Synthetic Heparan Sulfate: Probing Biosynthesis to Prepare Defined Drugs

合成硫酸乙酰肝素：探索生物合成以制备确定的药物

• 批准号: 8436566
• 负责人: PAUL L DEANGELIS
• 金额: $ 41.95万
• 依托单位: RENSSELAER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-12-15 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8436566
• 关键词: Acetylation; Acids; Anabolism; Binding Proteins; Biochemical; Biochemistry; Biological; Biological Assay; Biological Process; Biology; Blood Coagulation Factor; Bypass; Catabolism; Cell Culture Techniques; Cell Proliferation; Cells; Coagulation Process; Complex; Development; Disaccharides; Disease; Embryonic Development; Engineering; Enzymes; Event; Exhibits; FGF1 gene; Fibroblast Growth Factor; Fibroblast Growth Factor Receptors; Glycoconjugates; Glycoproteins; Goals; Golgi Apparatus; Grant; Growth Factor; Growth Factor Receptors; Growth and Development function; Health; Hemostatic function; Heparan Sulfate Biosynthesis; Heparin; Heparitin Sulfate; Human body; In Vitro; Individual; Inorganic Sulfates; Isomerism; Knock-out; Label; Learning; Length; Libraries; Link; Malignant Neoplasms; Mammals; Maps; Mass Spectrum Analysis; Measures; Mediating; Methodology; Methods; Modeling; Modification; N Domain; Nuclear Magnetic Resonance; Oligosaccharides; Pattern; Pharmaceutical Preparations; Play; Polymers; Polysaccharides; Protein Isoforms; Proteins; Receptor Signaling; Recombinants; Reporting; Role; Route; Series; Signal Transduction; Specificity; Staging; Structure; Structure-Activity Relationship; Surface Plasmon Resonance; System; Testing; Therapeutic; Transgenic Organisms; Unspecified or Sulfate Ion Sulfates; Uridine Diphosphate Sugars; Variant; Vertebral column; amino group; analog; base; biological systems; cell growth; combinatorial; cytokine; design; domain mapping; enzyme activity; epimerase; extracellular; heparanase; heparosan synthase; high throughput screening; improved; in vivo; milligram; next generation; preference; receptor; research study; sugar; sulfated polymer; sulfation; sulfotransferase

DESCRIPTION (provided by applicant): Heparan sulfate (HS), a class of polysaccharides that includes the well-known drug heparin, plays essential roles in cell growth and development. The many structural variants of HS observed in mammals have been hypothesized to differentially modulate growth factor-mediated signaling and regulate hemostasis during health and disease. Our project will explore how the HS structural variation is generated during biosynthesis/catabolism as well as map routes to prepare more defined molecules having greater selectivity and more potent desired bioactivities. A favored hypothesis is that the pattern of HS domains (comprised of both N-sulfo (NS) and N-acetyl (NA) domains) encodes information that modulates HS interaction with proteins like growth factors, cytokines, and clotting factors. Currently, it is very difficult to understand how HS biosynthetic modification an domain placement is controlled and which HS domain structures possess the highest activities in different biological systems. We will apply our newly developed synthetic methodology to answer key questions in the field. A small library of HS polymers having size-defined and placement-defined NS and NA domains will be generated. These HS polymers will then be modified by biosynthetic enzymes, including O-sulfotransferases, and C5-epimerase and/or catabolic enzymes including endo-6- Oendosulfatases and heparanase. A focused combinatorial approach will be used to produce defined HS polymers to test two opposing models of growth factor signaling, a key event in cell proliferation, embryonic development, and cancer. In this project our specific aims are: Aim 1: To chemoenzymatically synthesize a small library of HS polymers having defined NS and NA domains. Aim 2. To analyze the modification patterns generated through the action of various enzymes on the HS polysaccharide library. Aim 3: To characterize the structure/function relationship of HS activity as the co-receptor fibroblast growth factor receptor signaling.

描述（由申请人提供）：硫酸乙酰肝素（HS）是一类多糖，包括众所周知的药物肝素，在细胞生长和发育中起重要作用。已经假设在哺乳动物中观察到的HS的许多结构变体在健康和疾病期间差异地调节生长因子介导的信号传导并调节止血。我们的项目将探索HS结构变异是如何在生物合成/催化过程中产生的，以及绘制路线以制备具有更大选择性和更有效的所需生物活性的更明确的分子。一个有利的假设是， HS结构域（由N-磺基（NS）和N-乙酰基（NA）结构域组成）编码调节HS与蛋白质如生长因子、细胞因子和凝血因子相互作用的信息。目前，很难理解HS生物合成修饰结构域的位置是如何控制的，以及在不同的生物系统中哪些HS结构域结构具有最高的活性。我们将应用我们新开发的合成方法来回答该领域的关键问题。将产生具有尺寸限定和位置限定的NS和NA结构域的HS聚合物的小文库。然后，这些HS聚合物将被生物合成酶修饰，所述生物合成酶包括O-磺基转移酶和C5-差向异构酶和/或分解代谢酶，所述分解代谢酶包括内切-6-O-硫酸酯酶和乙酰肝素酶。一个集中的组合方法将用于生产定义的HS聚合物，以测试两个相反的模型的生长因子信号，在细胞增殖，胚胎发育和癌症的关键事件。在这个项目中，我们的具体目标是：目标1：化学酶法合成一个小的库HS聚合物具有确定的NS和NA结构域。 目标二。分析各种酶对HS多糖库的作用所产生的修饰模式。目的3：研究HS作为成纤维细胞共受体的结构/功能关系 生长因子受体信号传导