HTS on Riboflavin and Terpene Biosynthetic Enzymes(RMI)

描述(申请人提供)：上个世纪的医学进步极大地降低了传染病造成的发病率和死亡率。另一方面，细菌、真菌、原生动物和病毒感染仍然是发展中国家最重要的死亡因素。即使在发展中国家，对目前使用的所有抗感染药物的抗药性的迅速发展也导致了开发用于控制传染病的新型化疗药物的紧迫性。最近发现的萜类生物合成的非甲氧戊酸途径的酶在革兰氏阴性细菌和疟原虫中是必不可少的，该途径已被确定为疟疾的治疗靶点。同样，已知维生素B2生物合成途径的蛋白质在革兰氏阴性细菌和酵母中是必不可少的。我们正计划利用我们在上述代谢途径方面的专业知识，作为开发适用于机器人处理的稳健和快速的检测方法的基础。具体地说，将开发针对非甲氧戊酸途径的4种酶和核黄素途径的3种分析方法。所有分析都将设计用于光度监测，并将适用于筛选包含数十万种化合物的大型图书馆。根据我们以前的工作，我们已经可以肯定地说，只要付出可以接受的努力，就可以获得所需的试剂。这些测试计划用于堪萨斯大学高通量筛选实验室与教授合作的100,000个药物类化合物化合物库的筛选。库什曼，乔治和叶。将开发其他化验方法，以快速验证在这些筛查中获得的命中结果。与库什曼教授合作的后续工作将针对通过药物化学方法将候选化合物开发成实际的抗感染药物。

DESCRIPTION (provided by applicant): Medical progress in the last century has dramatically reduced the morbidity and mortality caused by infectious diseases. On the other hand, infections by bacteria, fungi, protozoa and viruses continue to be the most important factor of mortality in developing countries. Even in developing countries, the rapidly progressing development of resistances against all presently used anti-infective agents generates this state of urgency for the development of novel chemotherapeutic agents for the control of infectious diseases. The enzymes of the recently discovered non-mevalonate pathway of terpene biosynthesis are known to be essential in gram negative bacteria and in Plasmodium spp., and the pathway has been established as a therapeutic target for malaria. Similarly, the proteins of the vitamin B2 biosynthetic pathway are known to be essential in gram negative bacteria and in yeasts. We are planning to use our expertise in the metabolic pathways mentioned above as the basis for the development of robust and rapid assay methods that are suitable for robotic processing. Specifically, assays will be developed for 4 enzymes of the non-mevalonate pathway and for 3 assays of the riboflavin pathway. All assays will be designed for photometric monitoring and will be suitable for the screening of large libraries comprising hundreds of thousands of chemical compounds. Based on our prior work, we can already say with certainty that the required reagents can be procured with an acceptable effort. These assays are planned to be used for the screening of a 100,000 compound library of drug like compounds at the High Throughput Screening Laboratory at the University of Kansas in cooperation with Profs. Cushman, George and Ye. Additional assay methods will be developed for rapid verification of hits obtained in these screens. Follow-up work in cooperation with Prof. Cushman will be directed at the development of candidate compounds into actual anti-infective drugs by medicinal chemistry methods.