Epigenetics Probes: Production of histone modifying enzymes and identification o

描述(由申请人提供)：组蛋白修饰酶的生产和化学探针的鉴定用于表观遗传学发现基因组功能是最常见疾病的主要因素。基因组功能受组蛋白翻译后甲基化和乙酰化状态的调节。组蛋白是在细胞核中发现的一种蛋白质，它能产生核小体，DNA就是其中的结构。修饰组蛋白的酶是癌症、神经退行性疾病、心血管疾病和代谢性疾病等适应症的重要潜在药物靶点。催化这些修饰的酶包括许多组蛋白甲基转移酶和去甲基酶(MT和DMT)。此外，识别或“阅读”这些组蛋白修饰的蛋白质含有溴域、色域等，也被认为是表观遗传因子。如果药物要超越仅代表对症治疗的传统药物靶点，那么验证这些表观遗传酶和因子作为药物靶点仍然是药物发现的首要任务。化学表观遗传抑制剂在这里代表着一个强大的机会。然而，组蛋白甲基化是一个相对较新的发现空间，MT或DMT抑制剂的可能结构尚不清楚。无论是用于实验室研究还是用于铅开发，几乎没有小分子抑制剂可用。因此，迫切需要为这些靶标开发分析方法和确定新的化学探针。小分子探针将在科学研究中发挥有价值的作用，并作为铅开发的药物化学支架。在这项提案中，我们将采用广域高通量筛选(HTS)方法，测试1400多种特权支架，其中包括大多数FDA批准的药物和临床试验试剂，以对抗30多种组蛋白修饰酶，包括HMT和组蛋白去甲基酶(HDMT)。我们的初步研究已经显示了非常积极的结果，确定并确认了对少数选定的HMT的命中。已知的口服活性和相对安全的化合物的广域高温超导为化学探针的发现提供了一些优势：(1)任何新发现的活性化合物都将具有支持新应用的已知科学药理学；(2)这些探针有可能实现“药物拯救”或“药物再利用”；(3)对于已经上市的化合物，新的表观遗传活性信息将有助于在没有数据的表观遗传水平上加强对其有效性或毒性的了解；(4)对于目前正在进行临床试验或未能通过后期临床试验的化合物，新的信息将为修改这些化合物以获得更好的定向治疗效果提供指导，以及(5)这些数据将有助于建立一个通用的化学表观基因组数据库。拥有一个公开可用的表观遗传抑制物库对于在这一领域取得进展是至关重要的。

DESCRIPTION (provided by applicant): Production of histone modifying enzymes and identification of chemical probes for epigenetic discovery Genomic function is the major factor in the most common diseases. Genomic function is regulated by the posttranslational methylation and acetylation state of histones. Histones are proteins found in cell nuclei that create nucleosomes into which DNA is structured. Enzymes that modify histones represent important potential drug targets for indications like cancers, neurodegenerative disorders, cardiovascular, and metabolic diseases. Enzymes that catalyze these modifications include numerous histone methyltransferases and demethylases ("MT's" & "DMT's"). Additionally, proteins that recognize or "read" these histone modifications contain bromodomains, chromodomains, etc. and are also considered epigenetic factors. If medicine is to move beyond conventional drug targets that represent symptomatic treatment only, then validation of these epigenetic enzymes and factors as drug targets remains a top priority for drug discovery. Chemical epigenetic inhibitors represent a powerful opportunity here. However, histone methylation is a relatively new discovery space, and the likely structures of MT or DMT inhibitors are not yet known. Few small molecule inhibitors are available, either for laboratory research or for lead development. Therefore, there is a significant and urgent need to develop assays and to identify new chemical probes for these targets. Small molecule probes would serve a valuable role in scientific investigations and serve as medicinal chemistry scaffolds for lead development. In this proposal, we will deploy a wide-field high throughput screening (HTS) approach by testing more than 1400 privileged scaffolds that include most FDA approved drugs and clinical trial agents against over 30 histone modifying enzymes, including HMTs and histone demethylases (HDMTs). Our preliminary studies have already demonstrated very positive results by identifying and confirming hits against a handful of selected HMTs. Wide-field HTS with known orally active and relatively safe compounds offers a number of advantages for chemical probe discovery: (1) any newly identified active compound will have a known scientific pharmacology to support new applications; (2) these probes have the potential to enable "drug rescue" or "drug repurposing"; (3) for compounds already on the market, the new epigenetic activity information will help to provide enhanced understanding of their effectiveness or toxicity at the epigenetic level where no data was available; (4) for the compounds that are currently in clinical trials or have failed i late stage clinical trials, the new information will provide guidance to modify these compounds for better directed therapeutic effectiveness, and (5) this data will help establish a general chemo-epigenomic database. Having a publicly available library of epigenetic inhibitors is essential to moving ahead in this area.