Optimization of the Antifungal Properties of the Lanthipeptide Pinensin and Search for Naturally Occurring Homologs

羊毛脂肽抗真菌特性的优化及天然同系物的搜索

• 批准号: 525824070
• 负责人: Dr. Julian Hegemann
• 金额: --
• 依托单位: Helmholtz-Institut für Pharmazeutische Forschung Saarland
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/525824070?language=en
• 关键词: Optimization; Antifungal; Properties; Lanthipeptide; Pinensin

Natural products are an important source of new anti-infectives and novel drug leads. Amongst these secondary metabolites, ribosomally synthesized and post-translationally modified peptides (RiPPs) represent an emerging superfamily that stirs up increasing interest due to their intriguing bioactivities, structural diversity, and remarkable sequence plasticity that facilitates their use for structure-activity-relationship studies and as bioengineering scaffolds. The development of new anti-infective agents is an increasingly urgent matter due the rise of antibiotic resistances and RiPPs often exhibit promising anti-infective activities, which warrants to take a closer look at them in the search for novel drug leads in the fight against infectious diseases. Such efforts are greatly facilitated by the fact that RiPP precursors are genetically encoded, which enables rapid compound diversification by simple mutation of the precursor-encoding gene in a suitable heterologous production system. The main aim of the proposed research project is the development of new antifungal drug leads starting from the natural product pinensin as lead molecule. The class I lanthipeptide pinensin is the first and so far only antifungal lanthipeptide reported in the literature. It exhibits a broad activity against pathogenic fungi, albeit with cytotoxic side effects that so far have prevented its use in a clinical setting. Due to the high demand for new antifungal drugs and previous studies demonstrating how other compounds with similar initial problems could be optimized successfully to obtain new antifungal agents, the planned project is timely and of high significance. The goal is to improve the therapeutic index of the lead molecule through structure-activity-relationship studies for paving the way for its potential use as a therapeutic. The structure-activity-relationship studies will be complemented by genome mining targeted at isolating closely related, naturally occurring homologs in search for superior lead structures. Preliminary genome mining data already suggests that certain pinensin homologs might play roles as virulence factors of clinically relevant pathogens with antibiotic resistances, which provides an additional layer that underlines the significance of the planned project. In preparation of this proposal, a system that enables the heterologous production of pinensin in E. coli has already been successfully established. Henceforth, the work on this project will begin with the further optimization of said production system for maximizing compounds yields. This in turn will facilitate the generation of an extensive library of pinensin variants aiming at the optimization of the selectivity of the antifungal versus the cytotoxic activities. In addition, mode-of-action studies are planned that will be utilizing the generation and sequencing of resistant mutants, pull-down assays, and fluorescence microscopy based localization studies.

天然产物是新型抗感染药物和新药先导的重要来源。在这些次级代谢产物中，核糖体合成和后修饰的肽（RIPPs）代表了一个新兴的超家族，由于其有趣的生物活性，结构多样性和显着的序列可塑性，促进其用于结构-活性-关系研究和生物工程支架，激起了越来越多的兴趣。由于抗生素耐药性的增加，新的抗感染药物的开发是一个越来越紧迫的问题，RIPP通常表现出有前途的抗感染活性，这值得在寻找新的药物领导人在对抗感染性疾病的斗争中仔细研究它们。RiPP前体被遗传编码的事实极大地促进了这种努力，这使得能够通过在合适的异源生产系统中简单突变编码RiPP的基因来实现快速的化合物多样化。该研究项目的主要目的是从天然产物松能素作为先导分子开始开发新的抗真菌药物先导物。I类羊毛硫肽松能菌素是第一个也是迄今为止唯一一个在文献中报道的抗真菌羊毛硫肽。它对致病真菌表现出广泛的活性，尽管具有细胞毒性副作用，迄今为止阻止了其在临床环境中的使用。由于对新的抗真菌药物的高需求，以及先前的研究表明如何成功优化具有类似初始问题的其他化合物以获得新的抗真菌药物，因此计划的项目是及时的，具有重要意义。目标是通过结构-活性-关系研究来提高先导分子的治疗指数，为其作为治疗剂的潜在用途铺平道路。结构-活性-关系研究将通过基因组挖掘得到补充，基因组挖掘旨在分离密切相关的天然同源物，以寻找上级先导结构。初步的基因组挖掘数据已经表明，某些pinensin同系物可能作为具有抗生素耐药性的临床相关病原体的毒力因子发挥作用，这提供了一个额外的层面，强调了计划项目的重要性。在准备这项建议时，一个系统，使异源生产松素在E。大肠杆菌已经成功建立。尽管如此，该项目的工作将开始，进一步优化所述生产系统，以使化合物产率最大化。这反过来又将促进广泛的松素变体库的生成，旨在优化抗真菌活性与细胞毒性活性的选择性。此外，计划进行作用模式研究，将利用耐药突变体的生成和测序、下拉试验和基于荧光显微镜的定位研究。