Diaz Elsie, Machutta Carl A, Chen Stephanie, Jiang Yong, Nixon Christopher, Hofmann Glenn, Key Danielle, Sweitzer Sharon, Patel Mehul, Wu Zining, Creasy Caretha L, Kruger Ryan G, LaFrance Louis, Verma Sharad K, Pappalardi Melissa B, Le Baochau, Van Aller Glenn S, McCabe Michael T, Tummino Peter J, Pope Andrew J, Thrall Sara H, Schwartz Benjamin, Brandt Martin
Platform Technology and Science, GlaxoSmithKline, Collegeville, PA 19426, USA.
J Biomol Screen. 2012 Dec;17(10):1279-92. doi: 10.1177/1087057112453765. Epub 2012 Aug 17.
Histone methyltransferases (HMT) catalyze the methylation of histone tail lysines, resulting in changes in gene transcription. Misregulation of these enzymes has been associated with various forms of cancer, making this target class a potential new area for the development of novel chemotherapeutics. EZH2 is the catalytic component of the polycomb group repressive complex (PRC2), which selectively methylates histone H3 lysine 27 (H3K27). EZH2 is overexpressed in prostate, breast, bladder, brain, and other tumor types and is recognized as a molecular marker for cancer progression and aggressiveness. Several new reagents and assays were developed to aid in the identification of EZH2 inhibitors, and these were used to execute two high-throughput screening campaigns. Activity assays using either an H3K27 peptide or nucleosomes as substrates for methylation are described. The strategy to screen EZH2 with either a surrogate peptide or a natural substrate led to the identification of the same tractable series. Compounds from this series are reversible, are [(3)H]-S-adenosyl-L-methionine competitive, and display biochemical inhibition of H3K27 methylation.
组蛋白甲基转移酶（HMT）催化组蛋白尾部赖氨酸的甲基化，从而导致基因转录发生变化。这些酶的失调与多种癌症相关，使得该靶点类别成为新型化疗药物开发的一个潜在新领域。EZH2是多梳蛋白抑制复合物（PRC2）的催化成分，它能选择性地使组蛋白H3赖氨酸27（H3K27）甲基化。EZH2在前列腺癌、乳腺癌、膀胱癌、脑癌和其他肿瘤类型中过度表达，被认为是癌症进展和侵袭性的分子标志物。开发了几种新的试剂和检测方法以辅助EZH2抑制剂的鉴定，并将其用于开展两项高通量筛选活动。描述了使用H3K27肽或核小体作为甲基化底物的活性检测方法。用替代肽或天然底物筛选EZH2的策略导致鉴定出相同的易处理系列。该系列化合物是可逆的，具有[（3）H]-S-腺苷-L-甲硫氨酸竞争性，并且对H3K27甲基化具有生化抑制作用。
