Methyltransferase Drug Discovery

描述(由申请人提供)：染色质的组装和结构在DNA复制、基因表达和细胞周期的进展过程中受到高度调控。表观遗传因素的功能障碍，包括甲基化状态，与多种癌症有关。组蛋白赖氨酸甲基转移酶(HKMT)被认为是一类新的和重要的药物靶点，然而高通量筛选(HTS)分析模式还不可用，参考抑制剂仍然未知。HTS分析和参考化合物将在帮助阐明这些酶的功能和促进抗癌药物的开发方面具有重要的潜力。反应生物学公司(RBC)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(UHTS)药物发现、大规模IC50测定和选择性/毒性分析市场。HotSpot平台采用了超低容量放射性同位素分析的黄金标准来进行激酶分析。这一用于激酶分析的服务产品在用于开发激酶抑制剂的药物发现社区中被广泛接受。基于这些技能，RBC寻求开发一种新的分析平台，使用组蛋白赖氨酸甲基转移酶(HKMT)大家族的金标准放射性同位素分析。透过第一阶段的拨款，拟议的研究将集中于以下三个目标：目标1发展一个以放射性同位素为基础的通用金标准，以供大讯号/背景比的高温超导之用；目标2为强健高温超导的超小型化和简化化验程序，以降低化验成本；目标3进行高温超导测试，并透过从红细胞化学资料库中筛选文库，利用目标1和2所发展的化验方法，为香港MT鉴定新化合物。这项工作将使第二阶段的活动成为可能，在这一阶段，红细胞将扩大其在大型人类HKMT家族中的检测范围，并使用这一新开发的针对大型合成和天然化合物库的通用检测来发现用于研究和治疗开发的潜在分子。选定的先导化合物将在基于细胞的分析中进行抗癌活动的公共卫生相关性评估：染色质组装和结构在DNA复制、基因表达和细胞周期中的进展过程中受到高度调控。表观遗传因素的功能障碍，包括甲基化状态，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和重要的药物靶点，然而高通量筛选分析模式是可用的，参考抑制剂仍然未知。HTS分析和参考化合物将在帮助阐明这些酶的功能和促进抗癌药物的开发方面具有重要的潜力。反应生物公司已经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平台采用了超低容量放射性同位素分析的黄金标准来进行激酶分析。这一用于激酶分析的服务产品在用于开发激酶抑制剂的药物发现社区中被广泛接受。基于这些技能，RBC寻求开发一种新的分析平台，使用组蛋白赖氨酸甲基转移酶大家族的金标准放射性同位素分析。

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.