Yuan Ying-Hui, Wang Chen-Chen, Duan Ji-Long, Zhang Peng-Peng, He Meng-Lan, Liu Zhen, Hui Zi, Ye Xiang-Yang
School of Pharmacy, Hangzhou Normal University, Hangzhou, 311121, Zhejiang, China.
Key Laboratory of Elemene Class Anti-Cancer Chinese Medicines; Engineering Laboratory of Development and Application of Traditional Chinese Medicines; Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, 311121, Zhejiang, China.
Mol Divers. 2025 Jul 20. doi: 10.1007/s11030-025-11282-8.
Ataxia telangiectasia mutated and rad3-related kinase (ATR) and histone deacetylases (HDACs) are important therapeutic targets for cancer. In this study, a series of novel ATR/HDACs dual inhibitors containing 2-aminopyrazine motifs were designed and synthesized for the first time. Compounds 18b and 18c not only exhibited good dual inhibition activity against ATR (IC = 6 and 3 nM) and HDAC6 (IC = 6 and 20 nM), but also showed good anti-proliferative activity against four cancer cell lines. Preliminary mechanistic studies showed that compared with 18b, compound 18c exhibited stronger inhibitory effect on tumor cell proliferation and more effective induction of cell death. 18c could induce an increase in the level of acetylated α-tubulin and reduce the phosphorylation levels of ATR (p-ATR) and CHK1 (p-CHK1). Additionally, 18c also demonstrated the ability to upregulate the expression level of γ-H2AX. This research firstly presents the design and synthesis of ATR/HDAC6 dual-target inhibitors. The mechanism of action of these inhibitors may involve the synergistic induction of tumor cell apoptosis by dual inhibition of the DNA damage repair pathway and tubulin deacetylation processes. Overall, the research findings in this paper lay a foundation for further exploiting ATR/HDACs dual inhibitors' therapeutic usefulness while addressing potential drug resistance and adverse effect issues caused by combination therapy in clinical practice.
共济失调毛细血管扩张症突变和rad3相关激酶（ATR）以及组蛋白脱乙酰酶（HDACs）是癌症重要的治疗靶点。在本研究中，首次设计并合成了一系列含有2-氨基吡嗪基序的新型ATR/HDACs双重抑制剂。化合物18b和18c不仅对ATR（IC = 6和3 nM）和HDAC6（IC = 6和20 nM）表现出良好的双重抑制活性，而且对四种癌细胞系也显示出良好的抗增殖活性。初步机制研究表明，与18b相比，化合物18c对肿瘤细胞增殖的抑制作用更强，诱导细胞死亡更有效。18c可诱导乙酰化α-微管蛋白水平升高，并降低ATR（p-ATR）和CHK1（p-CHK1）的磷酸化水平。此外，18c还表现出上调γ-H2AX表达水平的能力。本研究首次报道了ATR/HDAC6双重靶向抑制剂的设计与合成。这些抑制剂的作用机制可能涉及通过双重抑制DNA损伤修复途径和微管蛋白脱乙酰化过程协同诱导肿瘤细胞凋亡。总体而言，本文的研究结果为进一步开发ATR/HDACs双重抑制剂的治疗用途奠定了基础，同时解决了临床实践中联合治疗引起的潜在耐药性和不良反应问题。
