A NOVEL APPROACH TO THE DISCOVERY OF ANTIBACTERIAL DRUGS

对主要类别的抗生素具有耐药性的细菌病原体的出现构成了严重的公共卫生威胁，并产生了对新型抗菌剂的需求。 这些新的抗菌药物应直接作用于新的药物靶点。 细菌基因组学将提供许多新的潜在药物靶点。 然而，抗菌药物发现的局限性在于，对于这些靶点中的许多靶点，很少或根本不知道它们的生化功能。 该资助计划开发一个平台，以识别作用于未知功能靶标的新型抗菌剂。 初步结果表明，通过噬菌体展示分离的酶特异性肽可以用作探针来检测酶抑制剂。这项研究将涉及从大肠杆菌中克隆、过量生产和纯化三种必需的基因产物。杆菌然后使用噬菌体展示来鉴定与这些必需基因产物中的每一种特异性结合的肽。通过在E.大肠杆菌，并监测细胞生长。 经验证的肽将用于形成测定并进行高通量筛选。 将测试来自HTS的命中物的抗菌活性。肽配体将在高通量筛选试验中用作分子探针，以鉴定抗菌药物发现中的小化合物（先导化合物）。 这项技术将提供一种方法，以确定抑制剂的目标，其生化活性是未知的。 我们预计，从我们的筛选策略中获得的一些化合物线索将转移到药物开发中。

The emergence of bacterial pathogens that are resistant to major classes of antibiotics poses a serious public health threat and has created demand for novel antibacterial agents. These new antibacterial agents should be directed to act on novel drug targets. Bacterial genomics will provide many new potential drug targets. The limitation for antibacterial drug discovery, however, is that for many of these targets there will be little or no knowledge of their biochemical function. This grant proposes to develop a platform to identify novel antibacterial agents that act on targets of unknown function. Preliminary results have shown that enzyme-specific peptides isolated by phage display can be used as probes to detect enzyme inhibitors. The research in this grant would involve cloning, overproducing, and purifying three essential gene products from E. coli. Peptides would then be identified, using phage display, that specifically bound to each of these essential gene products. Inhibition of an essential function by the peptides would be validated by expressing the peptides in E. coli and monitoring cell growth. Validated peptides would be used to format an assay and perform a high throughput screen. Hits from the HTS would be tested for antibacterial activity. PROPOSED COMMERCIAL APPLICATION Peptide ligands will be used as molecular probes in high throughput screening assays to identify small chemical compounds (leads) in antibacterial drug discovery. This technology will provide a method to identify inhibitors of targets whose biochemical activity is unknown. We anticipate that some of the compound leads obtained from our screening strategy will move on to drug development.