Fungal natural products targeting antimicrobial resistant Mycoplasma genitalium

性传播感染是一个全球性的健康问题，每天有超过100万人感染。 生殖支原体是一种重要的性传播细菌病原体，与严重的上尿路感染有关， 妇女生殖道后遗症和男女下生殖道感染。2019年11月， 疾病预防控制中心召开了一个研讨会，重点是性病相关的盆腔炎性疾病，其中M。生殖器是 被认为是一种关键的，但研究不足的新兴病原体，威胁着美国人的安全和福利， 公众M.在美国，生殖器感染率高于淋病奈瑟菌和沙眼衣原体 在越来越多的对推荐的一线治疗阿奇霉素耐药的情况下， 在许多情况下为50%，在高危人群中达到70 - 90%。令人担忧的是，对阿托沙星的耐药性， 推荐用于大环内酯类耐药菌株，越来越多地被检测到。这促使CDC将M. genitalium 在“2019年美国抗生素耐药性威胁”报告中的观察名单上。需要新的药物来 针对临床上正在演变成几乎无法治愈的病原体。我们假设真菌的天然产物 是一种尚未开发的新型抗菌剂来源，可用于靶向M。生殖器并提议将联合收割机 我们在药物发现和M.生殖器生物学（美国仅有的几个研究生殖器生物学的实验室之一） 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， 通过以下三个具体目标提供迫切需要的生物活性药物引线：（1） 真菌衍生的纯化合物和提取物用于抑制M.生殖器生长;（2）优先考虑生物活性 物质，进行抑制分子的测定指导纯化，并确定其结构;和（3） 研究生物活性物质对临床菌株的活性，以及耐药性发展试验。 具体地说，我们将使用900个纯化的真菌天然产物的文库，以及2,500个从真菌中提取的提取物。 在广泛的地理和环境地点收集的分类多样的真菌活动 分支杆菌生殖器（菌株G37将用作试验传感器菌株）。高活性纯化合物， 对人细胞表现出低水平毒性的细胞将进一步针对低传代细胞的集合进行测试。 抗微生物M.生殖器临床分离株。在表征活性成分的抑制曲线后， 化合物作为杀菌剂或抑菌剂，将探讨耐药性发展的潜力。我们 预计在R21研究的每年期间提供2 - 3种新的生物活性天然产物支架， 目标是鉴定高度有前途的分子，这些分子将成为进一步的先导化合物的主题， 在未来R01研究的背景下进行开发。我们不知道以前曾有系统地集中努力 真菌及其天然产物作为M.生殖器，我们对这个结果感到非常鼓舞 我们的初步研究表明，某些真菌代谢产物和提取物显示出有效的抑制作用， 对抗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.