Activated Sugars for Drug Discovery

描述（由申请人提供）：本提案的总体目标是创建一个强大的一锅，体外，多克规模的合成活性糖，用于药物发现和生化研究目的。天然产物糖基化对新药物化合物的发现至关重要。虽然人们已经认识到许多具有生物活性的天然产物的治疗特性在很大程度上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很难开发。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，并且糖需要化学“激活”才能将它们转移到核心的天然产物分子中。然而，一种新的技术已经被开发出来，一种化学酶的方法被用来将糖附着在天然生产的糖基上。在这种方法中，需要激酶和核苷酸转移酶的结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基核心。这项技术对于药物发现是有价值的，因为它可以用来产生一个差异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该工艺所需的酶，包括设计更大的底物范围。然后，研究将集中在最佳酶的偶联，ATP/NTP再生，产品回收，工艺扩大和创造新的抗生素衍生物，作为已开发技术的示范。公共卫生相关性：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物的衍生物，例如，对耐抗生素细菌有效的抗生素。

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.