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: 8212022
• 负责人: Eric Oldfield
• 金额: $ 35.67万
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8212022
• 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; Geranyltranstransferase; Grant; Health; Hospitals; Human; Immune; Immune system; Immunomodulators; Infection; Language; Lead; Mediating; Metalloproteins; Methicillin; 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; TNF gene; 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; 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）：这项工作的广泛总体目标是使用核磁共振、X 射线和计算方法来帮助开发新型抗菌剂，除了直接杀死细菌之外，还可以增强基于先天免疫系统的杀伤力。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素形成的分子。在最近的工作中，我们发现人类角鲨烯合酶抑制剂也可以以 200 nM 的水平阻断金黄色葡萄球菌中葡萄球菌黄素的生物合成。产生的金黄色葡萄球菌（金黄色葡萄球菌=金色）是白色的，对小鼠无感染性，并被中性粒细胞杀死，因为它们对活性氧的防御能力下降。在目标 1 中，我们将开发更有效的化合物，利用 NMR、X 射线和 QSAR 结果来指导设计过程。如果成功的话，鉴于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，这项工作将具有重要意义。第二个目标是开发目前用作抗真菌剂的唑类分子，作为抗金黄色葡萄球菌的药物，阻断细菌黄素血红蛋白双加氧酶（其解毒先天免疫细胞中的 NO），以及影响类异戊二烯生物合成。我们将首先研究唑类如何与黄素血红蛋白结合，并推断出结构-活性关系，这将指导其他更有效的抑制剂的设计。其次，我们将研究唑类如何发挥其直​​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，来检验这种活性是类异戊二烯生物合成抑制的结果的假设。如果成功，这项工作将产生抑制细菌防御一氧化氮杀伤作用的新型唑类化合物，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够激活 ?? 的双膦酸盐。先天免疫系统的 T 细胞可以杀死细菌，此外还可以开发直接杀死细菌的新型双磷酸盐。在每个目标中，我们将采用开发已在人体中使用或测试的现有类型药物的替代用途的策略：阻断金黄色葡萄球菌毒力的降胆固醇药物（目标 1）、具有抗菌活性的抗真菌唑类药物（目标 2）以及具有免疫调节或直接抗菌活性的骨吸收药物（目标 3）。此外，用两到三个句子，用简单、通俗的语言描述以下内容的相关性： 这项研究对公共卫生。如果申请获得资助，该描述将按原样成为公共信息。因此，请勿包含专有/机密信息。公共卫生相关性 拟议的研究旨在开发治疗传染病的新方法。将特别强调开发现有药物类型的替代、新用途：也可阻止葡萄球菌感染的降胆固醇分子；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。

项目成果

Insights into the mechanism of the antibiotic-synthesizing enzyme MoeO5 from crystal structures of different complexes.

• DOI: 10.1002/anie.201108002
• 发表时间: 2012-04-23
• 期刊: ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
• 影响因子: 16.6
• 作者: Ren, Feifei;Ko, Tzu-Ping;Feng, Xinxin;Huang, Chun-Hsiang;Chan, Hsiu-Chien;Hu, Yumei;Wang, Ke;Ma, Yanhe;Liang, Po-Huang;Wang, Andrew H. -J.;Oldfield, Eric;Guo, Rey-Ting
• 通讯作者: Guo, Rey-Ting

Head-to-head prenyl tranferases: anti-infective drug targets.

• DOI: 10.1021/jm300208p
• 发表时间: 2012-05-10
• 期刊: JOURNAL OF MEDICINAL CHEMISTRY
• 影响因子: 7.3
• 作者: Lin, Fu-Yang;Liu, Yi-Liang;Li, Kai;Cao, Rong;Zhu, Wei;Axelson, Jordan;Pang, Ran;Oldfield, Eric
• 通讯作者: Oldfield, Eric

HIV-1 Integrase Inhibitor-Inspired Antibacterials Targeting Isoprenoid Biosynthesis.

HIV-1积分酶抑制剂启发的抗菌靶向类异丙生素生物合成。

• DOI: 10.1021/ml300038t
• 发表时间: 2012-04-03
• 期刊: ACS MEDICINAL CHEMISTRY LETTERS
• 影响因子: 4.2
• 作者: Zhang, Yonghui;Lin, Fu-Yang;Li, Kai;Zhu, Wei;Liu, Yi-Liang;Cao, Rang;Pang, Ran;Lee, Eunhae;Axelson, Jordan;Hensler, Mary;Wang, Ke;Molohon, Katie J.;Wang, Yang;Mitchell, Douglas A.;Nizet, Victor;Odfied, Eric
• 通讯作者: Odfied, Eric

Dual dehydrosqualene/squalene synthase inhibitors: leads for innate immune system-based therapeutics.

• DOI: 10.1002/cmdc.201100589
• 发表时间: 2012-04
• 期刊: CHEMMEDCHEM
• 影响因子: 3.4
• 作者: Lin, Fu-Yang;Zhang, Yonghui;Hensler, Mary;Liu, Yi-Liang;Chow, Ohn A.;Zhu, Wei;Wang, Ke;Pang, Ran;Thienphrapa, Wdee;Nizet, Victor;Oldfield, Eric
• 通讯作者: Oldfield, Eric

Role of rsbU and staphyloxanthin in phagocytosis and intracellular growth of Staphylococcus aureus in human macrophages and endothelial cells.

rsbU 和葡萄球菌黄质在人巨噬细胞和内皮细胞中金黄色葡萄球菌的吞噬作用和细胞内生长中的作用。

• DOI: 10.1086/606012
• 发表时间: 2009
• 期刊: The Journal of infectious diseases
• 作者: Olivier,AurélieC;Lemaire,Sandrine;VanBambeke,Françoise;Tulkens,PaulM;Oldfield,Eric
• 通讯作者: Oldfield,Eric