HTS assays for the methylerythritol-4-phosphate pathway

4-甲基赤藓糖醇-4-磷酸途径的 HTS 测定

基本信息

批准号：
8373656
负责人：
DEAN C CRICK
金额：
$ 39.92万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8373656
关键词：
Anabolism Antibiotics Antitubercular Agents Area Attention Bacteria Biochemical Biological Assay Carboxy-Lyases Cells Clinical Communities Development Diphosphates Drug Delivery Systems Drug Design Drug Resistant Tuberculosis Engineering Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Eukaryotic Cell Extreme drug resistant tuberculosis Genes Genome Genus Mycobacterium Goals Gram-Negative Bacteria Growth Human Incidence International Knowledge Laboratories Lead Mevalonate kinase Multidrug-Resistant Tuberculosis Mus Mycobacterium tuberculosis NIH Program Announcements Operon Orthologous Gene Pathway interactions Pharmaceutical Chemistry Pharmaceutical Preparations Phosphomevalonate kinase Physiological Plasmodium Property Proteins Reaction Role Running Screening procedure Series Solutions Source Testing Therapeutic Toxic effect Toxoplasma Tuberculosis Work Yeasts antimicrobial base drug development enzyme pathway fosmidomycin genetic evolution high throughput screening in vivo inhibitor/antagonist inorganic phosphate interest isopentenyl pyrophosphate isoprenoid killings mevalonate novel pathogen pathogenic bacteria programs response small molecule tool tuberculosis drugs tuberculosis treatment working group

项目摘要

DESCRIPTION (provided by applicant): This proposal is based on studies of the methylerythritol-4-phosphate (MEP) pathway that synthesizes the universal precursors of isoprenoid compounds, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), in which we have identified potential drug targets in Mycobacterium tuberculosis and other bacterial pathogens. These targets also exist in Plasmodium and Toxoplasma species. The enzymes involved in the MEP pathway in pathogenic bacteria have recently generated a great deal of attention as a source of targets for novel antimicrobials; all enzymes in the MEP pathway are thought to be potential targets for novel drugs as the MEP pathway has been demonstrated to be essential in both Gram-positive and Gram-negative bacteria and human cells do not contain orthologs. In addition, a natural antibiotic, fosmidomycin, which inhibits the second enzyme in the pathway (IspC) also inhibits the growth of Gram-negative bacteria and we have shown that inhibitors of the first enzyme in the MEP pathway (Dxs) also inhibits the growth of M. tuberculosis. However, few laboratories have been able to pursue this topic in detail due to a lack of substrates and appropriate HTS amenable assays. Our previous work in this area was aimed at characterizing MEP pathway enzymes from bacterial pathogens and developing efficient syntheses for the substrates; our present goal is to identify and develop inhibitors of previously characterized enzymes to provide potential leads for development of novel antimicrobial compounds. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to develop high throughput screens to identify inhibitors of the methylerythritol 4- phosphate biosynthetic bacteria in Mycobacterium tuberculosis, which will potentially generate new lead compounds and eventually new drugs for the treatment of tuberculosis.

描述（由申请人提供）：该提案基于对 4-磷酸甲基赤藓糖醇 (MEP) 途径的研究，该途径合成类异戊二烯化合物的通用前体、异戊烯基二磷酸 (IPP) 和二甲基烯丙基二磷酸 (DMAPP)，我们在其中鉴定了结核分枝杆菌和其他细菌病原体的潜在药物靶点。这些目标也存在于疟原虫和弓形虫物种中。病原菌 MEP 途径中涉及的酶作为新型抗菌药物的靶标来源，最近引起了广泛关注。 MEP 途径中的所有酶都被认为是新药的潜在靶标，因为 MEP 途径已被证明在革兰氏阳性和革兰氏阴性细菌中都是必需的，并且人类细胞不包含直系同源物。此外，抑制途径中第二种酶（IspC）的天然抗生素磷米霉素也能抑制革兰氏阴性菌的生长，我们已经证明MEP途径中第一种酶（Dxs）的抑制剂也能抑制革兰氏阴性菌的生长。结核分枝杆菌。然而，由于缺乏底物和合适的高温超导检测方法，很少有实验室能够详细研究这一主题。我们之前在该领域的工作旨在表征细菌病原体的 MEP 途径酶并开发底物的有效合成方法；我们目前的目标是识别和开发先前表征的酶的抑制剂，为新型抗菌化合物的开发提供潜在的线索。公共卫生相关性：该提案的目标是开发高通量筛选，以鉴定结核分枝杆菌中甲基赤藓糖醇 4-磷酸生物合成细菌的抑制剂，这将有可能产生新的先导化合物，并最终产生用于治疗结核病的新药物。