Chemo-enzymatic synthesis of nucleotide sugar libraries and therapy-relevant derivatives

核苷酸糖库和治疗相关衍生物的化学酶合成

• 批准号: 10225322
• 负责人: Wanyi Guan
• 金额: $ 34.19万
• 依托单位: CHEMILY, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10225322
• 关键词: Binding; Carbohydrates; Cell Differentiation process; Cell surface; Cells; Chemicals; Communities; Coupled; Coupling; DNA; Development; Disease; Drug Delivery Systems; Fluorides; Future; Generations; Glycoconjugates; Goals; Health; Human; Immune response; In Situ; Individual; Libraries; Link; Mediating; Medical Research; Methods; Modification; Nature; Nucleotides; Phase; Play; Polysaccharides; Positioning Attribute; Process; Production; Protein-Carbohydrate Interaction; Reaction; Reagent; Role; Small Business Innovation Research Grant; Structure; System; Technology; Therapeutic; Tissues; Transferase; Uridine Diphosphate Sugars; Variant; base; cancer cell; cell type; chemical synthesis; commercialization; enzyme pathway; experimental study; functional group; glycosylation; glycosyltransferase; individualized medicine; innovation; man; nucleotide metabolism; personalized medicine; scale up; sugar; sugar nucleotide; targeted treatment

Glycans and glycoconjugates are widespread in nature and play pivotal roles in cell-cell recognition, cell differentiation, and immunological responses. Most glycans and glycosylation reactions require nucleotide sugars as their essential building blocks. Therefore, the commercial availability of nucleotide sugars will determine future opportunities for carbohydrate-based innovations, just as the availability of nucleotides was a prerequisite for today’s $6.1 billion PCR market of DNA-based technologies. Due to glycoscience’s reliance on nucleotide sugars, there is an urgent need for a production pipeline to provide these essential reagents. The goal of our “Chemo-enzymatic synthesis of nucleotide sugar libraries and therapy-relevant derivatives” Fast-Track project is to meet this need and commercialize a stable, affordable and diverse supply of nucleotide sugars to drive advances in glycoscience. It is our long-term objective to bring 11 nucleotide sugars, 26 nucleotide derivatives, and the associated processes to market. In Phase I Aim 1, we will enzymatically synthesize GDP-L-Fuc, GDP-Man, CMP-Neu5Ac, CMP-Neu5Gc, and 7 UDP sugars: Glc, Gal, GlcNAc, GalNAc, GlcA, GalA, and Xyl. In Aim 2, we will create optimized in situ generation systems for the UDP-, GDP-, and CMP-sugars to be coupled with transferase reactions. Our methods will utilize de novo or salvage pathway enzymes and be scaled up to 1 gram production reactions. In Phase II Aim 1, we will chemically modify the nucleotide sugars to form 26 new derivatives containing azido, fluoride, deoxy or methoxy functional groups at various positions on CMP-Neu5Ac, GDP-L-Fuc, and UDP-GlcNAc/GalNAc. In Aim 2, we will further optimize all 37 production platforms for commercialization. Our methods will combine chemical synthesis with de novo and salvage pathway enzyme reactions to produce the most efficient products. The health relevance of this project is linked to developments in glycan technology, due the fundamental nature of nucleotide sugars as building blocks for glycoconjugates and polysaccharides. A stable supply of nucleotide sugars and their derivatives will facilitate experiments that advance our understanding of cell surface markers, cell recognition, and protein-carbohydrate interactions. In turn, this cell specific information has the potential to impact individualized medicine, drug-delivery, and cell targeting therapies.

聚糖和复合糖在自然界中广泛存在，在细胞间发挥着关键作用 识别、细胞分化和免疫反应。大多数聚糖和糖基化反应 需要核苷酸糖作为其基本构件。因此，商业可用性 核苷酸糖将决定未来基于碳水化合物的创新机会，就像 核苷酸的可用性是当今价值 61 亿美元的基于 DNA 的 PCR 市场的先决条件 技术。由于糖科学对核苷酸糖的依赖，迫切需要一种 生产管道提供这些必需的试剂。 我们的“核苷酸糖库的化学酶合成和治疗相关的目标” 衍生品”Fast-Track项目就是为了满足这一需求，并将稳定、实惠且多样化的产品商业化 供应核苷酸糖以推动糖科学的进步。我们的长期目标是使 11 核苷酸糖、26 种核苷酸衍生物以及相关工艺推向市场。在第一阶段目标 1 中， 我们将酶法合成 GDP-L-Fuc、GDP-Man、CMP-Neu5Ac、CMP-Neu5Gc 和 7 UDP 糖：Glc、Gal、GlcNAc、GalNAc、GlcA、GalA 和 Xyl。在目标 2 中，我们将创建原位优化的 UDP-、GDP-和 CMP-糖的生成系统与转移酶反应相结合。我们的 方法将利用从头或挽救途径酶并扩大至 1 克产量 反应。在第二阶段目标1中，我们将对核苷酸糖进行化学修饰，形成26种新的衍生物 CMP-Neu5Ac 上的不同位置含有叠氮基、氟化物、脱氧或甲氧基官能团， GDP-L-Fuc 和 UDP-GlcNAc/GalNAc。在目标2中，我们将进一步优化所有37个生产平台 商业化。我们的方法将化学合成与从头合成和回收途径结合起来 酶反应可产生最有效的产品。 该项目的健康相关性与聚糖技术的发展有关，因为 核苷酸糖作为复合糖和多糖的基本性质。一个 核苷酸糖及其衍生物的稳定供应将有助于推进我们的实验 了解细胞表面标记、细胞识别和蛋白质-碳水化合物相互作用。反过来， 这种细胞特异性信息有可能影响个体化医疗、药物输送和细胞 靶向治疗。