Reversing Antifungal Drug Resistance

描述(申请人提供)：由医学相关微生物获得的耐药性对人类健康构成严重威胁，并具有巨大的经济后果。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，并且在分子水平上分享许多相同的机制，这些机制支持组成人类宿主的细胞的生长和生存。在真菌中具有不同靶点的药物类别的数量非常有限，当前抗真菌药物的有效性受到损害，原因要么是对接受药物的患者有重大毒性，要么是经常出现高度耐药。该项目的目标是发现能够逆转真菌耐药性的新化合物，从而阐明致病真菌耐药性的机制，并使现有的抗真菌药物更安全和更有效。为了实现这一雄心勃勃的目标，将与美国国立卫生研究院(NIH)分子图书馆探针生产中心网络(MLPCN)内的一个指定中心合作，实施一项旨在实现以下具体目标的研究计划：目标1：优化并然后执行数十万种单独化学品的高通量机器人筛选，以寻找能够逆转从接受非常常用的抗真菌药物氟康唑的患者分离的真菌菌株的抗药性的化合物。目标2：通过测量在初级筛选中确定的化合物的效力、它们对各种真菌的活性光谱来评价它们，它们对人类和真菌细胞的选择性，并确定它们逆转抗真菌耐药性的一般方式目标3：选择最有希望的10种化合物，并为每种化合物合成一组衍生物，以优化其抗真菌效力和特异性。该项目将把我们在利用遗传和生化技术研究真菌分子生物学方面的长期专业知识与MLPCN中心的卓越资源及其在高通量筛选技术和药物化学方面的专业知识结合起来。与最近被指定为MLPCN中心的布罗德研究所合作，已经开发并验证了基本的初步筛查试验。因此，这个短暂的一年项目的可交付成果预计将是几种用于探索真菌生物学的高度有用的化合物。这些探针对于我们和其他人研究真菌耐药性的机制将是无价的。除了它们的基础研究应用，在未来的工作中，它们的治疗相关性可以很容易地使用已建立的动物模型来评估。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发挥作用，在解决我们社会面临的致病微生物获得耐药性这一日益严重的问题方面可能被证明是唯一有效的。 公共卫生相关性：致病微生物获得的抗药性对人类健康构成日益严重的威胁，并给我们的卫生保健系统带来巨大的经济负担。该项目将使用最先进的筛选技术来发现能够逆转人类致病真菌耐药性的化合物。由于它们将被发现的新方式，这些化合物中的许多可能以以前未知和未开发的方式发挥作用，在解决我们社会面临的获得性抗生素耐药性这一严重问题方面可能被证明是唯一有效的。

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.