Substrate Profiling of New Enzymes for Drug Discovery

用于药物发现的新酶的底物分析

• 批准号: 6403853
• 负责人: GREGORY P CONNELLY
• 金额: $ 52.7万
• 依托单位: ALTHEXIS COMPANY, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-15 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6403853
• 关键词: Escherichia coli; antibacterial agents; bacterial genetics; bacterial proteins; bioassay; chemical registry /resource; chemical synthesis; crystallization; drug design /synthesis /production; enzyme induction /repression; enzyme inhibitors; enzyme substrate; enzyme substrate analog; functional /structural genomics; high performance liquid chromatography; high throughput technology; intermolecular interaction; ligands; microcalorimetry; molecular cloning; polymerase chain reaction; protein purification; protein structure function; technology /technique development

DESCRIPTION (provided by applicant): With a dramatic increase in the number of complete bacterial genomes in the public and private databases, there exists an outstanding opportunity for the identification of new antibacterial protein targets. Interestingly, the accumulation of genomic sequences has been punctuated by the realization that typically greater than one third of any genome cannot be annotated for function based on sequence similarity. Several of these "unknowns" have been shown to be genetically validated drug discovery targets. However, without a defined biochemical activity, unknowns that prove to be essential for growth and viability are intractable for the purposes of drug discovery. Biochemical assay of protein function is critical to the implementation of high-throughput screening to discover lead compounds, to the determination of a detailed enzyme mechanism, and to the successful optimization of those leads to tight-binding inhibitors and ultimately to efficacious drugs. The proposed research aims to break new ground in the problem of defining general functional assays, and in understanding the biochemical activities of proteins of unknown function. The experimental basis for our proposed methodology involves microcalorimetry and substrate libraries. In this research proposal, we aim to demonstrate the technology by identifying substrates for enzymes of unknown function from E. coli, determining the enzyme mechanisms of these proteins, and designing inhibitors based on the mechanisms. Establishment of the technology proposed here enables a procedure for the selection of validated enzyme targets and for the identification of novel antibacterial drug candidates to combat drug resistant pathogens. PROPOSED COMMERCIAL APPLICATION: The invention disclosed here provides a method for identifying substrates and inhibitors for new enzyme targets identified through genomic sequencing. As applied to proteins of unknown function from bacterial genomes, the method of substrate profiling allows one to establish assays for anti-bacterial lead compound discovery in the pharmaceutical industry. The annual world wide market for antibiotic compounds is in excess of $23 billion. Single product sales can exceeed $1 billion. The technology described here enables the discovery of novel antibiotics.

描述（由申请人提供）：数量急剧增加 在公共和私人数据库中完整的细菌基因组，存在 鉴定新抗菌蛋白的出色机会 目标。有趣的是，基因组序列的积累已经 被认识到通常大于任何人的三分之一的认识所打断 基于序列相似性的功能不能注释基因组。一些 这些“未知数”已被证明是经过遗传验证的药物发现 目标。但是，没有定义的生化活动，未知的证明 对于生长和生存能力至关重要 药物发现。蛋白质功能的生化测定对于 实施高通量筛选以发现铅化合物， 确定详细的酶机制，并成功地确定 优化这些导致紧密结合抑制剂，最终导致 有效的药物。拟议的研究旨在在 定义一般功能测定和理解的问题 功能未知的蛋白质的生化活性。实验基础 对于我们提出的方法，涉及微钙化和底物库。 在这项研究建议中，我们旨在通过确定 大肠杆菌的未知功能酶的底物，确定酶 这些蛋白质的机制，并根据机制设计抑制剂。 这里提出的技术的建立可以为 选择经过验证的酶靶标和用于鉴定新颖的 抗菌药物候选者以对抗耐药病原体。 拟议的商业应用： 此处披露的本发明提供了一种方法，用于识别通过基因组测序鉴定的新酶靶标的底物和抑制剂。正如细菌基因组中未知功能的蛋白质所应用的，底物分析的方法使人们可以在制药行业中建立抗细菌铅复合发现的测定法。年度全球抗生素化合物市场超过230亿美元。单产品销售可以省去10亿美元。此处描述的技术可以发现新型抗生素。