A NOVEL APPROACH TO THE DISCOVERY OF ANTIBACTERIAL DRUGS

对主要类别抗生素产生抗药性的细菌病原体的出现对公共健康构成了严重威胁，并产生了对新型抗菌剂的需求。这些新的抗菌剂应该针对新的药物靶点发挥作用。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是，对于许多这些靶点来说，对它们的生化功能知之甚少或一无所知。这笔赠款建议开发一个平台，以识别作用于未知功能目标的新型抗菌剂。初步结果表明，用噬菌体展示技术分离得到的酶特异性多肽可以作为检测酶抑制剂的探针。这项拨款的研究将涉及从大肠杆菌中克隆、过度生产和提纯三种必需的基因产品。然后，将使用噬菌体展示技术识别与这些必需基因产物中的每一种特异结合的多肽。通过在大肠杆菌中表达多肽并监测细胞生长，可以验证多肽对必要功能的抑制作用。验证过的多肽将用于形成分析并进行高通量筛选。HTS的点击将进行抗菌活性测试。拟议的商业应用多肽配体将在高通量筛选分析中用作分子探针，以识别抗菌药物开发中的小化合物(先导)。这项技术将提供一种方法来识别生化活性未知的靶标的抑制剂。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上。

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.