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

基本信息

批准号：
7754041
负责人：
Eric Oldfield
金额：
$ 36.16万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
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/7754041
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 Computing Methodologies Confidential Information Crystallography Defense Mechanisms Development Dioxygenases Diphosphates Drug Metabolic Detoxication Drug resistance Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Funding Gene Expression Profile Grant Hospitals Human Immune Immune system Immunomodulators Infection Language Lead Mediating Metalloproteins Methicillin Resistance Miconazole Molecular Conformation Morphology Mus NBL1 gene Names Oranges Osteoporosis Pharmaceutical Preparations Physical condensation Pigments Principal Investigator Process Public Health Quantitative Structure-Activity Relationship Reactive Oxygen Species Research Resistance Roentgen Rays Squalene Synthetase Staphylococcus aureus Structure Structure-Activity Relationship System T-Cell Activation T-Cell Receptor T-Lymphocyte Techniques Testing Virulence Virulence Factors Virulent Vitamin K 2 Work Zoledronate base bisphosphonate cancer immunotherapy cell killing computational chemistry computer studies cytokine decaprenyl pyrophosphate synthetase dehydrosqualene design farnesyltranstransferase guanidinium hypercholesterolemia immunoregulation inhibitor/antagonist interest isopentenyl pyrophosphate isoprenoid killings macrophage neoplastic cell neutrophil novel novel strategies pre-clinical programs protein structure function public health relevance resistant strain response small molecule solid state nuclear magnetic resonance staphyloxanthin therapeutic target undecaprenyl pyrophosphate synthetase

项目摘要

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）：这项工作的广泛目标是使用NMR，X射线和计算方法来帮助开发新型的抗细菌剂，从而增强了先天免疫系统的杀戮，除了直接杀死细菌外。第一个目的是开发抑制金黄色葡萄球菌中橙色类胡萝卜素毒力因子（葡萄糖糖）形成的分子。在最近的工作中，我们发现人鳞状合酶抑制剂还可以在200 nm水平上阻止金黄色葡萄球菌的葡萄糖糖果生物合成。产生的金黄色葡萄球菌（L.金黄色葡萄球菌=黄金）是白色的，在小鼠中无感染，被嗜中性粒细胞杀死，因为它们减少了对活性氧的防御能力。在AIM 1中，我们将使用NMR，X射线和QSAR结果开发更有效的化合物来指导设计过程。如果成功的话，鉴于越来越多的S.金黄色菌株对常规抗生素具有抗性。第二个目的是开发当前用作抗神经链球菌的azole类别的分子，作为针对金黄色葡萄球菌的药物，阻断细菌flavohemoglobin二加氧酶（从先天性免疫细胞中排毒NO），以及影响类异磷酸生物合成。我们将首先研究Azoles如何结合氟珠蛋白并推断结构活性关系，以指导其他更有效的抑制剂的设计。其次，我们将研究硫唑如何发挥其直接的抗菌活性。我们建议通过将类吸收剂与抗细菌活性相关联，并使用微阵列技术研究细菌转录组来检验这种活性是抑制类异丙生素生物合成的结果的假设。如果成功的话，这项工作将导致新型的硫唑抑制细菌防御剂，以免基于基于杀害的杀戮，以及抑制细菌细胞壁生物合成的新化合物。第三个目的是开发激活的双膦酸盐？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细胞可杀死细菌。在每个目标中，我们都将使用为已经在人类中使用或测试过的现有类型的药物开发替代用途的策略：降低胆固醇的药物，这些药物可以阻止金黄色葡萄球菌（AIM 1），具有抗细菌活性的抗真菌硫唑则（AIM 2）（AIM 2）和具有免疫性或直接抗菌活性的骨骼（AIM）（AIM 3）或三分之二（AIM 3）。这项研究对公共卫生。如果申请是资助的，那么此描述将成为公共信息。因此，请勿包含专有/机密信息。公共卫生相关性该研究旨在导致治疗传染病的新方法。将特别强调为现有类型的药物开发替代的新颖用途：降低胆固醇的分子，这些分子也阻断了葡萄球菌感染；具有抗细菌活性的抗神经和刺激免疫系统并直接杀死细菌的骨吸收药物。