Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.
Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):4834-9. doi: 10.1073/pnas.1015312108. Epub 2011 Mar 7.
A specific small-molecule inhibitor of p97 would provide an important tool to investigate diverse functions of this essential ATPase associated with diverse cellular activities (AAA) ATPase and to evaluate its potential to be a therapeutic target in human disease. We carried out a high-throughput screen to identify inhibitors of p97 ATPase activity. Dual-reporter cell lines that simultaneously express p97-dependent and p97-independent proteasome substrates were used to stratify inhibitors that emerged from the screen. N2,N4-dibenzylquinazoline-2,4-diamine (DBeQ) was identified as a selective, potent, reversible, and ATP-competitive p97 inhibitor. DBeQ blocks multiple processes that have been shown by RNAi to depend on p97, including degradation of ubiquitin fusion degradation and endoplasmic reticulum-associated degradation pathway reporters, as well as autophagosome maturation. DBeQ also potently inhibits cancer cell growth and is more rapid than a proteasome inhibitor at mobilizing the executioner caspases-3 and -7. Our results provide a rationale for targeting p97 in cancer therapy.
一种特定的 p97 小分子抑制剂将为研究这种与多种细胞活动相关的必需 ATP 酶（AAA ATP 酶）的多种功能提供重要工具，并评估其在人类疾病中作为治疗靶点的潜力。我们进行了高通量筛选，以鉴定 p97 ATP 酶活性的抑制剂。同时表达 p97 依赖性和 p97 非依赖性蛋白酶体底物的双报告细胞系用于分层筛选出的抑制剂。N2，N4-二苄基喹唑啉-2,4-二胺（DBeQ）被鉴定为一种选择性、有效、可逆和 ATP 竞争性的 p97 抑制剂。DBeQ 阻断了多种已通过 RNAi 证明依赖 p97 的过程，包括泛素融合降解和内质网相关降解途径报告物的降解，以及自噬体成熟。DBeQ 还能有效抑制癌细胞生长，并且比蛋白酶体抑制剂更能迅速动员效应半胱氨酸蛋白酶-3 和 -7。我们的研究结果为癌症治疗中靶向 p97 提供了依据。
