Antifungal discovery from previously uncultivated bacteria

摘要 金黄色念珠菌是一种耐多药酵母，可导致侵袭性感染和死亡。它很容易传播 住院病人和疗养院居民之间的关系。C.Auris对 免疫抑制患者和新冠肺炎患者。特别是关注一些C. 金黄色葡萄球菌对所有三种主要的抗真菌药物都有耐药性。因此，这种致命的病原体一直是 被疾病控制中心(CDC)列为紧急威胁，新的抗真菌药物 迫切需要。 大多数抗真菌药物都是从环境微生物产生的天然化合物中提取的。 然而，从这个来源筛选抗真菌药物已经不受欢迎，因为容易培养的 微生物被破坏了。然而，99%的环境微生物从未被 筛选是因为它们在正常的实验室条件下不会生长。为了解决这个问题，我们使用了我们的 IChip培养技术，将大量以前未培养的微生物集合在一起。IChip 使微生物能够在其自然环境(如土壤)中生长，使它们能够进行必要的生长 各种因素。因此，我们筛选了其他药物发现计划无法接触到的微生物。我们有 利用iChip技术发现了几种新的抗菌化合物，如teixobactin，它是 目前正在进行使IND能够治疗耐药细菌感染的研究。在这个项目中，我们将筛选我们的 独特的微生物集合，用于杀死C.Auris的化合物。 在初步研究中，我们筛选了一小部分iChip分离株(3000株)，以对抗金黄色葡萄球菌。由此 有限的筛选，我们已经发现了五种潜在的新型抗真菌药物，我们将在这个项目中进行研究。 此外，我们将筛选更多的分离株(30,000株)，以获得更有希望的 化合物。粗提物将首先从发酵中产生，并对金黄色葡萄球菌进行筛选。这个 产生抗真菌活性的化合物将从提取物中分离出来，以确定其化学成分。 新颖性，对其他重要真菌(包括抗药性菌株)的效力，细胞毒性，以及 抵抗。有希望的化合物将在真菌感染(播散性)的小鼠疗效模型中进行测试 念珠菌病)。表现出良好动物功效的化合物将被视为先导化合物及其全部 化学结构将被阐明。我们在这个项目结束时的目标是发现1-2种新的先导化合物 用于临床前开发，以对抗金黄色葡萄球菌和潜在的其他真菌感染。

ABSTRACT Candida auris is a multidrug-resistant yeast that can cause invasive infection and death. It spreads easily between hospitalized patients and nursing home residents. C. auris is especially hazardous to immunosuppressed patients and those hospitalized with Covid‐19. Particularly concerning, some strains of C. auris are resistant to all three major classes of antifungal drugs. As a result, this deadly pathogen has been classified as an Urgent Threat by the Centers for Disease Control (CDC), and new antifungal drugs are desperately needed. Most antifungal drugs are derived from natural compounds produced by environmental microorganisms. However, screening for antifungals from this source has fallen into disfavor, as the readily culturable microorganisms have been overmined. However, 99% of environmental microorganisms have never been screened because they will not grow under normal laboratory conditions. To address this problem, we used our iChip culturing technology to assemble a large collection of previously uncultured microorganisms. The iChip enables microorganisms to grow in their natural environment (e.g., soils), giving them access to essential growth factors. As a result, we screen microorganisms that are inaccessible to other drug discovery programs. We have used the iChip technology to discover several new antibacterial compounds such as teixobactin, which is currently in IND-enabling studies to treat drug-resistant bacterial infections. In this project we will screen our unique microbe collection for compounds that kill C. auris. In preliminary studies, we screened a small number of our iChip isolates (3,000) against C. auris. From this limited screening, we have already found five potentially novel antifungals, which we will pursue in this project. In addition, we will screen a much larger number of our isolates (30,000) to obtain additional promising compounds. Crude extracts will first be produced from fermentations and screened against C. auris. The compounds producing the antifungal activity will then be isolated from the extracts to determine their chemical novelty, potency against other important fungi (including drug-resistant strains), cytotoxicity, and frequency of resistance. Promising compounds will then be tested in a mouse efficacy model of fungal infection (disseminated candidiasis). Compounds that show good animal efficacy will be considered lead compounds and their full chemical structure will be elucidated. Our goal at the end of this project is to discover 1-2 novel lead compounds for preclinical development to combat C. auris and potentially other fungal infections.