Abstract LB-11: Discovery, synthesis, and structure-activity relationship of N-benzyl-2-phenylpyrimidin-4-amine derivatives as potent USP1/UAF1 deubiquitinase inhibitors with anticancer activity against non-small cell lung cancer

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Protein ubiquitination is a dynamic and reversible post-translational modification that has been linked to many essential cellular processes in eukaryotes. In addition, deregulation of ubiquitin conjugation or deconjugation has been implicated in the pathogenesis of many human diseases, including cancer. For example, the deubiquitinating enzyme USP1 (ubiquitin-specific protease 1) in association with its WD40-repeat protein binding partner, UAF1 (USP1-associated factor 1), is a known regulator of DNA damage tolerance and repair and has been proposed as a promising target for anticancer therapy. To further evaluate the USP1/UAF1 complex as a therapeutic target, we conducted a quantitative high throughput screen of >400,000 compounds and subsequent medicinal chemistry optimization in pursuit of small molecules that inhibit the deubiquitinating activity of USP1/UAF1. These efforts lead to the identification of ML323 and a series of N-benzyl-2-phenylpyrimidin-4-amine derivatives, which possess nanomolar USP1/UAF1 inhibition and exhibit excellent selectivity over related proteases. Moreover, we demonstrate a strong correlation between compound IC50 values for USP1/UAF1 inhibition in vitro and activity in non-small cell lung cancer cells, specifically increasing the level of ubiquitinated-PCNA and decreasing cell survival. Taken together, our results establish the druggability of the USP1/UAF1 deubiquitinase complex and its potential as molecular target for anticancer therapies. Citation Format: Thomas S. Dexheimer, Andrew S. Rosenthal, Diane Luci, Qin Liang, Mark A. Villamil, Ajit Jadhav, Anton Simeonov, Zhihao Zhuang, David J. Maloney. Discovery, synthesis, and structure-activity relationship of N-benzyl-2-phenylpyrimidin-4-amine derivatives as potent USP1/UAF1 deubiquitinase inhibitors with anticancer activity against non-small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-11. doi:10.1158/1538-7445.AM2014-LB-11