Protein Structure/Function by NMR, Crystallography and Computational Chemistry

通过 NMR、晶体学和计算化学研究蛋白质结构/功能

• 批准号: 7558297
• 负责人: Eric Oldfield
• 金额: $ 36.59万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7558297
• 关键词: Affect; Anabolism; Anti-Bacterial Agents; Antibiotics; Antifungal Agents; Area; Azoles; Bacteria; Binding; Bone Resorption; Carotenoids; Cell Wall; Cells; Cholesterol; Clinical; Clinical Data; Commit; Communicable Diseases; Communities; Community-Acquired Infections; Complex; Computer Simulation; Computing Methodologies; Confidential Information; Crystallography; Defense Mechanisms; Development; Dioxygenases; Diphosphates; Disease; Drug Delivery Systems; Drug Metabolic Detoxication; Drug resistance; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Escherichia coli; Funding; Gene Expression Profile; Geranyltranstransferase; Grant; Hospitals; Human; Immune; Immune system; Immunomodulators; Infection; Interferons; Isoprene; Language; Lead; Mediating; Metalloproteins; Methicillin Resistance; Miconazole; Molecular Conformation; Morphology; Mus; NBL1 gene; Natural Immunity; Oranges; Osteoporosis; Pharmaceutical Preparations; Physical condensation; Pigments; Process; Protozoa; Public Health; Quantitative Structure-Activity Relationship; Reactive Oxygen Species; Reporting; Research; Resistance; Roentgen Rays; Shigella flexneri; Squalene Synthetase; Staphylococcus aureus; Structure; Structure-Activity Relationship; System; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; TNF gene; Techniques; Testing; Trypanosoma brucei brucei; Trypanosoma cruzi; Virulence; Virulence Factors; Virulent; Vitamin K 2; Work; Zoledronate; base; bisphosphonate; cancer immunotherapy; cancer therapy; cell growth; cell killing; computational chemistry; computer studies; cytokine; decaprenyl pyrophosphate synthetase; dehydrosqualene; design; enzyme pathway; farnesyltranstransferase; guanidinium; hypercholesterolemia; immunoregulation; inhibitor/antagonist; interest; isopentenyl pyrophosphate; isoprenoid; killings; macrophage; neoplastic cell; neutrophil; novel; novel strategies; pre-clinical; protein structure function; public health relevance; resistant strain; response; small molecule; solid state nuclear magnetic resonance; staphyloxanthin; therapeutic target; undecaprenyl pyrophosphate synthetase

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.

描述（由申请人提供）：这项工作的广泛的总体目标是使用NMR、X射线和计算方法来帮助开发新型抗菌剂，除了直接杀死细菌外，还能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子类胡萝卜素毒力因子--葡萄球菌黄素形成的分子。在最近的工作中，我们发现人角鲨烯合酶抑制剂也可以阻断S.金黄色葡萄球菌，在200 nM水平。结果S。aureus（L.金黄色=金黄色）是白色的，在小鼠中是非感染性的，并且被嗜中性粒细胞杀死，因为它们降低了对活性氧的防御。在目标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S。金黄色葡萄球菌菌株对常规抗生素产生耐药性。第二个目标是开发目前用作抗真菌剂的唑类分子，作为抗S。金黄色葡萄球菌，阻断细菌黄素血红蛋白双加氧酶（从先天免疫细胞中解毒NO），以及影响类异戊二烯生物合成。我们将首先研究唑类化合物如何与黄素血红蛋白结合，并推导出结构-活性关系，以指导其他更有效的抑制剂的设计。其次，我们将研究唑类化合物如何发挥其直接的抗菌活性。我们建议测试的假设，这种活性是抑制类异戊二烯生物合成的结果，通过相关的类异戊二烯水平与抗菌活性，并通过使用微阵列技术来研究细菌的转录组。如果成功，这项工作将导致新的唑类化合物，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。第三个目标是开发双膦酸盐，激活？？除了开发直接杀死细菌的新型双膦酸盐外，先天免疫系统的T细胞还可以杀死细菌。在每一个目标中，我们将使用开发替代用途的策略，替代已经在人类中使用或测试的现有类型的药物：降低胆固醇的药物，阻断S。金黄色葡萄球菌（Aim 1）、具有抗菌活性的抗真菌唑类（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，用两三句话，用通俗的语言描述本研究与公共卫生的相关性。如果申请获得资助，此描述将成为公共信息。因此，不包括专有/机密信息。公共卫生相关性拟议的研究旨在导致新的方法来治疗传染病。将特别强调开发现有类型药物的替代，新用途：降低胆固醇的分子，也可以阻止葡萄球菌感染;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。