Université de Lyon, INSERM U1052, Centre de Recherche en Cancérologie de Lyon, Lyon, France.
Mol Pharmacol. 2013 Jul;84(1):12-24. doi: 10.1124/mol.112.082347. Epub 2013 Apr 11.
The benefit of cancer chemotherapy based on alkylating agents is limited because of the action of DNA repair enzymes, which mitigate the damage induced by these agents. The interaction between the proteins ERCC1 and XPF involves two major components of the nucleotide excision repair pathway. Here, novel inhibitors of this interaction were identified by virtual screening based on available structures with use of the National Cancer Institute diversity set and a panel of DrugBank small molecules. Subsequently, experimental validation of the in silico screening was undertaken. Top hits were evaluated on A549 and HCT116 cancer cells. In particular, the compound labeled NSC 130813 [4-[(6-chloro-2-methoxy-9-acridinyl)amino]-2-[(4-methyl-1-piperazinyl)methyl]] was shown to act synergistically with cisplatin and mitomycin C; to increase UVC-mediated cytotoxicity; to modify DNA repair as indicated by the staining of phosphorylated H2AX; and to disrupt interaction between ERCC1 and XPF in cells. In addition, using the Biacore technique, we showed that this compound interacts with the domain of XPF responsible for interaction with ERCC1. This study shows that small molecules targeting the protein-protein interaction of ERCC1 and XPF can be developed to enhance the effects of alkylating agents on cancer cells.
基于烷化剂的癌症化疗的益处是有限的,因为 DNA 修复酶的作用会减轻这些药物引起的损伤。ERCC1 和 XPF 蛋白之间的相互作用涉及核苷酸切除修复途径的两个主要组成部分。在这里,我们使用国家癌症研究所多样性集和药物银行小分子面板,通过基于现有结构的虚拟筛选,鉴定了这种相互作用的新型抑制剂。随后,对计算机筛选进行了实验验证。在 A549 和 HCT116 癌细胞上对顶级命中物进行了评估。特别是,被标记为 NSC 130813 [4-[(6-氯-2-甲氧基-9-吖啶基)氨基]-2-[(4-甲基-1-哌嗪基)甲基]]的化合物被证明与顺铂和丝裂霉素 C 具有协同作用;增加 UVC 介导的细胞毒性;如磷酸化 H2AX 的染色所示,改变 DNA 修复;并破坏细胞中 ERCC1 和 XPF 之间的相互作用。此外,使用 Biacore 技术,我们表明该化合物与 XPF 负责与 ERCC1 相互作用的结构域相互作用。这项研究表明,针对 ERCC1 和 XPF 蛋白-蛋白相互作用的小分子可以被开发出来,以增强烷化剂对癌细胞的作用。