Activated Sugars for Drug Discovery

描述（由申请人提供）：本提案的总体目标是创建用于药物发现和生物化学研究目的的活性糖的稳健的一锅法、体外、多克规模合成。天然产物糖基化对于发现新的药物化合物至关重要。虽然一段时间以来人们已经认识到，许多生物活性天然产物的治疗特性大部分归功于其糖基化多样性，但难以开发用于药物发现的进一步多样性的方法。其中一个挑战是，创造多样性所需的修饰和稀有糖是不可用的或极难制造的，并且糖需要被化学“活化”，以便将它们转移到核心天然产物分子中。然而，已经开发了新技术，其中使用化学酶法将糖连接到天然产物糖苷配基。用这种方法，需要激酶和核苷酸转移酶的组合来形成特定的活化糖，最后使用糖基转移酶将糖转移到适当的糖苷配基核心。该技术对于药物发现是有价值的，因为它可以用于产生差异糖基化的天然产物分子的文库。该项目的第二阶段研究将继续改进该过程所需的酶，包括设计更大的底物范围。然后，研究将集中在耦合最好的酶，ATP/NTP再生，产品回收，工艺放大和创造新的抗生素衍生物作为开发技术的示范。公共卫生相关性：该项目旨在开发用于药物发现目的的各种活性糖的制造方法。这些糖可以用来制造具有新的治疗特性的天然产物衍生物，例如，对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.