A Platform to Identify Antifungal Compounds with Novel Action Mechanisms

项目摘要 社会需要新的化合物来防御真菌 病原体，其中许多病原体对现有疗法的抗性越来越强。抗真菌 化合物的发现已经被遗忘或忽视（参见2021年的综述出版物， 研究策略）。新的抗真菌化合物的最佳可能来源之一 具有潜在的新作用机制的是丝状真菌， 微生物生命的最大多样性这项研究计划推进了 宏基因组学，以证明构巢曲霉既是异源宿主， 初步筛选抗真菌靶标，结合RNA测序和真菌病原体 筛选真菌生物合成基因簇（BGC）和基因组，并发现 新的抗真菌化学品，并确定最佳的主要候选人的临床 发展Intact Genomics和威斯康星州麦迪逊大学的科学家们 结合了四项关键技术突破， 用于筛选小分子的范例。真菌人工合成的研究进展 染色体（FAC）工具包括：1）用于异源染色体的改进的方法学， 全长BGC-FAC的表达; 2）表达BGC-FAC的FAC异源菌株 抗真菌化合物也表现出异常表型; 3）新的作用机制 通过RNA深度测序发现的异常表型BGC-FACs; 4）a 一组真菌病原体的快速和改进的筛选方法，以确定新 抗真菌化合物。第一阶段SBIR将建立在以前成功的基础上， 通过筛选FAC中的抗真菌化合物进行研究。我们将描述 通过BGC-FAC克隆和FAC文库表达的抗真菌剂，以确定 临床开发的最佳主要候选人。主要候选人将有新的化学 结构，对多种真菌病原体具有高效力，并且毒性最小 针对人类红细胞。每一个不同的技术所必需的， 建议的研究已被证明是有效的单独;因此，综合 这些不同的方法有很高的成功概率，也代表了 在发现抗真菌药物方面取得了重大进展。

Project Summary There is societal need for new compounds in our arsenal of defenses against fungal pathogens, many of which are increasingly resistant to existing therapeutics. Antifungal compound discovery has been forgotten or neglected (see a review publication 2021 at Research Strategy). One of the best possible sources for new antifungal compounds with potentially novel mechanisms of action is within filamentous fungi, which have the greatest diversity of microbial life. This research proposal advances the science of metagenomics, to demonstrate Aspergillus nidulans as both a heterologous host and an initial antifungal screening target, to integrate with RNA sequencing and fungal pathogen screening of fungal biosynthetic gene clusters (BGCs) and genomes, and to discover novel antifungal chemicals and identify the best lead candidates for clinical development. Scientists at Intact Genomics, and University of Wisconsin at Madison have combined four key technological breakthroughs that result in an improved paradigm for screening small molecules. The improvements in fungal artificial chromosome (FAC) tools include: 1) an improved methodology for heterologous expression of full-length BGC-FACs; 2) the FAC heterologous strains expressing antifungal compounds also showing abnormal phenotypes; 3) new action mechanisms of abnormal phenotype BGC-FACs to be uncovered by RNA deep sequencing; 4) a panel of fungal pathogens for rapid and improved screening method to identify novel antifungal compounds. This Phase I SBIR will build upon the success of previous research by screening FACs for antifungal compounds. We will characterize the antifungal agents expressed by BGC-FAC clones and FAC libraries to determine the best lead candidates for clinical development. Lead candidates will have novel chemical structures, have high potency against multiple fungal pathogens, and minimal toxicity against human red blood cell. Each of the different technologies necessary for the proposed research has been proven effective separately; therefore, the synthesis of these different methods has a high probability of success and also represents a significant advancement for the science of antifungal discovery.