Fungal natural products targeting antimicrobial resistant Mycoplasma genitalium

性传播感染是一个世界性的健康问题，每天有100多万人感染。 生殖支原体是一种重要的性传播细菌病原体，与严重的 女性生殖道后遗症和男性和女性下生殖道感染。2019年11月， 美国疾病控制与预防中心召开了一次研讨会，重点讨论与性病相关的盆腔炎，其中生殖道分支杆菌是 被认为是一种关键的、但未被研究的新病原体，它威胁着美国人的安全和福祉 公开的。生殖支原体在美国的患病率高于淋球菌和沙眼衣原体 在越来越多对推荐的一线治疗阿奇霉素产生抗药性的环境中，超过 在许多情况下为50%，在高危人群中达到70%-90%。令人担忧的是，对莫西沙星的耐药性， 推荐用于大环内酯类抗药性菌株，正在越来越多地被检测到。这促使疾控中心将生殖器分枝杆菌纳入其中。 在《2019年美国抗生素耐药性威胁》报告中被列入观察名单。需要新药来治疗 以临床上演变成几乎无法治愈的病原体为目标。我们假设真菌的天然产物 是一种未开发的新型抗菌剂来源，可用于靶向生殖道分枝杆菌，并提议结合 我们在药物发现和生殖支原体生物学方面的独特专业知识(美国仅有的几个研究 这种挑剔的有机体)来开发早期的先导化合物。我们将检验这一假设，并努力实现 通过以下三个具体目标提供急需的生物活性药物先导：(1)测试文库 抑制生殖支原体生长的真菌来源的纯化合物和提取物；(2)优先考虑生物活性 物质，进行实验指导的抑制分子的纯化，并确定其结构；以及(3) 调查生物活性物质对临床菌株的活性，以及耐药发展试验。 具体地说，我们将使用一个包含900种纯化真菌天然产物的库，以及从 在广泛的地理和环境活动地点收集的不同分类的真菌 对生殖支原体(G37菌株将作为检测敏感菌株)。高活性的纯化合物 对人类细胞表现出低水平的毒性将进一步针对低传代的集合进行测试 耐药生殖支原体临床分离株。在表征了活性物质的抑制曲线之后 作为杀菌剂或抑菌剂的化合物，其耐药性发展的潜力将被探讨。我们 预计在R21调查期间每年交付2-3个新的生物活性天然产品支架 目标是确定将成为进一步先导化合物研究对象的极有希望的分子 在未来R01研究背景下的发展。我们没有意识到先前有重点的有系统地开采地雷的努力 真菌及其天然产物对生殖支原体的抑制作用，我们对研究结果感到非常鼓舞 我们的初步研究表明，某些真菌代谢物和提取物具有很强的抑制作用 针对这种难以治疗的性传播寄生虫的活动。

Sexually transmitted infections are a worldwide health concern, with over a million infections acquired each day. Mycoplasma genitalium is an important sexually transmitted bacterial pathogen, associated with severe upper reproductive tract sequelae in women and lower genital tract infections in both men and women. In November 2019, the CDC convened a workshop focused on STD-related pelvic inflammatory diseases in which M. genitalium was recognized as a key, but understudied, emerging pathogen that threatens the safety and welfare of the American public. The prevalence of M. genitalium in the US is higher than Neisseria gonorrhoeae and Chlamydia trachomatis in a growing number of settings with resistance to the recommended first-line treatment, azithromycin, exceeding 50% in many situation and reaching 70-90% in the most-at-risk populations. Alarmingly, resistance to moxifloxacin, recommended for macrolide-resistant strains, is increasingly detected. This spurred the CDC to include M. genitalium on its watch list in the “2019 Antibiotic Resistance Threats in the United States” report. New drugs are needed to target what is clinically evolving into a nearly untreatable pathogen. We hypothesize that fungal natural products are an untapped source of novel antimicrobials that can be used to target M. genitalium and propose to combine our unique expertise in drug discovery and M. genitalium biology (one of only a few labs in the US that studies this fastidious organism) to develop early-stage lead compounds. We will test this hypothesis and work toward providing urgently needed bioactive drug leads through the following three specific aims: (1) test libraries of fungus-derived pure compounds and extracts for inhibition of M. genitalium growth; (2) prioritize bioactive substances, conduct assay-guided purification of inhibitory molecules, and determined their structures; and (3) investigate the activity of bioactive substances against clinical strains, as well as test for resistance development. Specifically, we will use a library of 900 purified fungal natural products, as well as 2,500 extracts produced from taxonomically diverse fungi collected over a wide range of geographical and environmental sites for activity against M. genitalium (strain G37 will be used as the assay sensor strain). Highly active pure compounds that exhibit low levels of toxicity toward human cells will be further tested against a collection of low passage antimicrobial-resistant M. genitalium clinical isolates. Upon characterizing the inhibition profiles of the active compounds as bactericidal or bacteriostatic, the potential for resistance development will be probed. We anticipate delivering 2-3 new bioactive natural-product scaffolds during each year of this R21 investigation with the goal of identifying highly promising molecules that will become the subjects of further lead compound development in the context of future R01 studies. We are unaware of prior focused efforts to systematically mine fungi and their natural products for inhibitors of M. genitalium and we have been highly encouraged by the results of our preliminary studies demonstrating that certain fungal metabolites and extracts exhibit potent inhibitory activities against this difficult-to-treat sexually-transmitted parasite.