Developing Metallo-Beta-Lactamase Inhibitors

开发金属-β-内酰胺酶抑制剂

基本信息

批准号：
9023565
负责人：
ROBERT A. BONOMO
金额：
$ 52.86万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9023565
关键词：
Active Sites Address Aminoglycosides Animal Model Antibiotic Resistance Antibiotics Bacteria Bacterial Infections Binding Binding Sites Biological Assay Carbapenems Centers for Disease Control and Prevention (U.S.)Chelating Agents Chemicals Clinical Clinical Treatment Communities Complex Development Diversity Library Drug usage Enterobacteriaceae Enzymatic Biochemistry Enzymes Escherichia coli Evolution Generations Genes Goals Health Hospitals Human Hybrids In Vitro Infection Investigation Ions Kinetics Klebsiella pneumonia bacterium Knowledge Lactamase Lactams Lead Libraries Ligands Link Macrolide-resistance Metals Methods Microbiology Mission Molecular Conformation Monobactams Mononuclear Multi-Drug Resistance Nightmare Organic Synthesis Patients Penicillins Pharmaceutical Chemistry Pharmaceutical Preparations Plasmids Process Protein Isoforms Proteins Public Health Reporting Resistance Rifampin Series Site Spectrum Analysis Structure Sulfamethoxazole Superbug Techniques Testing Therapeutic Time United States National Institutes of Health Work Writing X-Ray Crystallography Zinc bacterial resistance base beta-Lactamase beta-Lactams carbapenem-resistant Enterobacteriaceae clinically relevant colistin resistance combat design drug discovery drug resistant bacteria experience flexibility high throughput screening improved inhibitor/antagonist innovation metalloenzyme microbial microorganism multidisciplinary new therapeutic target novel novel therapeutics pre-clinical programs public health relevance screening stoichiometry structural biology targeted treatment tigecycline tool

项目摘要

DESCRIPTION (provided by applicant): ß-Lactams are drugs of vital importance for treatment of bacterial infections. However, the emergence of bacterial metallo-ß-lactamase (MBL) enzymes has enabled resistance to almost all clinically used drugs in this class (excepting the monobactams). Arguably, the most worrisome MBL to emerge is New Dehli Metallo-ß- lactamase-1 (NDM-1), which can hydrolyze all generations of bicyclic ß-lactams that have been tested, including penems, cephems, and carbapenems. NDM-1 has been identified in a number of different Enterobacteriaceae, and is community-acquired (not just nosocomial). The gene is encoded in plasmids capable of horizontal transfer that also confer resistance to macrolides, aminoglycosides, rifampicin, sulfamethoxazole, and monobactams. Therefore MBLs, and NDM-1 in particular, represent a significant emerging worldwide health threat. Despite its importance, drugs that target NDM-1, or any other MBL, are not known. At the time of writing, there are only ~11, relatively weak inhibitors reported for NDM-1, and inhibitors of the other most clinically-relevant MBLs are limited in efficacy and chemical diversity. Inhibitor development has been difficult, in part, due to the flexible active sites of these enzymes and the challenges associated with targeting metalloenzymes. This proposal will directly address this deficit by using innovative fragment-based drug discovery complimented with high-throughput screening approaches, detailed characterization of inhibitor binding, bioinorganic spectroscopy, kinetics, X-ray crystallography, and assays of microbial efficacy. We will conduct a deep investigation and optimization of two conserved features of MBL inhibitors: metal-binding groups and substituents that interact with the flexible loops of the protein. Aim 1 uses privileged chelator- fragment libraries to identify moieties effective for inhibition of NDM-1 and other clinically-relevant B1 lactamases. Aim 2 uses diversity screening to identify and optimize structurally divergent inhibitors to elucidate how the flexible NDM-1 binding sites accommodate different ligands. Aim 3 uses a battery of structural and spectroscopic techniques to characterize, re-design, and ultimately optimize these inhibitors and their interactions with NDM-1. An experienced team brings proficiency in organic synthesis, inhibitor screening, medicinal chemistry, enzymology, spectroscopy, structural biology, and microbiology to advance hits into potent NDM-1 inhibitors with known mechanisms of action and microbial efficacy. This work will make critical contributions to validating novel, potent, structurally-diverse, and biologically-useful NDM-1 inhibitors. The proposed work is innovative due to the application of a unique chelator-fragment discovery approach, the extensive inhibitor-target characterization relevant to many B1 lactamases, and the multidisciplinary team committed to developing much needed first-in-class inhibitors as tools and lead compounds for therapeutic design to counter the global threat of NDM-1.

描述（由申请人提供）：β-内酰胺是治疗细菌感染至关重要的药物，然而，细菌金属-β-内酰胺酶（MBL）的出现使得几乎所有临床使用的此类药物产生耐药性（除此之外）。可以说，最令人担忧的 MBL 是 New Dehli Metallo-ß-内酰胺酶-1 (NDM-1)，它可以水解已测试的所有世代的双环β-内酰胺，包括青霉烯类、头孢烯类和碳青霉烯类，已在许多不同的肠杆菌科细菌中鉴定出该基因，并且是社区获得性的（不仅仅是医院内的）。能够水平转移的质粒也赋予对大环内酯类、氨基糖苷类、利福平、磺胺甲恶唑和因此，MBL，尤其是 NDM-1，尽管具有重要意义，但在撰写本文时，针对 NDM-1 或任何其他 MBL 的药物尚不清楚。如图 11 所示，报道的 NDM-1 抑制剂相对较弱，而其他与临床最相关的 MBL 抑制剂的功效和化学多样性有限。抑制剂的开发一直很困难，部分原因是这些酶的活性位点灵活。以及相关的挑战该提案将通过使用创新技术直接解决这一缺陷。基于片段的药物发现与高通量筛选方法、抑制剂结合的详细表征、生物无机光谱学、动力学、X射线晶体学和微生物功效分析相结合，我们将对 MBL 抑制剂的两个保守特征进行深入研究和优化。：与蛋白质柔性环相互作用的金属结合基团和取代基 Aim 1 使用特权螯合剂片段库来鉴定可有效抑制 NDM-1 和其他物质的部分。目标 2 使用多样性筛选来识别和优化结构上不同的抑制剂，以阐明灵活的 NDM-1 结合位点如何适应不同的配体。目标 3 使用一系列结构和光谱技术来表征、重新设计和分析。最终优化这些抑制剂及其与 NDM-1 的相互作用。经验丰富的团队精通有机合成、抑制剂筛选、药物化学、酶学、光谱学、结构生物学、这项工作将为验证新型、有效、结构多样且具有生物学用途的 NDM-1 抑制剂做出重要贡献。由于应用了独特的螯合剂片段发现方法、与许多 B1 内酰胺酶相关的广泛抑制剂靶标表征，以及致力于开发急需的一流抑制剂作为工具和用于应对 NDM-1 全球威胁的治疗设计的先导化合物。