Prenyldiphosphate Synthase Inhibitors: Novel Anti-Infective Agents

异戊二烯二磷酸合酶抑制剂：新型抗感染剂

• 批准号: 7686803
• 负责人: Eric Oldfield
• 金额: $ 31.87万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7686803
• 关键词: 1-deoxy-2-pentulose; Active Sites; Adsorption; African Trypanosomiasis; Anabolism; Animal Testing; Anti-Infective Agents; Area; Bacteria; Binding; Bioavailable; Biological Assay; Bone Resorption; Calorimetry; Cells; Chagas Disease; Chemicals; Combined Modality Therapy; Communicable Diseases; Complement; Crystallography; Cutaneous; Descriptor; Development; Differential Scanning Calorimetry; Disease; Drug Combinations; Drug Delivery Systems; Drug usage; Enzyme Inhibition; Enzymes; Escherichia coli; Explosion; Funding; Future; Geranyltranstransferase; Grant; Human; Information Systems; Investigation; Isoprene; Lead; Leishmania mexicana; Ligand Binding; Methods; Minerals; Modeling; Mus; NMR Spectroscopy; Parasites; Pathway interactions; Pharmaceutical Preparations; Protozoa; Public Health; Pyridium; Quantitative Structure-Activity Relationship; Reporting; Research; Resistance; Rest; Sorting - Cell Movement; Staphylococcus aureus; Structure; Structure-Activity Relationship; Techniques; Thermodynamics; Titrations; Trypanosoma brucei brucei; Trypanosoma cruzi; United States National Institutes of Health; Visceral Leishmaniasis; Work; bisphosphonate; bone; cell growth; combat; design; enzyme pathway; follow-up; high throughput screening; inhibitor/antagonist; inorganic phosphate; interest; neglect; novel; novel therapeutic intervention; protonation; quantum chemistry; solid state nuclear magnetic resonance; structural genomics; undecaprenyl pyrophosphate synthetase

DESCRIPTION (provided by applicant): The broad, overall objective of this work is to develop novel anti-infective agents which target the isoprene biosynthesis pathways in protozoa and bacteria by using a combination of x-ray crystallography, NMR spectroscopy, calorimetry, enzyme and cell growth inhibition assays, QSAR (quantitative structure-activity relationship) techniques, quantum chemistry, synthesis, drug delivery and animal testing. The First Specific Aim is to use x-ray crystallography and solid-state NMR spectroscopy to investigate the structures of novel inhibitors bound to, primarily, farnesyl diphosphate synthase (FPPS) from Trypanosoma cruzi (the causative agent of American trypanosomiasis or Chagas disease), Trypanosoma brucei (the causative agent of African sleeping sickness), E. coli and Staphylococcus aureus, as well as human FPPS (the target for the bisphosphonate drugs used in treating bone resorption diseases, of interest in the context of designing inhibitor selectivity). Solid-state NMR will be used to complement the crystallographic results by providing dynamics (from 2H NMR) and protonation state information (from 13C, 15N, 31P chemical shifts) for use in the QSAR investigations. The Second Specific Aim is to investigate structure-function relationships. We will investigate inhibitor binding by using isothermal titration calorimetry and differential scanning calorimetry, as well as by using classical enzyme inhibition techniques, to deduce ligand binding constants (KB) and Kis together with thermodynamics of binding information for the systems studied in Aim 1. In addition, inhibitor binding to human bone and bone mineral models will be investigated using chromatographic and NMR techniques. These results will all then be analyzed by using QSAR methods, including the use of differential QSAR methods (to optimize parasite/bacterial inhibition versus human FPPS inhibition, and to minimize bone adsorption), together with the use of non-conventional QM descriptors and 2H NMR order parameters, to enhance the predictive utility of these methods. The Third and Final Specific Aim is to use the results from Aims 1 and 2 to design and then synthesize specific inhibitors of the isoprene biosynthesis pathway. These inhibitors will include novel pyridium, sulfonium and phosphonium species and will be designed to target unique polar residues found in the active site of the protozoal and bacterial enzymes. These inhibitors will then be formulated in bioavailable forms for use in animal testing. We will also focus on synergistic or combination therapy approaches by using two or three compounds, each of which target the isoprene biosynthesis pathway: such strongly synergistic interactions have already been observed and now need to be optimized using the novel inhibitors. Lay Summary: The research proposed is designed to lead to new therapeutic approaches to treat a variety of infectious diseases. In the US, these diseases are primarily bacterial and are a major public health threat while in the rest of the world, infectious diseases are mainly caused by protozoa. This work seeks to develop drugs to treat both sorts of disease.

描述（由申请人提供）：这项工作的广泛总体目标是通过结合使用 X 射线晶体学、核磁共振波谱、量热法、酶和细胞生长抑制测定、QSAR（定量构效关系）技术、量子化学、合成、药物递送，开发针对原生动物和细菌中异戊二烯生物合成途径的新型抗感染剂 和动物测试。第一个具体目标是使用 X 射线晶体学和固态核磁共振波谱来研究主要与克氏锥虫（美洲锥虫病或恰加斯病的病原体）、布氏锥虫（非洲昏睡病的病原体）、大肠杆菌和法呢基二磷酸合酶（FPPS）结合的新型抑制剂的结构。 金黄色葡萄球菌以及人类 FPPS（用于治疗骨吸收疾病的双膦酸盐药物的靶标，在设计抑制剂选择性的背景下令人感兴趣）。固态 NMR 将通过提供动力学（来自 2H NMR）和质子化态信息（来自 13C、15N、31P 化学位移）来补充晶体学结果，以供 QSAR 研究使用。第二个具体目标是研究结构与功能的关系。我们将通过使用等温滴定量热法和差示扫描量热法以及使用经典的酶抑制技术来研究抑制剂的结合，以推导出配体结合常数 (KB) 和 Kis 以及目标 1 中研究的系统的结合信息的热力学。此外，将使用色谱和 NMR 研究抑制剂与人骨和骨矿物质模型的结合 技术。然后，这些结果将通过使用 QSAR 方法进行分析，包括使用差异 QSAR 方法（以优化寄生虫/细菌抑制与人类 FPPS 抑制相比，并最大限度地减少骨吸附），以及使用非常规 QM 描述符和 2 H NMR 有序参数，以增强这些方法的预测效用。第三个也是最后一个具体目标是利用目标 1 和 2 的结果来设计并合成异戊二烯生物合成途径的特异性抑制剂。这些抑制剂将包括新型吡啶、锍和鏻物种，旨在针对原生动物和细菌酶活性位点中发现的独特极性残基。然后，这些抑制剂将被配制为生物可利用的形式，用于动物测试。我们还将重点关注使用两种或三种化合物的协同或联合治疗方法，每种化合物都针对异戊二烯生物合成途径：这种强烈的协同相互作用已经被观察到，现在需要使用新型抑制剂进行优化。概要：所提出的研究旨在开发治疗各种传染病的新治疗方法。在美国，这些疾病主要是细菌性疾病，是主要的公共卫生威胁，而在世界其他地区，传染病主要由原虫引起。这项工作旨在开发治疗这两种疾病的药物。