An Orally Bioavailable Chemical Probe of the Lysine Methyltransferases EZH2 and EZH1

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• 作者：Kyle D. Konze ; Anqi Ma ; Fengling Li ; Dalia Barsyte-Lovejoy ; Trevor Parton ; Christopher J. MacNevin ; Feng Liu ; Cen Gao ; Xi-Ping Huang ; Ekaterina Kuznetsova ; Marie Rougie ; Alice Jiang ; Samantha G. Pattenden ; Jacqueline L. Norris ; Lindsey I. James ; Bryan L. Roth ; Peter J. Brown ; Stephen V. Frye ; Cheryl H. Arrowsmith ; Klaus M. Hahn ; Gang Greg Wang ; Masoud Vedadi ; Jian Jin
• 刊名：ACS Chemical Biology
• 出版年：2013
• 出版时间：June 21, 2013
• 年：2013
• 卷：8
• 期：6
• 页码：1324-1334
• 全文大小：614K
• 年卷期：v.8,no.6(June 21, 2013)
• ISSN：1554-8937

文摘

EZH2 or EZH1 is the catalytic subunit of the polycomb repressive complex 2 that catalyzes methylation of histone H3 lysine 27 (H3K27). The trimethylation of H3K27 (H3K27me3) is a transcriptionally repressive post-translational modification. Overexpression of EZH2 and hypertrimethylation of H3K27 have been implicated in a number of cancers. Several selective inhibitors of EZH2 have been reported recently. Herein we disclose UNC1999, the first orally bioavailable inhibitor that has high in vitro potency for wild-type and mutant EZH2 as well as EZH1, a closely related H3K27 methyltransferase that shares 96% sequence identity with EZH2 in their respective catalytic domains. UNC1999 was highly selective for EZH2 and EZH1 over a broad range of epigenetic and non-epigenetic targets, competitive with the cofactor SAM and non-competitive with the peptide substrate. This inhibitor potently reduced H3K27me3 levels in cells and selectively killed diffused large B cell lymphoma cell lines harboring the EZH2^Y641N mutant. Importantly, UNC1999 was orally bioavailable in mice, making this inhibitor a valuable tool for investigating the role of EZH2 and EZH1 in chronic animal studies. We also designed and synthesized UNC2400, a close analogue of UNC1999 with potency >1,000-fold lower than that of UNC1999 as a negative control for cell-based studies. Finally, we created a biotin-tagged UNC1999 (UNC2399), which enriched EZH2 in pull-down studies, and a UNC1999鈥揹ye conjugate (UNC2239) for co-localization studies with EZH2 in live cells. Taken together, these compounds represent a set of useful tools for the biomedical community to investigate the role of EZH2 and EZH1 in health and disease.