IGF::OT::IGF SYNTHESIS OF HIGH-MANNOSE N-GLYCANS

IGF::OT::IGF 高甘露糖 N-聚糖的合成

• 批准号: 9154667
• 负责人: QINGFENG PAN
• 金额: $ 100万
• 依托单位: OMICRON BIOCHEMICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-18 至 2017-09-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9154667
• 关键词: Address; Bacterial Infections; Binding; Biological Assay; Biological Process; Capillary Electrophoresis; Cell surface; Cells; Chemicals; Commercial Sectors; Communities; Contractor; Development; Epitopes; Glycoproteins; High Pressure Liquid Chromatography; Human; Immune response; Indium-111; Investigation; Knowledge; Libraries; Lipids; Mannose; Mass Spectrum Analysis; Methods; Molecular; Monosaccharides; Oligosaccharides; Outcome; Parents; Phase; Polysaccharides; Positioning Attribute; Preparation; Process; Proteins; Protocols documentation; Research Personnel; Resolution; Scheme; Services; Shipping; Ships; Specificity; Structure; Time; Translations; Work; carbohydrate binding protein; chemical synthesis; commercialization; enzyme substrate; experience; man; polypeptide; screening

Cell-surface oligosaccharides appended to proteins and lipids are key binding epitopes in many critical biological processes, including bacterial infection, cell development and the immune response. Understanding these processes at the molecular level requires access to a wide range of oligosaccharides of known structure to support investigations of enzyme substrate binding and specificity, to screen for carbohydrate binding proteins, and to develop assays for enzymic activity. These oligosaccharides are currently unavailable in the commercial sector, thus impeding progress in this field. To address this deficiency, the proposed project focuses on the synthesis of oligosaccharides derived from the parent high-mannose oligosaccharide (14-mer) that is transferred en-bloc to polypeptides during translation. During Phase I of this• project, a "bottom-up" synthetic approach instead of a "top-down" approach to overcome the limitations of the latter was pursued. All five 0-glycosidic linkages found in naturally occurring high-mannose N-glycans of human glycoproteins were constructed, leading to the preparation of thirty-two (32) oligosaccharides (3- to 6-mers) using a generalized chemical approach. The Phase I project developed robust synthetic protocols, and the knowledge, experience and technical know-how gained during Phase I will help extend the chemical space to over 108 compounds in pending Phase II work. The entire chemical space of nested oligosaccharide fragments containing six or fewer monosaccharides (GlcNAc and Man residues), and nested fragments containing up to nine Man residues, will be covered. Methods will be developed to array this extensive library to support anticipated screening efforts by investigators in the glycoscience community.The Contractor shall provide services for the following: Seventy-six (76) high mannose oligosaccharides ranging in size from tri- to nona-saccharides shall be prepared by chemical synthesis. Each oligosaccharide shall be prepared in 0.2 mg or greater quantities and in high purity, the latter determined by NMR, high resolution mass spectrometry, high-pressure liquid chromatography and/or capillary electrophoresis. The 24-month Phase II project period shall provide the opportunity to significantly expand the high-mannose N-glycan product line and to position the contractor for commercialization of these new glycoscience products. The Phase II synthetic work shall be divided into four (4) project periods, each with specific outcomes in terms of final oligosaccharide products synthesized, characterized, and shipped to the NCI.

在许多关键的生物学过程中，附加到蛋白质的细胞表面寡糖是关键的结合表位，细胞发育和已知结构的免疫反应。酶活性目前在商业部门中，因此诱使绳索项目集中于源自母体高甘露糖寡糖（14-mer）的合成寡糖。 •构建了一种“自下而上”的合成方法，而不是一种克服后者的“上下”方法。到6年左右）I期项目开发了强大的合成协议，经验和技术知识，在第一阶段获得了如何将化学空间扩展到108以上 待处理的II期的化合物。承包商的调查人员应为以下六（76）寡糖提供服务，从三到nona-sacharides范围应通过化学综合进行准备。由NMR确定，高压灯压24个月II期项目Puriod应为显着扩展的扩张扩展扩张的扩张扩展扩大扩张，扩大扩展扩张，扩张扩张扩张，扩大扩展扩大，扩张扩张扩张，扩张扩张扩张，扩大了扩张的扩展，从而扩展了扩张的扩展，并扩大了繁殖膨胀。或毛细管和/或毛细管和/或毛细管高甘露糖N-聚糖prition缝制缝制的缝制性糖性产品。最终寡糖产品的术语合成，表征和发货 到NCI。