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: 7142806
• 负责人: Eric Oldfield
• 金额: $ 32.96万
• 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/7142806
• 关键词: Leishmania major; Trypanosoma brucei rhodesiense; Trypanosoma cruzi; X ray crystallography; alkyltransferase; antibacterial agents; antiparasitic agents; antiprotozoal agents; chemical registry /resource; chemical structure function; deprenyl; drug design /synthesis /production; drug screening /evaluation; enzyme inhibitors; geranyl compound; laboratory mouse; leishmaniasis; microorganism disease chemotherapy; molecular cloning; nuclear magnetic resonance spectroscopy; phosphonate; pyrophosphates; thermodynamics; trypanosomiasis

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，整体目标是通过使用X射线晶体学，NMR光谱，量热法和细胞生长抑制分析，定量相关性（定量关系），定量，定量，定量，定量，定量，，使用X射线晶体光谱学，量热法和细菌的组合，靶向原生动物和细菌中异戊二烯生物合成途径的新型抗活性剂。和动物测试。第一个具体目的是使用X射线晶体学和固态NMR光谱法来研究新型抑制剂的结构，主要是Cruzi的Farnesyl Diphosphate合酶（FPPS），来自Cruzi的Farneyl diphosoma synthate（FPPS）金黄色葡萄球菌以及人类FPP（在设计抑制剂选择性的背景下感兴趣的双膦酸盐药物的靶标）。固态NMR将通过提供动力学（从2H NMR）和质子化状态信息（从13c，15n，15n，31p化学位移）来补充晶体学结果，以用于QSAR研究。第二个具体目的是研究结构功能关系。我们将通过使用等温滴定量热法和差异扫描来研究抑制剂结合，并使用经典的酶抑制技术来推导配体结合常数（KB）和KIS，以及使用抑制剂和抑制性的bonemottic bongormottion nym bongormits bongormits和bys bonemitign的抑制作用的系统来推导配体结合常数（KB），并使用抑制性的系统进行了抑制信息。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FPP抑制，以最大程度地减少骨吸附），以及使用非惯性QM描述符和2H NMR序列参数的使用，以增强这些方法的这些方法。第三个也是最终的特定目的是使用目标1和2的结果来设计，然后合成异戊二烯生物合成途径的特定抑制剂。这些抑制剂将包括新型的吡啶，磺硫酸和磷的抑制剂，旨在靶向在原生动物和细菌酶的活性位点发现的独特极性残基。然后，这些抑制剂将以生物利用形式进行配制，以用于动物测试。我们还将通过使用两种或三种化合物来关注协同或组合治疗方法，每种化合物靶向异戊二烯生物合成途径：已经观察到这种强烈的协同相互作用，现在需要使用新型抑制剂进行优化。 LIE摘要：提出的研究旨在导致新的治疗方法来治疗各种传染病。在美国，这些疾病主要是细菌性的，是主要的公共卫生威胁，而在世界其他地区，传染病主要由原生动物引起。这项工作旨在开发药物来治疗两种疾病。