流感病毒严重危害人类健康，病毒变异导致临床药物失效。发现新作用机制、高效低毒且不易耐药抗流感病毒天然药源分子迫在眉睫。海洋微生物具备抗病毒胁迫和化学防御功能，是抗流感病毒新型药源分子最有潜力的生物资源。课题基于前期从海洋微生物库中发现具有显著抑制流感病毒菌株及其抗病毒成分的研究基础，设计以病毒重要靶标为导向的谱效相关等方法，从分子库中高效获得对流感病毒进入和复制关键靶酶有显著靶向作用的活性分子，并进行抗病毒药效评价，获得新型抗流感病毒药源分子。基于抗流感病毒活性菌种的合成基因簇等生物信息，应用合成生物学等技术挖掘全新结构抗流感病毒分子。此外，对已明确抗流感病毒作用的天然分子进行深入抗病毒作用机制研究，阐明活性分子对关键靶蛋白的调控作用；并对其中具有抗临床耐药毒株且对病毒低耐药的化合物开展抗耐药作用分子机制研究，揭示抗耐药靶向因子。课题研究将为抗流感病毒药物研究提供全新药源分子。

Influenza virus induced serious harm to human health, while the virus mutation led to failure treatment for clinical drugs. Thus, it is imminent to discover new anti-influenza virus agents which possess strong efficiency and low toxicity as well as the action with new mechanistic pathway which may be used for the remedy of drug-resistant viral species. Marine derived microorganisms inhabited under ecological stresses and possessed chemical defense capabilities, they are thus considered as the new and promising source provided for the finding of new anti-influenza virus lead compounds. Based on our previous achievement to obtain a series of marine microorganisms with potential inhibition against influenza virus strains and handled several hundreds of marine microorganism-derived new natural products and the results of their antivirus assay, this project aimed to discover structurally novel natural products from marine microorganisms with potential anti-influenza virus activity which are able to be developed as lead compounds. A protocol is designed using viral enzyme-oriented UF-LC-MSn method and other related methods for efficiently obtaining active molecules from microorganism library toward the key enzymes (HA, DNA polymerase, etc.) which related to influenza virus entry and replication. Additional anti-virus pharmacological evaluation was performed to confirm the anti-influenza virus effects of the targeted-bonded small molecules. In order to accumulate the minor active compounds and to find the unexpressed natural products from the bioactive microorganisms, gene-mining and optimizing methods were applied including OSMAC and silent gene regulation. Particularly those containing synthetic terpene-PKS gene cluster and synthetic alkaloid hybrid gene cluster will be selected for activation to mine new structures with potential anti-influenza virus. In addition, the bioactive compounds with four different molecular scaffolds which had proved to possess anti-flu virus effects are selected for further investigation of their mechanistic pathways related to the targets of influenza virus entry, replication and release. The molecules with anti-drug resistant flu virus species and without resistant action with flu virus such as diketopiperazines and biphenyl esters will be examined in detail to find the new mechanism of drug resistant inhibition. Moreover, the potent antiviral molecules as the molecular probes will be selected for further investigation with proteomic and genomic methods to find new virus/host cell targets, which may insigne new antiviral pathway. Conclusively, following the research project, we intend to provide the marine microorganisms as a new source for the discovery of new antiviral agents and to overcome the resistant influenza virus which is unable to be treated by the drugs currently used for clinic trail.