Discovery, Design, and Development of Phosphonic Acid Antibiotics

磷酸是一类有效的、但未得到充分利用的生物活性化合物，在治疗人类疾病方面前景广阔。自然界中产生的磷酸盐种类繁多，其中许多具有有用的生物活性。其中包括磷霉素，一种常见的妊娠期急性膀胱炎治疗方法，以及FR900098，一种新的治疗方法(和治愈！)治疗疟疾。其他包括膦硫代三肽 广泛使用的除草剂(Pit)和K-26，一种在治疗高血压方面很有前途的化合物。磷酸盐抑制剂的靶点差异很大，允许它们用于治疗各种疾病，然而，尽管它们有用，但令人惊讶的是，人们对这些天然产物的生物合成知之甚少。正如本提案所概述的那样，我们相信有充分的理由相信：(I)还有更多的天然膦酸产品有待发现(包括具有新的治疗靶点的产品)；(Ii)在未来的几年中，将会发现大量新的生物化学和次生代谢。 (3)可以对目前已知的化合物进行修饰和改进；(4)可以开发出更经济的生产方法。拟议的计划项目通过涉及微生物学、生物化学、化学、代谢工程和结构生物学的多学科研究计划，解决了这些主题中的每一个。我们提出了四个相互交织的研究项目，以发现、设计和开发新的和已知的膦酸类抗生素。 第一个项目涉及利用微生物遗传学和分子生物学发现、鉴定和表征编码磷酸生物合成的基因簇。第二个项目涉及这些生物合成途径的化学和生化表征，包括合成这些含有C-P键的化合物所涉及的几个完全前所未有的酶反应。第三个项目 涉及天然和设计的膦酸生物合成途径的生物工程，以允许过量生产膦酸抗生素。第四个项目的重点是开发和使用质谱学来检测和量化磷酸，并对合成过程中涉及的生化反应进行表征。该项目将设在伊利诺伊大学新的基因组生物学研究所，项目团队将在那里占用一个单独的大型连续实验室。

Phosphonic acids represent a potent, yet underexploited, group of bioactive compounds with great promise in the treatment of human disease. A wide variety of phosphonates are produced in nature and many have useful bioactive properties. Among these are fosfomycin, a common treatment for acute cystitis during pregnancy, and FR900098, a novel treatment (and cure!) for malaria. Others include phosphinothricintripeptide (PIT), a widely used herbicide, and K-26, a compound with great promise in the treatment of high blood pressure. The targets of phosphonate inhibitors vary widely, allowing their use in the treatment of a wide variety of conditions, yet despite their utility, surprisingly little is known regarding the biosynthesis of these natural products. As outlined in this proposal, we believe there is ample reason to believe that (i) additional natural phosphonic acid products remain to be discovered (including ones with novel therapeutic targets), (ii) that a wealth of novel biochemistry and secondary metabolism will be discovered during characterization of phosphonic acid biosynthetic pathways, (iii) that currently known compounds can be modified and improved and (iv) that more economical methods for their production can be developed. The proposed Program Project addresses each of these topics via a multidisciplinary research program involving microbiology, biochemistry, chemistry, metabolic engineering and structural biology. We propose four intertwined research projects to discover, design and develop novel and known phosphonic acid antibiotics. The first project involves discovery, identification and characterization of gene clusters encoding phosphonic acid biosynthesis using microbial genetics and molecular biology. The second project involves the chemical and biochemical characterization of these biosynthetic pathways, including several entirely unprecedented enzymatic reactions involved in synthesis of these C-P bond-containing compounds. The third project involves bioengineering of natural and designed phosphonic acid biosynthetic pathways to allow overproduction of phosphonic acid antibiotics. The fourth project is focused on the development and use of mass-spectrometry to detect and quantify phosphonic acids and to characterize the biochemical reactions involved in their synthesis. The project will be housed in the new Institute for Genomic Biology at the University of Illinois, where the program project team will occupy a single, large contiguous laboratory.