Gianni D, Nicolas N, Zhang H, Der Mardirossian C, Kister J, Martinez L, Ferguson J, Roush WR, Brown SJ, Bokoch GM, Hodder P, Rosen H
Department of Immunology and Microbial Science and Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037
Department of Ophthalmology, University of California, San Francisco, San Francisco, CA 94122
The NADPH oxidase (NOX) family catalyzes the regulated formation of reactive oxygen species (ROS). ROS generated via NOX1 have been reported to play a role in a growing number of diseases, including cancer, atherosclerosis, hypertension, neurological disorders and inflammation. Since ROS are also produced by other cellular enzymes, the ability to selectively inhibit NOX1 can be expected to provide reversible, mechanistic insights into these cellular processes with which it is involved. The Scripps Research Institute Molecular Screening Center (SRIMSC), part of the Molecular Libraries Probe Production Centers Network (MLPCN), identified a potent and selective phenothiazine NOX1 inhibitor probe, ML171, by high-throughput screening using a cell-based luminescence assay. ML171 is a potent and selective inhibitor of NOX1, with an IC of 129–156 nM in cell-based assays. ML171 is not cytotoxic, and is selective among NOX family members NOX2, NOX3, and NOX4, as well as against xanthine oxidase, another cellular source of ROS. In addition, ML171 is highly effective in inhibiting the cellular production of invadopodia in a human colon cancer cell line. These results elucidate the relevance of NOX1-dependent ROS generation in mechanisms of cancer invasion, and define compound ML171 as a powerful NOX1 chemical probe and a potential therapeutic agent for treatment of this pathology.
烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶（NOX）家族催化活性氧（ROS）的调控生成。据报道，通过NOX1产生的ROS在越来越多的疾病中发挥作用，包括癌症、动脉粥样硬化、高血压、神经紊乱和炎症。由于ROS也由其他细胞酶产生，因此选择性抑制NOX1的能力有望为其参与的这些细胞过程提供可逆的、基于机制的见解。斯克里普斯研究所分子筛选中心（SRIMSC）是分子文库探针生产中心网络（MLPCN）的一部分，通过基于细胞的发光测定法进行高通量筛选，鉴定出一种强效且选择性的吩噻嗪类NOX1抑制剂探针ML171。ML171是一种强效且选择性的NOX1抑制剂，在基于细胞的测定中IC50为129 - 156 nM。ML171没有细胞毒性，并且在NOX家族成员NOX2、NOX3和NOX4之间具有选择性，同时对ROS的另一个细胞来源黄嘌呤氧化酶也有选择性。此外，ML171在抑制人结肠癌细胞系中侵袭伪足的细胞生成方面非常有效。这些结果阐明了NOX1依赖性ROS生成在癌症侵袭机制中的相关性，并将化合物ML171定义为一种强大的NOX1化学探针和治疗这种病理状况的潜在治疗剂。
