Small-molecule inhibitor of USP7/HAUSP ubiquitin protease stabilizes and activates p53 in cells
Small-molecule inhibitor of USP7/HAUSP ubiquitin protease stabilizes and activates p53 in cells
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DOI：
10.1158/1535-7163.mct-09-0097
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发表时间：
2009-08-01
影响因子：
5.7
通讯作者：
Daviet, Laurent
中科院分区：
文献类型：
--
作者：
Colland, Frederic;Formstecher, Etienne;Daviet, Laurent
泛素/蛋白酶体系统的失调与包括癌症在内的许多人类疾病的发病机制有关。泛素特异性蛋白酶（USP）是参与蛋白质底物的去泛素化的半胱氨酸蛋白酶。USP 7与必需病毒蛋白和致癌途径（如p53/Mdm 2和磷脂酰肌醇3-激酶/蛋白激酶B网络）之间的功能联系强烈表明，用小分子抑制剂靶向USP 7可能有助于治疗癌症和病毒性疾病。使用高通量筛选，我们发现了HBX 41，108，一种抑制USP 7去泛素化活性的小分子化合物，IC 50在亚微摩尔范围内。动力学数据表明非竞争性可逆抑制机制。HBX 41，108在体外和细胞中显示影响USP 7介导的p53去泛素化。作为癌细胞中RNA干扰介导的USP 7沉默，HBX 41，108处理稳定了p53，激活了p53靶基因的转录而不诱导遗传毒性应激，并抑制了癌细胞生长。最后，HBX 41，108诱导p53依赖性细胞凋亡，如p53野生型和无效等基因癌细胞系所示。因此，我们报告了第一个针对USP 7的铅样抑制剂的鉴定，为开发新的抗癌药物提供了结构基础。[Mol Cancer Ther 2009;8（8）：2286-95]
Deregulation of the ubiquitin/proteasome system has been implicated in the pathogenesis of many human diseases, including cancer. Ubiquitin-specific proteases (USP) are cysteine proteases involved in the deubiquitination of protein substrates. Functional connections between USP7 and essential viral proteins and oncogenic pathways, such as the p53/Mdm2 and phosphatidylinositol 3-kinase/protein kinase B networks, strongly suggest that the targeting of USP7 with small-molecule inhibitors may be useful for the treatment of cancers and viral diseases. Using high-throughput screening, we have discovered HBX 41,108, a small-molecule compound that inhibits USP7 deubiquitinating activity with an IC50 in the submicromolar range. Kinetics data indicate an uncompetitive reversible inhibition mechanism. HBX 41,108 was shown to affect USP7-mediated p53 deubiquitination in vitro and in cells. As RNA interference-mediated USP7 silencing in cancer cells, HBX 41,108 treatment stabilized p 53, activated the transcription of a p53 target gene without inducing genotoxic stress, and inhibited cancer cell growth. Finally, HBX 41,108 induced p53-dependent apoptosis as shown in p53 wild-type and null isogenic cancer cell lines. We thus report the identification of the first lead-like inhibitor against USP7, providing a structural basis for the development of new anticancer drugs. [Mol Cancer Ther 2009;8(8):2286-95]