Synthesis of the library of heparan sulfate hexasaccharide mimetics

项目总结： 硫酸乙酰肝素(HS)在许多重要的生物学事件中发挥作用，包括癌症转移，抗肿瘤转移，抗肿瘤作用。 凝血和阿尔茨海默病的发展。HS结构在性质上是异质的，范围很广 硫酸盐化模式和主干结构。已知HS与其生物伴侣的相互作用是 受HS细微结构的显著影响。因此，为了彻底理解结构-活性 关于HS的关系，提供大型HS结构库是至关重要的。然而，HS的合成 由于HS结构的复杂性，寡糖库仍然是一个巨大的挑战。要克服 这一瓶颈，通过创新的产业界/学术界合作，葡聚糖治疗公司将开发一种HS 如具有不同和明确分子结构的六糖文库。一种新型的六糖 将通过HS双糖的头尾连接来设计模拟分子，从而大大简化整体 能够制备大量化合物用于文库合成的合成方法。 在目标1中，将开发基于溶液相的方法来合成HS类六糖 模仿当地的HS。强大的连接体化学将被设计成以头尾相接的方式连接双糖模块 时尚。在目标2中，将开发方法来方便地合成关键的双糖构建块。至 涵盖了自然界中遇到的含有2-O、6-O和N-硫酸盐的不同HS序列，16种二糖是 需要的。而不是从相应的单糖从头开始，一种新的发散合成 将制定战略，其中16种双糖将来自两种关键的高级双糖 积木。这一过程减少了生产积木所需的合成步骤总数 增加了约50%。在目标3中，将开发自动化化学以实现类似HS的机器辅助合成 六糖模拟文库。每种化合物(纯度98%)的结构将通过1D/2D核磁共振进行确认 和高分辨率质谱仪。所有模拟都将在还原端使用链接器来实现功能化 使生物偶联和微阵列生产变得容易。 在第一阶段研究中，两种关键的战略保护的高级双糖中间体(各500毫克)， 并将生产6个新的六糖模拟物(每个2 mg)。此外，自动化方法将是 开发用于合成两种六糖类化合物(各2毫克)。在第二阶段，合成将是 扩大规模以生产300个HS类六糖(每个2毫克)的文库。 从这个项目中组装的广泛的HS六糖模拟物将是最大的 和最全面的HS模仿库。这些产品的商业供应将 极大地促进了对HS迷人的生物学功能的理解以及对 以HS为基础的新型疗法的发展。

Project summary: Heparan sulfate (HS) play roles in many important biological events, including cancer metastasis, anti- coagulation, and Alzheimer’s disease development. HS structures in nature are heterogeneous with a wide range sulfation patterns and backbone structures. The interactions of HS with their biological partners are known to be significantly impacted by the fine structures of HS. Thus, for a thorough understanding of the structure-activity relationships of HS, it is critical that large libraries of HS structures are available. However, synthesis of HS oligosaccharide libraries remains a tremendous challenge due to the complexity of HS structures. To overcome this bottleneck, through an innovative industry/academia collaboration, Glycan Therapeutics will develop a HS like hexasaccharide library with diverse and well-defined molecular structures. The novel hexasaccharide mimetics will be designed through head-to-tail linkage of HS disaccharides, thus greatly simplifying the overall synthetic process enabling the preparation of a large number of compounds for library synthesis. In aim 1, solution phase based methodologies will be developed to synthesize HS like hexasaccharides mimicking native HS. Robust linker chemistry will be designed to link the disaccharide modules in a head-to-tail fashion. In aim 2, methods will be developed for expedient synthesis of the key disaccharide building blocks. To cover the diverse HS sequences bearing 2-O, 6-O and N-sulfation encountered in nature, 16 disaccharides are needed. Rather than starting from the corresponding monosaccharides de novo, a new divergent synthetic strategy will be developed where the 16 disaccharides will be derived from two key advanced disaccharide building blocks. This process reduces the total number of synthetic steps needed to produce the building blocks by ~50%. In aim 3, automation chemistry will be developed to enable machine aided synthesis of the HS like hexasaccharide mimetic library. The structure of each compound (>98% purity) will be confirmed by 1D/2D NMR and high-resolution mass spectrometry. All mimetics will be functionalized with a linker at the reducing end to enable facile bioconjugation and microarray production. In Phase I studies, two key strategically protected advanced disaccharide intermediates (500 mg each), and 6 new hexasaccharide mimetics (2 mg each) will be produced. In addition, automated method will be developed to synthesize two hexasaccharide like compounds (2 mg each). In Phase II, the synthesis will be scaled up to produce a library of 300 HS like hexasaccharides (2 mg each). The extensive HS hexasaccharide mimetics that will be assembled from this project will be the largest and the most comprehensive HS mimic library available. The commercial availability of these products would greatly accelerate research on the understanding of the fascinating biological functions of HS, as well as the development of novel HS-based therapeutics.