Sugar Building Blocks and Automated Synthesis of Biomedically-Relevant Glycans

 描述(申请人提供)：本项目的总体目标是响应题为“合成生物医学相关碳水化合物(U01)的简易方法和技术”的RFA-RM-14-015，其总体目标是：(1)开创更有效的化学方法来获得一套不同保护的碳水化合物构建块--目前低聚糖化学构建中的一个主要瓶颈--；(2)实现易于重复的自动化溶液相合成方法，将这些构建块连接成多糖；(3)建立自动化的纯化规程和分析方法，以确保最终产品的纯度。因为自动化方案比人工程序需要更多的变量说明，这种方案应该有助于在自动和手动合成之间方便地适应这些方法，以建立合成一组与人类健康和疾病有关的关键多糖的标准操作程序。为了实现这些目标，将追求五个具体目标：1)优化通过金属催化的单糖衍生化来简化碳水化合物构建块合成的方法；2)开发使用上述方法来合成基于鼠李糖的糖构建块的合成路线，这将导致自动化可重复的方法来合成与人类病原体铜绿假单胞菌的脂多糖核心相关的低聚鼠李糖化合物；3)开发使用上述方法和其他方法来合成半乳糖、岩藻糖和氨基葡萄糖构建块的合成路线，这将使可重复使用的并行自动化液相方法能够合成人乳低聚糖小文库；4)利用上述方法和其他方法开发合成路线，以制造氨基葡萄糖和甘露糖构建块，这将导致可重复的平行自动化溶液相方法，以合成目前在公共可获得的微阵列集合中无法获得的人类蛋白质相关的较高甘露糖型N-葡聚糖结构；以及5)开发合成路线，以制造更多的构建块，然后提供可重复使用的平行自动化溶液相方法，以合成质谱学标准所需的人蛋白相关O-葡聚糖结构和目前在微阵列收集中也无法获得的相关结构。

 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.