Abstract：
NUAK family kinase 2 (NUAK2) is a promising target for cancer therapeutics due to its reported role in protein phosphorylation, a critical process in cancer cell survival, proliferation, invasion, and senescence. This study aimed to identify novel inhibitors that disrupt NUAK2 activity. We have already identified two KRICT Hippo kinase inhibitor (KHKI) compounds, such as KHKI-01128 and KHKI-01215. Our aim was to evaluate the impact of KHKI-01128 and KHKI-01215 on NUAK2 activity and elucidate its mechanism in colorectal cancer cells.
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.
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.
摘要：
目的：NUAK家族激酶2（NUAK2）由于其在蛋白质磷酸化中的作用而被报道为癌症治疗的有希望的靶标，癌细胞生存的关键过程，扩散,入侵,和衰老。本研究旨在鉴定破坏NUAK2活性的新型抑制剂。我们已经确定了两种KRICT河马激酶抑制剂（KHKI）化合物，如KHKI-01128和KHKI-01215。我们的目的是评估KHKI-01128和KHKI-01215对NUAK2活性的影响，并阐明其在结直肠癌细胞中的作用机制。
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。
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。
