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。组蛋白赖氨酸甲基转移酶（HKMTs）被认为是一类新的重要药物靶点，但高通量筛选（HTS）检测格式不可用，参考抑制剂仍然未知。HTS分析和参比化合物将有很大的潜力帮助阐明这些酶的功能，并促进抗癌药物的开发。反应生物学公司（RBC）开发了许多酶类的极低成本反应系统，以服务于超高通量筛选（uHTS）药物发现，大规模IC50测定和选择性/毒性分析市场。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寻求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赖氨酸甲基转移酶（HKMTs）大家族。在第一阶段的资助下，建议的研究将集中于以下三个目标：目标一为信号/背景比大的HKMTs制定通用的放射性同位素金标准；目的2：通过超小型化和简化稳健HTS的检测方案来降低检测成本；目标3：通过筛选RBC化学库中的文库，使用目标1和2中开发的分析方法进行HTS和鉴定HKMT的新化合物。这项工作将使二期活动成为可能，届时RBC将扩大其在大型人类HKMTs家族中的检测范围，并使用这种新开发的通用检测方法针对大型合成和天然化合物文库，发现潜在的分子用于研究和治疗开发。选定的先导化合物将在基于细胞的检测中评估其抗癌活性。公共卫生相关性：在DNA复制、基因表达和细胞周期进展过程中，染色质组装和结构受到高度调控。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。组蛋白赖氨酸甲基转移酶被认为是一类新的重要的药物靶点，然而高通量筛选分析格式不可用，参考抑制剂仍然未知。HTS分析和参比化合物将有很大的潜力帮助阐明这些酶的功能，并促进抗癌药物的开发。Reaction Biology Corporation为许多酶类开发了极低成本的反应系统，以服务于超高通量筛选药物发现，大规模IC50测定和选择性/毒性分析的市场。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。基于这些技能，RBC寻求开发一种新的检测平台，使用金标准放射性同位素检测大家族的组蛋白赖氨酸甲基转移酶。

DESCRIPTION (provided by applicant): Chromatin assembly and structure is highly regulated during DNA replication, gene expression, and progression through the cell cycle. Dysfunction of epigenetic factors, including methylation state, are associated with a variety of cancers. Histone lysine methyltransferases (HKMTs) 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. HTS assays and reference compounds would have significant potential to help elucidate 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 ultra high throughput screening (uHTS) 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 lysine methyltransferases (HKMTs). Through Phase I funding, the proposed research will focus on the following three aims: Aim 1 Develop a universal radioisotope-based gold standard for HKMTs with large signal/background ratio; Aim 2 Reduce assay costs by ultra-miniaturization and streamline assay protocols for robust HTS; Aim 3 Conduct HTS and identify new compounds for an HKMT using assays developed in Aims 1 and 2 by screening the libraries from RBC's chemical repository. This work will enable Phase II activities where RBC will expand its assay range across the large human HKMTs family and use this newly developed universal assay against large synthetic and natural compound libraries to discover potential molecules for research and therapeutic development. Selected lead compounds will be evaluated in cell based assays for anti-cancer activities PUBLIC HEALTH RELEVANCE: Chromatin assembly and structure is highly regulated during DNA replication, gene expression, and progression through the cell cycle. Dysfunction of epigenetic factors, including methylation state, are associated with a variety of cancers. Histone lysine methyltransferases are considered a new and important class of drug targets, however high throughput screening assay formats are not available and reference inhibitors remain unknown. HTS assays and reference compounds would have significant potential to help elucidate function of these enzymes and to facilitate the development of anticancer agents. Reaction Biology Corporation has developed extremely low cost reaction systems for many enzyme classes to serve markets for ultra high throughput screening 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 lysine methyltransferases.