Activated Sugars for Drug Discovery

描述(由申请人提供)：这项提议的总体目标是创造一个强大的一锅，体外，多克规模的活性糖合成，用于药物发现和生化研究目的。天然产物糖基化对于发现新的药物化合物是至关重要的。虽然人们已经认识到，许多具有生物活性的天然产物的许多治疗特性都归功于它们的糖基化多样性，但为药物发现创造进一步多样性的方法一直难以开发。其中一个挑战是，创造多样性所需的修饰和稀有糖是无法获得或极难制造的，而且这些糖需要被化学“激活”，以便将它们转移到核心天然产品分子中。然而，已经开发出一种新的技术，其中使用化学酶方法将糖连接到自然产生的苷元上。在这种方法中，需要一种激酶和核苷酸转移酶的组合来形成特定的活性糖，最后利用糖基转移酶将糖转移到合适的苷元核心。这项技术对药物发现很有价值，因为它可以用来生产不同糖基化的天然产物分子库。该项目的第二阶段研究将继续改进这一过程所需的酶，包括设计更大的底物范围。然后，研究将集中在连接最好的酶、ATP/NTP再生、产品回收、工艺放大和创造新的抗生素衍生物上，作为开发技术的示范。与公共健康相关：该项目旨在开发一种用于药物发现目的的制造各种活性糖的方法。这些糖可以用来制造具有新治疗特性的天然产品的衍生品，例如对抗生素耐药性细菌有效的抗生素。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to create a robust one pot, in vitro, multi-gram scale synthesis of activated sugars for drug discovery and biochemical research purposes. Natural product glycosylation is vital for the discovery of new pharmaceutical compounds. While it has been recognized for some time that many biologically active natural products owe much of their therapeutic properties to their glycosylation diversity, methods for creating further diversity for drug discovery have been difficult to develop. One of the challenges is that the modified and rare sugars needed for creating diversity are unavailable or extremely difficult to make, and that the sugars need to be chemically "activated" in order to transfer them to the core natural product molecule. However, new technology has been developed where a chemoenzymatic method is used to attach sugars to natural production aglycons. With this method, a combination of a kinase and nucleotide transferase are needed to form the specific activated sugar and finally a glycosyltransferase is used to transfer the sugar to an appropriate aglycon core. This technology is valuable for drug discovery, as it can be used to produce a library of differentially glycosylated natural product molecules. The Phase II research of this project will continue the improvements in the enzymes needed for this process, including engineering a greater substrate range. The research will then focus on coupling the best enzymes, ATP/NTP regeneration, product recovery, process scale-up and creating novel antibiotic derivatives as a demonstration of the developed technology. PUBLIC HEALTH RELEVANCE: This project is aimed toward developing a method for making a variety of activated sugars for drug discovery purposes. These sugars can used to make derivatives of natural products with new therapeutic properties, for example, antibiotics that are effective against antibiotic-resistant bacteria.