HTS assays for the methylerythritol-4-phosphate pathway

这一提议是基于对合成多聚体的甲基-4-磷酸（MEP）途径的研究。 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），其中我们已经确定了结核分枝杆菌和其他细菌中的潜在药物靶点 病原体这些靶标也存在于疟原虫和弓形虫物种中。参与蛋白质合成的 MEP途径作为病原菌的一个来源靶点，近年来引起了人们的极大关注 MEP途径中的所有酶都被认为是新型药物的潜在靶点， MEP途径已被证明在革兰氏阳性菌和革兰氏阴性菌中都是必需的 而人类细胞不含直系同源物。此外，一种天然抗生素，磷霉素，它抑制了 途径中的第二种酶（IspC）也抑制革兰氏阴性菌的生长，我们已经证明 MEP途径中的第一种酶（Dxs）的抑制剂也抑制M的生长。结核 然而，由于缺乏底物， 适当的HTS适用测定。我们以前在这一领域的工作旨在表征MEP途径 酶从细菌病原体和开发有效的合成底物;我们目前的目标是 鉴定和开发先前表征酶的抑制剂，为开发提供潜在线索 新的抗菌化合物。

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.