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20，000人住院。已经开发了针对该病毒的疫苗来预防严重感染，但疫苗的犹豫限制了它们的效用。对住院患者给予抗病毒药物以防止病毒复制。然而，病毒已经设法进化出对这些药物的耐药性，使它们无效。因此，迫切需要新的有效治疗方法。据报道，引起足菌肿的Actinomadura pelletieri细菌可产生MM 46115，MM 46115是一种对多种病毒（包括甲型流感病毒）具有活性的螺内酯聚酮。MM 46115可能有潜力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。先前的研究表明，用MM 46115处理流感感染的细胞可以减少细胞死亡，而没有毒副作用。虽然MM 46115的平面结构已经确定，但其相对和绝对立体化学仍有待充分阐明。这需要更好地了解其结构相似性的其他spirotetronate聚ketides和开始阐明的结构-活性relationship.This项目旨在进一步研究MM 46115抗甲型流感病毒的抗病毒活性，使用空斑试验，RT-PCR和免疫荧光。将使用生物合成酶的预测序列分析、2D NMR光谱、CD光谱、X射线晶体学和微电子衍射的组合来阐明MM 46115的立体化学。蛋白质组学还将用于识别MM 46115的蛋白质靶点。最后，将开发MM 46115生物合成的更好的理解，使生物工程方法生产的结构类似物，通知的结构-活性关系被采用。

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.