Characterisation of the antiviral properties and mechanism of action of natural product MM 46115

甲型流感病毒是严重呼吸道感染的主要原因，在英国每年导致2万人住院。针对该病毒的疫苗已经开发出来，以防止严重感染，但疫苗的犹豫不决限制了它们的用途。给住院患者服用抗病毒药物，以防止病毒复制。然而，病毒已经成功地对这些药物产生了抗药性，使它们无效。因此，迫切需要新的有效治疗方法。据报道，引起真菌瘤的放线菌能产生MM 46115，这是一种螺四磷酸聚酮，对多种病毒有活性，包括甲型流感。MM 46115可能有潜力被开发成一种新的治疗方法，克服目前使用的抗病毒药物的耐药性。先前的工作表明，用MM 46115处理流感感染细胞可以减少细胞死亡，而不会产生毒副作用。虽然MM 46115的平面结构已经确定，但它的相对和绝对立体化学仍有待充分阐明。这是为了更好地了解其与其他螺四磷酸多酮的结构相似性，并开始阐明其构效关系。本项目旨在通过空斑分析、RT-PCR法和免疫荧光法进一步研究MM 46115对甲型流感的抗病毒活性。MM 46115的立体化学将结合生物合成酶的预测序列分析、2D核磁共振光谱、圆二色谱、X射线结晶学和微电子衍射来阐明。蛋白质组学还将用于确定MM 46115的蛋白质靶标。最后，将更好地了解MM46115的生物合成，使生物工程方法能够生产提供结构-活性关系的结构类似物。

The influenza A virus is a major cause of severe respiratory infection, which results in 20,000 hospitalisations per year in the UK. Vaccines have been developed against the virus to prevent severe infections, but vaccine hesitancy limits their utility. Antiviral drugs are administered to hospitalised patients to prevent replication of the virus. However, the virus has managed to evolve resistance to these drugs, rendering them ineffective. Therefore, there is a pressing need for new and effective treatments. Actinomadura pelletieri, a bacterium that causes mycetoma, has been reported to produce MM 46115, a spirotetronate polyketide active against multiple viruses, including influenza A. MM 46115 may have the potential to be developed into a novel treatment that overcomes resistance to currently used antiviral drugs. Previous work has shown that treating influenza infected cells with MM 46115 can reduce cell death without toxic side effects. While the planar structure of MM 46115 has been determined, its relative and absolute stereochemistry remains to be fully elucidated. This is required to develop a better understanding of its structural similarity to other spirotetronate polyketides and begin illuminating the structure-activity relationship.This project aims to further investigate the antiviral activity of MM 46115 against influenza A using plaque assays, RT-PCR and immunofluorescence. The stereochemistry of MM 46115 will be elucidated using a combination of predictive sequence analysis of biosynthetic enzymes, 2D NMR spectroscopy, CD spectroscopy, X-ray crystallography, and micro electron diffraction. Proteomics will also be used to identify protein targets for MM 46115. Finally, a better understanding of MM46115 biosynthesis will be developed, enabling bioengineering approaches to the production of structural analogues that inform the structure-activity relationship to be employed.