Reversing Antifungal Drug Resistance

描述（由申请人提供）：医学相关微生物获得性耐药对人类健康构成严重威胁，并产生巨大的经济后果。真菌病原体构成了一个特别的挑战，因为它们与人类的关系比细菌更密切，并且在分子水平上具有许多相同的机制，这些机制支持构成其人类宿主的细胞的生长和存活。在真菌中具有不同靶点的药物类别的数量非常有限，并且由于对接受它们的患者具有显着毒性或频繁出现高级别耐药性，当前抗真菌药物的有效性受到损害。该项目的目标是发现能够逆转真菌耐药的新化合物，从而阐明致病真菌耐药的机制，并使现有的抗真菌药物更安全、更有效。为了实现这一雄心勃勃的目标，一项旨在实现以下具体目标的研究计划将与美国国立卫生研究院（NIH）分子文库探针生产中心网络（MLPCN）内的指定中心合作进行：优化并执行数十万种单独化学物质的高通量机器人筛选，以找到可以逆转从接受非常常用的抗真菌药物氟康唑的患者中分离出的真菌菌株耐药性的化合物。通过测量它们的效力，它们对各种类型真菌的活性谱，它们对人类和真菌细胞的选择性来评估在初级筛选中鉴定的化合物，并确定它们逆转抗真菌药物耐药性的一般方式。目的3：选择10个最有希望的化合物，并为每个化合物合成一组衍生物，以优化它们的抗真菌效力和特异性。该项目将结合我们长期以来利用遗传和生化技术研究真菌分子生物学的专业知识，以及MLPCN中心的优秀资源及其在高通量筛选技术和药物化学方面的专长。与最近被指定为MLPCN中心的Broad研究所合作，已经开发并验证了基本的初级筛选试验。因此，这个为期一年的项目有望产生几种非常有用的化合物，用于探索真菌生物学。这些探针将为我们和其他人研究真菌耐药机制提供宝贵的价值。除了它们的基础研究应用之外，它们的治疗相关性可以在未来的工作中很容易地使用已建立的动物模型进行评估。由于它们将被发现的方式，这些化合物中的许多可能以以前未知和未开发的方式起作用，这可能被证明是解决我们社会面临的日益严重的由致病微生物获得性耐药性问题的独特有效方法。

DESCRIPTION (provided by applicant): Acquired drug resistance by medically relevant microorganisms poses a grave threat to human health and has enormous economic consequences. Fungal pathogens pose a particular challenge because they are more closely related to humans than bacteria and share many of the same mechanisms at the molecular level that support the growth and survival of the cells comprising their human hosts. The number of drug classes that have distinct targets in fungi is very limited and the usefulness of current antifungal drugs is compromised owing to either significant toxicity for the patient receiving them or the frequent emergence of high grade resistance. The objective of this project is to discover new chemical compounds capable of reversing fungal drug resistance, thereby illuminating the mechanisms responsible for drug resistance in disease-causing fungi and making currently available antifungal safer and more effective. To achieve this ambitious goal, a research plan designed to achieve the following specific aims will be pursued in collaboration with a designated center within the National Institutes of Health (NIH) Molecular Libraries Probe Production Center Network (MLPCN): Aim 1: Optimize and then execute a high throughput robotic screen of hundreds of thousands of individual chemicals to find compounds that can reverse the resistance of a fungal strain that was isolated from a patient receiving the very commonly used antifungal drug fluconazole Aim 2: Evaluate the compounds identified in the primary screen by measuring their potency, their spectrum of activity against various types of fungus, their selectivity for human versus fungal cells and determine the general way in which they reverse antifungal drug resistance Aim 3: Select the 10 most promising compounds and synthesize a panel of derivatives for each one to optimize their antifungal potency and specificity. This project will combine our long established expertise in studying the molecular biology of fungi using genetic and biochemical techniques with the outstanding resources of an MLPCN center and its expertise in high throughput screening technology and medicinal chemistry. In collaboration with the Broad Institute which was recently designated an MLPCN center, the essential primary screening assay has already been developed and validated. As a result, the deliverable outcome from this brief one year project is expected to be several highly useful chemical compounds with which to probe fungal biology. These probes will be invaluable to us and others for studying the mechanisms underlying fungal drug resistance. In addition to their basic research applications, their therapeutic relevance can be readily evaluated using established animal models in future work. By virtue of the way in which they will be discovered, many of these compounds are likely to act in previously unknown and unexploited ways that could prove uniquely effective in addressing the ever increasing problem of acquired drug resistance by disease-causing microorganisms that confronts our society. PUBLIC HEALTH RELEVANCE: Acquired drug resistance by disease-causing microorganisms poses an escalating threat to human health and imposes an enormous economic burden on our health care system. This project will use state of the art screening technologies to discover chemical compounds that can reverse drug-resistance in human disease- causing fungi. By virtue of the novel way in which they will be discovered, many of these compounds are likely to act in previously unknown and unexploited ways that could prove uniquely effective in tackling the serious problem of acquired antibiotic resistance that confronts our society.