Methyltransferase Drug Discovery

描述（由申请人提供）：染色质组装和结构在DNA复制，基因表达和细胞周期的进展过程中受到高度调节。包括甲基化状态在内的表观遗传因素的功能障碍与多种癌症有关。组蛋白甲基转移酶（hmt），包括赖氨酸和精氨酸甲基转移酶，被认为是一类新的重要的药物靶点，然而高通量筛选（HTS）检测格式不可用，参考抑制剂仍然未知。建立HTS分析方法和参比化合物将有助于阐明这些酶的功能，并促进抗癌药物的开发。反应生物学公司（RBC）已经开发了许多酶类的极低成本反应系统，以服务于HTS药物发现，大规模IC50测定和选择性/毒性分析市场。RBC的HotSpot平台采用了超低体积放射性同位素分析激酶谱的金标准。这个激酶分析的服务产品在激酶抑制剂开发的药物发现界被广泛接受。基于这些技术，RBC正在开发一种新的检测平台，使用金标准放射性同位素检测组蛋白甲基转移酶（hmt）大家族。在第一阶段，RBC已经成功开发了超过13种甲基转移酶，使用我们的低成本放射性同位素为基础的格式。这些检测方法已经过内部和客户的验证。我们通过使用其中一种酶进行了HTS试验，并确定了一些泛甲基转移酶抑制剂，这些抑制剂正在进行大型小组分析和基于细胞的分析的进一步评估。最终，这些化合物可以作为表观遗传学研究的研究工具，这是缺乏参考化合物的。在第二阶段，我们建议扩展该技术以添加额外的20个甲基转移酶（目标1）。在目标2中，HTS活动使用45,000种不同结构的化合物库来对抗甲基转移酶的RBC panel，将建立一个数据库来驱动结构-活性关系（SAR）模型。在Aim 3中，所有的个体命中将被描述为体外选择性和效力，具有良好活性和干净结构的候选物将在基于细胞的甲基转移酶抑制试验中进一步评估，先导化合物将通过进一步的SAR研究。通过II期资金，RBC将能够提供HTS和分析服务，以覆盖大多数人类甲基转移酶，并为研究活动提供工具化合物。在这笔资金结束时，RBC将成为提供表观遗传药物发现所需的大部分工具的主要推动力。在II期之后，RBC将寻找潜在的合作来进一步研究我们在II期筛选过程中发现的抑制剂。由于这笔资金，药物发现实验室将有机会获得目前不存在的新型工业级筛选和分析分析，该领域的蛋白质生产也将得到升级。

DESCRIPTION (provided by applicant): Chromatin assembly and structure is highly regulated during DNA replication, gene expression, and progression through the cell cycle. Dysfunctions of epigenetic factors, including methylation states, are associated with a variety of cancers. Histone methyltransferases (HMTs), including lysine and arginine methyltransferases, are considered a new and important class of drug targets, however high throughput screening (HTS) assay formats are not available and reference inhibitors remain unknown. Establishment of HTS assays and reference compounds would have significant potential to help elucidating the function of these enzymes and to facilitate the development of anticancer agents. Reaction Biology Corporation (RBC) has developed extremely low cost reaction systems for many enzyme classes to serve markets for HTS drug discovery, large scale IC50 determinations and selectivity/toxicity profiling. RBC's HotSpot platform employs a gold standard for ultralow volume radioisotope assays for kinase profiling. This service product for kinase profiling is widely accepted in the drug discovery community for kinase inhibitor development. Based on these skills, RBC is developing a new assay platform using gold standard radioisotope assays for the large family of histone methyltransferases (HMTs). During Phase I, RBC has successfully developed over 13 methyltransferases by using our low cost radioisotope based format. These assays have been validated internally and by customers. We have conducted a trial HTS by using one of the enzymes and identified a few pan-methyltransferase inhibitors that are under further evaluation in both large panel profiling and in cell based assays. Eventually, these compounds could be used as research tools for epigenetic research, which is lacking reference compounds. In Phase II, we propose to expand this technology to add an additional 20 methyltransferases (Aim 1). In Aim 2, HTS campaigns using a 45,000 compound library of diverse structures against the RBC panel of methyltransferase will establish a database to drive structure-activity relationship (SAR) models. In Aim 3, all the individual hits will be profiled for in vitro selectivity and potency, candidates with good activity and clean structures will be further evaluated in cell based assays for methyltransferase inhibition, and lead compounds will go through further SAR studies. Through Phase II funding, RBC will be able to provide HTS and profiling service to cover majority of the human methyltransferases and providing tool compound for research activities. By the end of this funding, RBC will be the major driving force for providing most of the tools needed for epigenetic drug discoveries. After Phase II, RBC will looking for potential collaborations to further SAR studies on the inhibitors that we have discovered in the Phase II screening process. As a result of this funding, drug discovery labs will have access to a new class of industrial grade screening and profiling assays that do not exist today, and protein production in this area will be upgraded as well. PUBLIC HEALTH RELEVANCE: Chromatin assembly and structure is highly regulated during DNA replication, gene expression, and progression through the cell cycle. Dysfunctions of epigenetic factors, including methylation states, are associated with a variety of cancers. Histone methyltransferases (HMTs), including lysine and arginine methyltransferases, are considered a new and important class of drug targets, however high throughput screening (HTS) assay formats are not available and reference inhibitors remain unknown. Establishment of HTS assays and reference compounds would have significant potential to help elucidating the function of these enzymes and to facilitate the development of anticancer agents. Reaction Biology Corporation (RBC) has developed extremely low cost reaction systems for many enzyme classes to serve markets for HTS drug discovery, large scale IC50 determinations and selectivity/toxicity profiling. The HotSpot platform employs a gold standard for ultralow volume radioisotope assays for kinase profiling. This service product for kinase profiling is widely accepted in the drug discovery community for kinase inhibitor development. Based on these skills, RBC seeks to develop a new assay platform using gold standard radioisotope assays for the large family of histone methyltransferases.