You Chang, Wei Shijiao, Yu Jia, Xu Guangcan, Li Huimin, Liu Xinyu, Wang Menghan, Meng Xueling, Xu Youyin, Yu Gang, Luo Heng, Xu Bixue
College of Pharmacy, Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China.
State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, Guizhou Medical University, Guiyang, 550014, China.
Mol Divers. 2025 Jun 15. doi: 10.1007/s11030-025-11225-3.
To uncover novel inhibitors of Werner (WRN) helicase, this study adopted the scaffold-hopping strategy to design and synthesize 24 novel 2-amino-4-(trifluoromethyl)pyrimidine derivatives. The MTT assay was employed to evaluate the anticancer activity of target compounds against microsatellite instability-high (MSI-H) cell lines (HCT116 and LNCaP) and microsatellite stability (MSS) cell lines (SW620 and PC3). Some compounds demonstrated significant inhibitory activity against all four cancer cell lines. Specifically, compounds 11c, 11f, 11 g, 11 h, and 11 l exhibited greater inhibitory effect toward MSI-H cells (HCT116 and LNCaP) compared to MSS cells (SW620 and PC3). The most active compound 11 g exhibited excellent cellular selectivity, with IC values of 1.52 and 1.72 μM against MSI-H cell lines (HCT116 and LNCaP), respectively, while the IC values of 11 g against MSS cell lines (SW620 and PC3) were 4.24 and 2.78 μM, respectively, followed by the compound 11 h, whose IC values against HCT116, LNCaP, SW620, and PC3 cell lines were 2.22, 1.6, 2.37, and 3.21 µM, respectively. The results of apoptosis induction and cell cycle arrest experiments indicate that compounds 11 g and 11 h induced early apoptosis in HCT116 cells, and G2/M phase cell cycle arrest. This finding was further validated through molecular docking analysis and cellular thermal migration and enzyme activity experiments. The results of WRN helicase inhibition assays showed that the IC value of compound 11 g was 6.61 µM. In summary, our study identifies 2-amino-4-(trifluoromethyl)pyrimidine derivative 11 g as potential WRN-dependent anticancer agents.
为了发现Werner（WRN）解旋酶的新型抑制剂，本研究采用骨架跃迁策略设计并合成了24种新型2-氨基-4-（三氟甲基）嘧啶衍生物。采用MTT法评估目标化合物对微卫星高度不稳定（MSI-H）细胞系（HCT116和LNCaP）和微卫星稳定（MSS）细胞系（SW620和PC3）的抗癌活性。一些化合物对所有四种癌细胞系均表现出显著的抑制活性。具体而言，与MSS细胞（SW620和PC3）相比，化合物11c、11f、11g、11h和11l对MSI-H细胞（HCT116和LNCaP）表现出更强的抑制作用。活性最高的化合物11g表现出优异的细胞选择性，对MSI-H细胞系（HCT116和LNCaP）的IC值分别为1.52和1.72μM，而11g对MSS细胞系（SW620和PC3）的IC值分别为4.24和2.78μM，其次是化合物11h，其对HCT116、LNCaP、SW620和PC3细胞系的IC值分别为2.22、1.6、2.37和3.21μM。细胞凋亡诱导和细胞周期阻滞实验结果表明，化合物11g和11h诱导HCT116细胞早期凋亡，并导致G2/M期细胞周期阻滞。通过分子对接分析、细胞热迁移和酶活性实验进一步验证了这一发现。WRN解旋酶抑制试验结果表明，化合物11g的IC值为6.61μM。总之，我们的研究确定2-氨基-4-（三氟甲基）嘧啶衍生物11g为潜在的依赖WRN的抗癌药物。
