Solution-Phase Automated Synthesis of Oligosaccharides

描述（由申请人提供）：碳水化合物的合成仍然是一项复杂的工作，涉及大量的劳动力和大量昂贵的构建模块和试剂。即使一种制造碳水化合物的方法已经被开发出来，生物学家为研究驱动糖组学而要求的各种分子的制造仍然是劳动和材料密集型的。由于很难从天然来源中分离出足够和纯量的碳水化合物，因此缺乏现成的一系列碳水化合物结构，这不仅严重限制了科学的发展，而且严重限制了由此产生的基于碳水化合物的疫苗和基于这种基本认识的诊断方法的发展。拟议项目的长期目标是开发商业定制合成低聚糖，使用稳定的，如果可能的话，晶体构建块与构建块组装的创新自动化解决方案相结合。第一阶段的建议解决了现阶段阻碍这一进程发展的障碍。具体目的是：1)制备一系列n -苯基取代的三氟乙酰咪啶酰卤化物作为糖基活化剂；2)制备了不同的三氟乙酰胺基糖基供体，并对其稳定性和反应性进行了测试；3)这些给体与一系列糖基受体反应形成双糖文库，这是一种基于溶液相的自动化策略，适用于迭代低聚糖合成。在这些研究的结论中，将发现稳定的糖基供体与广泛的糖基受体在液相自动化平台中反应，为按需定制顺序合成低聚糖的标准化碳水化合物构建块铺平道路。

DESCRIPTION (provided by applicant): The synthesis of carbohydrates remains a complex exercise that involves tremendous labor and large amounts of expensive building blocks and reagents. Even once a process for making a carbohydrate has been developed, manufacture of that molecule in all the variants that biologists demand for their studies to drive glycomics remains labor and material intensive. The lack of ready access to a range of carbohydrate structures, given the extreme difficulty in isolating sufficient and pure quantities from natural sources, has been a severe limitation to advancing not only the science but also the resulting carbohydrate based vaccines and diagnostics built on this basic understanding. The long-term goal of the proposed project is to develop the commercial custom synthesis of oligosaccharides using stable and, if possible, crystalline building blocks in association with an innovative new automated solution-phase process for building block assembly. This phase I proposal addresses roadblocks that stand in the way of the development of this process at this stage. The specific aims are 1) the preparation of a range of N-phenyl substituted trifluoroacetimidoyl halides as glycosyl activating reagents; 2) the preparation of different trifluoroacetimidoyl based glycosyl donors with tests of their stability and reactivity; and 3) the reaction of these donors with a range of glycosyl acceptors to form disaccharide libraries in a solution-phase-based automation strategy amenable to iterative oligosaccharide synthesis. At the conclusion of these studies, stable glycosyl donors reactive with a wide range of glycosyl acceptors in a solution phase automation platform will have been discovered to pave the way for a standardized set of carbohydrate building blocks for on-demand custom-order oligosaccharide synthesis. PUBLIC HEALTH RELEVANCE: Carbohydrates are increasingly recognized as fundamental molecular constituents in a wide range of cellular processes of medical importance, including infectious disease responses (fungal, e.g candidiasis; viral, e.g. influenza; and bacterial, e.g. Staphylococcus aureus), cancer progression, and congenital metabolic disorders (e.g. mucopolysaccharidosis). This proposal will develop methods for the first rapid automated assembly of oligosaccharides in solution for use in medical diagnostics and basic science studies.