Department of Urology, Reproductive Medicine and Oncology, The Affiliated Taizhou People's Hospital of Nanjing Medical University, Taizhou, 225300, People's Republic of China.
Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing, 211198, People's Republic of China.
Drug Des Devel Ther. 2024 Nov 20;18:5283-5297. doi: 10.2147/DDDT.S479113. eCollection 2024.
Dual inhibition of ataxia telangiectasia mutated (ATM) and histone deacetylase 2 (HDAC2) may be a potential strategy to improve antitumor efficacy in testicular cancer.
A combined virtual screening protocol including pharmacophore modeling and molecular docking was used for screening potent dual-target ATM/HDAC2 inhibitors. In order to obtain the optimal lead compound, the dual ATM/HDAC2 inhibitory activity of the screened compounds was further evaluated using enzyme inhibition methods. The binding stability of the optimal compound to the dual targets was verified by molecular dynamics (MD) simulation. MTT assay and in vivo antitumor experiment were performed to validate antitumor efficacy of the optimal compound in testicular cancer.
Here, we successfully discovered six potent dual-target ATM/HDAC2 inhibitors (AMHs 1-6), which exhibited good inhibitory activity against both ATM and HDAC2. Among them, AMH-4 showed strong inhibitory activity against both ATM (IC = 1.12 ± 0.03 nM) and HDAC2 (IC = 3.04 ± 0.08 nM). MD simulation indicated that AMH-4 binds to ATM and HDAC2 with satisfactory stability. Importantly, AMH-4 had significant antiproliferative activity on human testicular tumor cells, especially NTERA-2 cL.D1 cells, and no inhibitory effect on normal human testicular cells. In vivo experiments exhibited that AMH-4 was more effective than lartesertib and vorinostat in inhibiting the growth of NTERA-2 cL.D1 xenograft tumors with low toxicity.
Overall, these results suggest that AMH-4 is an effective and low toxicity candidate for the treatment of testicular germ cell tumors.
双重抑制共济失调毛细血管扩张突变（ATM）和组蛋白去乙酰化酶 2（HDAC2）可能是提高睾丸癌抗肿瘤疗效的一种潜在策略。
采用包括药效团模型和分子对接的组合虚拟筛选方案筛选有效的双靶 ATM/HDAC2 抑制剂。为了获得最佳的先导化合物，进一步采用酶抑制法评价筛选化合物对双靶的双重抑制活性。通过分子动力学（MD）模拟验证最佳化合物与双靶的结合稳定性。MTT 检测和体内抗肿瘤实验验证了最佳化合物在睾丸癌中的抗肿瘤功效。
在此，我们成功发现了六种有效的双靶 ATM/HDAC2 抑制剂（AMHs 1-6），它们对 ATM 和 HDAC2 均表现出良好的抑制活性。其中，AMH-4 对 ATM（IC = 1.12 ± 0.03 nM）和 HDAC2（IC = 3.04 ± 0.08 nM）均具有较强的抑制活性。MD 模拟表明 AMH-4 与 ATM 和 HDAC2 结合具有良好的稳定性。重要的是，AMH-4 对人睾丸肿瘤细胞，特别是 NTERA-2 cL.D1 细胞具有显著的增殖抑制活性，而对正常人类睾丸细胞没有抑制作用。体内实验表明，与 lartesertib 和 vorinostat 相比，AMH-4 能更有效地抑制 NTERA-2 cL.D1 异种移植瘤的生长，且毒性较低。
综上所述，这些结果表明 AMH-4 是治疗睾丸生殖细胞肿瘤的有效且低毒候选药物。
