基于质谱-分子网络策略的柳珊瑚来源真菌环肽活性先导物发现

Marine microorganisms with high biodiversity have proven to be a rich source for searching for secondary metabolites with novel structures and potential bioactivities. In recent years, marine microorganisms have been considered as a hot resource for the discovery of bioactive marine natural products and lead compounds. Unfortunately, a critical problem not overcomed by many methodologies has emerged accompanying with the research on the secondary metabolites from marine microorganisms. The known compounds have been isolated repeatedly, and the new molecules especially with trace amounts have been lost frequently. To face this challenge, we propose to find bioactive lead compounds of cyclodepsipeptides from gorgonian-derived fungi by MS/MS molecular networking strategy. Firstly, the mass data and mass clearage modes of the secondary metabolites should be collected by LC-MS/MS analysis of the cultivated products of the fungal strains. A viewable MS/MS molecular network should be established based on the mass characteristics and clearage patterns of the same type of compounds. Consequently, under the guidance of the established MS/MS molecular network, the new cyclodepsipeptides with angiogenesis activity should be isolated from the fermentation broth of gorgonian-derived fungi. By using the MS/MS molecular network, the trace new compounds would be predicted and discovered, and the bioactive lead compounds whould be obtained accurately with high efficiency. The results from this research would be valuable to the dereplication of the secondary metabolites and the discovery for lead compounds with novel structures from marine microorganisms.

海洋微生物具有丰富的物种多样性，能够产生结构新颖、活性显著的次级代谢产物，已成为活性海洋天然产物和药物先导化合物发现的热点资源。然而，随着对海洋微生物次级代谢产物研究的不断深入，重复分离获得已知化合物、丢失微量存在的新化合物等问题日益突出。本项目瞄准海洋天然产物领域面临的这一关键问题，以柳珊瑚来源真菌为研究对象，开展基于质谱-分子网络策略的环肽活性先导物发现研究。首先利用LC-MS/MS系统分析发酵产物中化合物质谱及其裂解模式，依据同类化合物的质谱特征及其裂解规律，构建可视化质谱-分子网络数据库（MS/MS Molecular Network）；进而在质谱-分子网络策略指导下，以具有促血管新生等活性的新颖环肽类化合物为目标，从发酵产物中预测、排重、追踪、发掘微量新颖化合物，精准、高效发现活性先导物。本项目研究结果对海洋微生物代谢产物的排重和新颖结构药物先导化合物的发现具有启示和借鉴价值。

海洋微生物具有丰富的物种多样性，能够产生结构新颖、活性显著的次级代谢产物，已成为活性海洋天然产物和药物先导化合物发现的热点资源。然而，随着对海洋微生物次级代谢产物研究的不断深入，重复分离获得已知化合物、丢失微量存在的新化合物等问题日益突出。..本项目瞄准海洋天然产物领域面临的这一关键问题，以柳珊瑚来源真菌为研究对象，开展了基于质谱-分子网络策略(MS/MS Molecular Network)的环肽活性先导物发现研究。利用LC-MS/MS系统分析真菌Penicillium chrysogenum，Aspergillus versicolor，Aspergillus sclerotiorum, Scopulariopsis brevicaulis等发酵产物中化合物质谱及其裂解模式，依据同类化合物的质谱特征及其裂解规律，构建可视化质谱-分子网络数据库；进而以新颖环肽类化合物为目标，在质谱-分子网络策略指导下，结合OSMAC策略、化学表观遗传修饰和基因组采掘技术，通过生物活性追踪，从发酵产物中预测、排重、追踪、发掘微量新颖化合物。本研究获得了多个系列环肽类新颖结构化合物，分离鉴定新环肽化合物17个，半合成获得系列新衍生物31个；通过生物活性筛选评价，发现了促血管生成(3个)、抗结核杆菌(7个)、抗肿瘤(1个)等活性化合物11个；分析了高活性化合物的构效关系，评价了体内外活性，初步探讨了作用机制，为药物先导化合物的发现提供了化合物基础。..本研究证明了质谱-分子网络(MS/MS Molecular Networking)技术是发掘海洋微生物新颖结构活性次级代谢产物的有效途径，对海洋微生物代谢产物的排重和新颖结构药物先导化合物的发现具有启示和借鉴价值。