Sugar Building Blocks and Automated Synthesis of Biomedically-Relevant Glycans

 描述（申请人提供）：该项目的总体目标是响应标题为“用于合成生物医学相关碳水化合物（U 01）的简易方法和技术”的RFA-RM-14-015，（I）开拓更有效的化学方法以获得一组差异保护的碳水化合物结构单元的核心--目前是寡糖化学结构的主要瓶颈，（II）实现容易再现的自动化溶液相合成方法，以将这些结构单元连接成聚糖，以及（III）建立自动化纯化方案和分析方法，以确保最终产品的纯度。由于自动化方案比手动程序需要更详细的变量说明，因此此类方案应有助于在自动化和手动合成之间对这些方法进行简单的调整，以建立用于合成一组涉及人类健康和疾病的关键聚糖的标准操作程序。为了实现这些目标，我们将追求五个具体目标：1）优化方法以简化通过单糖的金属催化衍生化合成碳水化合物结构单元; 2）开发利用上述方法合成鼠李糖的路线-基于糖的结构单元，这将导致自动化的可重复的方法来合成与人类病原体假单胞菌的脂多糖核心相关的低聚鼠李糖化合物铜绿; 3）使用上述和其它方法开发半乳糖、岩藻糖和葡糖胺结构单元的合成路线，其将使得能够用可再现的平行自动化溶液相方法合成人乳寡糖的小型文库; 4）开发使用上述和其他方法制备葡糖胺和甘露糖结构单元的合成路线，这将导致可再现的并行自动化解决方案-阶段方法来合成目前在公开可用的微阵列集合中不可用的人蛋白质相关的高级甘露糖型N-聚糖结构;和5）开发合成路线以制备另外的结构单元，然后提供可重复的平行自动化溶液相方法来合成人蛋白质相关的O-质谱标准所需的聚糖结构和相关结构目前在微阵列集合中也不可用。

 DESCRIPTION (provided by applicant): The overall goals of this project, in response to RFA-RM-14-015 titled "Facile Methods and Technologies for Synthesis of Biomedically Relevant Carbohydrates (U01)" are (I) to pioneer more efficient chemical methods to access a core set of differentially protected carbohydrate building blocks-currently a major bottleneck in the chemical construction of oligosaccharides, (II) to achieve readily reproducible, automated solution-phase synthetic methods to connect these building blocks into glycans and (III) to establish automated purification protocols and analysis methods to ensure purity of final products. Because automated protocols require greater specification of variables than manual procedures, such protocols should aid in the facile adaptation of these methods between automated and manual syntheses to set up standard operating procedures for the synthesis of a set of key glycans implicated in human health and disease. To achieve these objectives, five specific aims will be pursued: 1) to optimize methods to simplify the synthesis of carbohydrate building blocks by a metal-catalyzed derivatization of monosaccharides; 2) to develop synthetic routes using the above methods to rhamnose-based sugar building blocks that will result in automated reproducible methods to synthesize oligorhamnose compounds associated with the lipopolysaccharide core of the human pathogen Pseudomonas aeruginosa; 3) to develop synthetic routes using the above and other methods to galactose, fucose, and glucosamine building blocks that will enable reproducible parallel automated solution-phase methods to synthesize small libraries of human milk oligosaccharides; 4) to develop synthetic routes using the above and other methods to make glucosamine and mannose building blocks that will lead to reproducible parallel automated solution-phase methods to synthesize human protein-associated higher mannose-type N-glycan structures currently unavailable in publicly available microarray collections; and 5) to develop synthetic routes to make additional building blocks to then afford reproducible parallel automated solution-phase methods to synthesize human protein-associated O-glycan structures needed for mass spectrometry standards and related structures that are also currently unavailable in microarray collections.