HTS assays for the methylerythritol-4-phosphate pathway

这一建议是基于对甲基赤藓糖醇-4-磷酸(MEP)途径的研究，该途径合成 异戊二烯基二磷酸(IPP)和二甲基烯丙基二磷酸酯的通用前体 (DMAPP)，其中我们已经确定了结核分枝杆菌和其他细菌的潜在药物靶点 病原体。这些靶标也存在于疟原虫和弓形虫物种中。参与这一过程的酶 最近，病原菌中的MEP通路作为靶点引起了极大的关注 对于新的抗菌剂；MEP途径中的所有酶都被认为是新药的潜在靶点 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不可少的 而且人类细胞不包含同源基因。此外，一种天然的抗生素，磷霉素，它可以抑制 途径中的第二种酶(IspC)也抑制革兰氏阴性细菌的生长，我们已经证明 MEP途径(DXS)中第一种酶的抑制剂也能抑制结核分枝杆菌的生长。 然而，很少有实验室能够详细研究这一主题，因为缺乏底物和 适当的高温超导测试。我们以前在这一领域的工作是为了表征MEP通路 来自细菌病原体的酶和开发用于底物的有效合成；我们目前的目标是 识别和开发以前表征的酶的抑制剂，为开发提供潜在的线索 新型抗菌化合物。

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.