title: Synthesis and structure-activity relationship studies of N-benzyl-2-phenylpyrimidin-4-amine derivatives as potent USP1/UAF1 deubiquitinase inhibitors with anticancer activity against nonsmall cell lung cancer.
abstract: Deregulation of ubiquitin conjugation or deconjugation has been implicated in the pathogenesis of many human diseases including cancer. The deubiquitinating enzyme USP1 (ubiquitin-specific protease 1), in association with UAF1 (USP1-associated factor 1), is a known regulator of DNA damage response and has been shown as a promising anticancer target. To further evaluate USP1/UAF1 as a therapeutic target, we conducted a quantitative high throughput screen of >400000 compounds and subsequent medicinal chemistry optimization of small molecules that inhibit the deubiquitinating activity of USP1/UAF1. Ultimately, these efforts led to the identification of ML323 (70) and related N-benzyl-2-phenylpyrimidin-4-amine derivatives, which possess nanomolar USP1/UAF1 inhibitory potency. Moreover, we demonstrate a strong correlation between compound IC50 values for USP1/UAF1 inhibition and activity in nonsmall cell lung cancer cells, specifically increased monoubiquitinated PCNA (Ub-PCNA) levels and decreased cell survival. Our results establish the druggability of the USP1/UAF1 deubiquitinase complex and its potential as a molecular target for anticancer therapies.
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health , 9800 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Dexheimer
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Rosenthal
authors.forename: Diane K
authors.initials: DK
authors.lastname: Luci
authors.forename: Qin
authors.initials: Q
authors.lastname: Liang
authors.forename: Mark A
authors.initials: MA
authors.lastname: Villamil
authors.forename: Junjun
authors.initials: J
authors.lastname: Chen
authors.forename: Hongmao
authors.initials: H
authors.lastname: Sun
authors.forename: Edward H
authors.initials: EH
authors.lastname: Kerns
authors.forename: Anton
authors.initials: A
authors.lastname: Simeonov
authors.forename: Ajit
authors.initials: A
authors.lastname: Jadhav
authors.forename: Zhihao
authors.initials: Z
authors.lastname: Zhuang
authors.forename: David J
authors.initials: DJ
authors.lastname: Maloney
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D029681:Arabidopsis Proteins / Q000037:antagonists & inhibitors*; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy*; D045744:Cell Line, Tumor; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011743:Pyrimidines / Q000138:chemical synthesis* / Q000494:pharmacology; D013329:Structure-Activity Relationship; D064570:Ubiquitin-Specific Proteases / Q000037:antagonists & inhibitors*; D054875:Ubiquitination
chemical_list: D000970:Antineoplastic Agents; D029681:Arabidopsis Proteins; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D011743:Pyrimidines; D064570:Ubiquitin-Specific Proteases; C526165:WDR48 protein, human; C579228:USP1 protein, human