=0.18),这表明两者都有效地抑制了YES1相关转录调节因子(YAP)靶基因的表达。
结论:这些结果表明,KHKI-01128和KHKI-01215是有效的NUAK2抑制剂,具有潜在的药物应用潜力。