Protein Structure/Function by NMR, Crystallography and Computational Chemistry

描述（由申请人提供）：这项工作的广泛，总体目标是使用NMR， x射线和计算方法来帮助开发新的抗菌药物，除了直接杀死细菌外，还可以增强基于先天免疫系统的杀伤。第一个目标是开发抑制金黄色葡萄球菌中橙色类胡萝卜素毒力因子葡萄黄质形成的分子。在最近的工作中，我们发现人角鲨烯合成酶抑制剂也可以阻断金黄色葡萄球菌200 nM水平的葡萄黄质生物合成。由此产生的金黄色葡萄球菌（金黄色葡萄球菌）是白色的，在小鼠中没有传染性，被中性粒细胞杀死，因为它们对活性氧的防御能力降低了。在Aim 1中，我们将开发更有效的化合物，使用NMR， x射线和QSAR结果来指导设计过程。如果成功，这项工作将是重要的，因为越来越多的金黄色葡萄球菌菌株对传统抗生素产生耐药性。第二个目标是开发目前用作抗真菌的唑类分子，作为抗金黄色葡萄球菌的药物，阻断细菌黄血红蛋白双加氧酶（从先天免疫细胞中解毒NO），以及影响类异戊二烯的生物合成。我们将首先研究唑类药物如何与黄血红蛋白结合，并推断出结构-活性关系，这将指导设计其他更有效的抑制剂。其次，我们将研究唑类药物如何发挥其直接抗菌活性。我们建议通过将类异戊二烯水平与抗菌活性相关联，并使用微阵列技术研究细菌转录组，来验证这种活性是抑制类异戊二烯生物合成的结果这一假设。如果成功，这项工作将导致新的氮唑抑制细菌对no基杀伤的防御，以及抑制细菌细胞壁生物合成的新化合物。第三个目标是开发能激活？？除了产生直接杀死细菌的新型双磷酸盐外，先天免疫系统的T细胞也可以杀死细菌。在每个Aim中，我们将使用开发替代人类已经使用或测试的现有类型药物的策略：阻断金黄色葡萄球菌毒力的降胆固醇药物（Aim 1），具有抗菌活性的抗真菌唑类药物（Aim 2）和具有免疫调节或直接抗菌活性的骨吸收药物（Aim 3）。此外，用两到三句话，用通俗易懂的语言描述这项研究与公共卫生的相关性。如果该应用程序获得资助，则此描述将成为公共信息。因此，不要包含专有/机密信息。与公共卫生的关系拟议的研究旨在找到治疗传染病的新方法。将特别强调开发现有药物的替代新用途：降低胆固醇的分子也可以阻断葡萄球菌感染；具有抗菌活性的抗真菌药物，以及刺激免疫系统并直接杀死细菌的骨吸收药物。

DESCRIPTION (provided by applicant): The broad, overall objectives of this work are to use NMR, X-ray and computational methods to help develop novel anti-bacterial agents, which enhance innate immune system based killing, in addition to killing bacteria directly. The First Aim is to develop molecules that inhibit formation of the orange carotenoid virulence factor, staphyloxanthin, in Staphylococcus aureus. In recent work, we discovered that human squalene synthase inhibitors can also block staphyloxanthin biosynthesis in S. aureus, at 200 nM levels. The resulting S. aureus (L. aureus=golden) are white, non-infective in mice and are killed by neutrophils, since they have decreased defenses to reactive oxygen species. In Aim 1, we will develop more effective compounds, using NMR, X-ray and QSAR results to guide the design process. If successful, this work would be of importance given the increasing number S. aureus strains that are becoming resistant to conventional antibiotics. The Second Aim is to develop the azole class of molecules currently used as anti-fungals, as agents against S. aureus, blocking bacterial flavohemoglobin dioxygenase (which detoxifies NO from innate immune cells), as well as affecting isoprenoid biosynthesis. We will first investigate how azoles bind to flavohemoglobin and deduce structure-activity relationships that will guide the design of other, more potent inhibitors. Second, we will investigate how azoles exert their direct anti-bacterial activity. We propose to test the hypothesis that this activity is a result of the inhibition of isoprenoid biosynthesis, by correlating isoprenoid levels with anti-bacterial activity, and by using microarray techniques to investigate the bacterial transcriptome. If successful, this work would lead to novel azoles that inhibit bacterial defenses against NO-based killing, as well as new compounds that inhibit bacterial cell wall biosynthesis. The Third Aim is to develop bisphosphonates that activate ?? T cells of the innate immune system to kill bacteria, in addition to developing novel bisphosphonates that kill bacteria directly. In each Aim, we will use the strategy of developing alternate uses for existing types of drugs already used or tested in humans: cholesterol lowering drugs that block virulence in S. aureus (Aim 1), anti-fungal azoles with anti- bacterial activity (Aim 2) and bone resorption drugs that have immunomodulation or direct anti- bacterial activity (Aim 3).In addition, in two or three sentences, describe in plain, lay language the relevance of this research to public health. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. PUBLIC HEALTH RELEVANCE The research proposed is designed to lead to new approaches to treating infectious diseases. Particular emphasis will be given to developing alternate, novel uses for existing types of drugs: cholesterol lowering molecules that also block Staph infections; anti-fungals with anti-bacterial activity, and bone resorption drugs that stimulate the immune system as well as kill bacteria directly.