Development of a dual screening assay for the efficient search for antiviral Hepatitis C entryinhibitors from microorganisms

寻找更有效的抗病毒物质，减少副作用是药物研究的主要课题。目前，只有少数病毒感染性疾病有令人满意的药物治疗。天然产物库仍然是新候选药物和先导结构的极好来源。尽管天然产物具有巨大的潜力，但由于基于细胞的抗病毒测定的复杂性和缺乏高通量，很少测试其抗病毒活性。我的研究项目旨在开发一种双重筛选方法，可以更有效地筛选天然产物库，并以中等通量筛选抗病毒活性。为此，生物传感器平台Octet® RED 96用于在新开发的基于蛋白质的结合测定中鉴定与靶蛋白具有高亲和力的小分子。在这个项目中，我将使用丙型肝炎病毒（HCV）包膜蛋白E1/E2来寻找潜在的新的进入抑制剂。然后在基于细胞的试验中，使用HCV假型病毒，通过单轮HCV感染性试验评价该结合试验的命中物的进入抑制活性。活性化合物将在结构、活性以及与病毒蛋白的结合方面得到充分表征，并且它们将立即作为有价值的先导结构。这种双重筛选概念提供了一种更有效的筛选用于药物治疗的天然产物的方法。这两种体外试验的结合有可能加速药物发现过程，并发现抗病毒化合物和结构线索，这将有助于开发新的抗病毒药物或作为研究病毒感染的分子探针。

The search for more efficient antiviral substances with less side effects is a predominant topic in drug research. Currently, only few viral infectious diseases have satisfactory drug therapies. The pool of natural products continues to be an excellent source for new drug candidates and lead structures. Despite of their great potential natural products are very rarely tested for antiviral activity due to the complexity of cell-based antiviral assays and their lack of high throughput. My research project aims to develop a dual screening approach that allows to screen natural product libraries more efficiently and in a medium throughput for antiviral activity. For this purpose, the biosensor platform Octet® RED96 is used to identify small molecules with high affinities to a target protein in a newly developed protein-based binding assay. For this project I will use the Hepatitis C (HCV) envelope protein E1/E2 to find potential new entryinhibitors. Hits from this binding assay will then be evaluated for entryinhibitory activity in a cell-based assay with single round HCV infectivity assay using HCV-pseudotyped viruses. Active compounds will be fully characterized in terms of structure, activity as well as binding to the viral protein and they will serve immediately as valuable lead structures.This dual screening concept offers a substantially more efficient way to screen for natural products for drug therapy. The combination of the two in vitro assays has the potential to accelerate the drug discovery process and to find antiviral compounds and structural leads that will contribute to the development of new antiviral drugs or serve as molecular probes to study viral infections.