In vitro glycorandomization of natural products

天然产物的体外糖随机化

• 批准号: 8392846
• 负责人: Jon Scott Thorson
• 金额: $ 36.58万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-03 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8392846
• 关键词: Amino Acids; Anti-Bacterial Agents; Anti-Infective Agents; Antiviral Agents; Automobile Driving; Biological; Biological Assay; Biological Factors; Bufadienolides; Cardenolides; Catalysis; Cell Proliferation; Chemistry; Ciprofloxacin; Collaborations; Communities; Complement; Complex; Daptomycin; Development; Drug Kinetics; Enzymes; Equilibrium; Evaluation; Evolution; Generations; Glucose; Glycobiology; Glycosides; Goals; In Vitro; Investments; Knowledge; Legal patent; Libraries; Life; Macrolides; Methods; Minocycline; Modeling; Monosaccharides; Mutagenesis; National Institute of Allergy and Infectious Disease; Novobiocin; Nucleotides; Pharmacodynamics; Phase; Phosphotransferases; Positioning Attribute; Predisposition; Production; Pyrimidine; Reaction; Reagent; Screening procedure; Sirolimus; Source; Specificity; Structural Models; Structure; Structure-Activity Relationship; System; Thymidine; Toxic effect; Uridine; Ursidae Family; Variant; Vertebral column; Work; analog; anticancer activity; base; bufadienolide; catalyst; design; design and construction; directed evolution; drug development; drug discovery; functional group; glycosylation; glycosyltransferase; innovation; inorganic phosphate; liquid chromatography mass spectrometry; member; next generation; novel; pathogen; pre-clinical; progenitor; programs; research study; scaffold; small molecule; success; sugar; sugar nucleotide; tool; user-friendly

DESCRIPTION (provided by applicant): The attachment of sugars to naturally-occurring and/or synthetic small molecules can dramatically influence the corresponding mechanism, pharmacodynamics, pharmacokinetics and even patent life of the parental structure. Yet, due to the technical challenges associated with conventional glycosylation strategies, the application of glycosylation in the context of drug discovery and/or development remains underexplored. An overarching aim of this program has been to develop simple and user-friendly chemoenzymatic methods for the differential glycosylation of target scaffolds to ultimately enable such drug discovery/development exploration. Toward this goal, the first phase of this study (years 1-5) led to the key proof of concept for chemoenzymatic glycorandomization (a one pot, three enzyme strategy capable of activating and attaching free monosaccharides to complex natural product scaffolds) and also provided fundamental information regarding two critical, but poorly understood, enzyme classes (anomeric sugar kinases and sugar-1-phosphate nucleotidylyltransferases). The second phase of this project (years 6-10) dramatically expanded the attempted application of this first generation system toward a range of diverse scaffolds, leading to success in many cases but also exposing key limitations of the platform. Importantly, the work conducted during the second phase also led to fundamental new knowledge regarding glycosyltransferase (GT)-catalyzed reactions that serves as the basis from which to launch the third phase of this study. Specifically, the proposed third phase of this study (years 11-15) takes advantage of our recent abilities to evolve highly permissive GTs and also drive GT-catalyzed reactions in reverse to enable next generation single enzyme or dual enzyme transglycosylation strategies for differential glycosylation of an array of structurally-diverse scaffolds (including natural product-based or synthetic, glycosylated or non-glycosylated, parental scaffolds). We propose to take full advantage of the current state of the art to: i) narrow the gaps of knowledge in understanding functional GT structure-activity-relationships; ii) specifically develop a range o catalysts for the production of novel sugar nucleotides (anticipated to be of broad use to the glycobiology community); iii) develop a range of catalysts for the differential glycosylation of a key set of structurally diverse anti-infective and anticancer scaffolds; and iv)exploit the corresponding differentially glycosylated scaffolds as a novel source for the discovery of new antiinfective and anticancer leads. PUBLIC HEALTH RELEVANCE: We propose to develop simple and user-friendly chemoenzymatic methods for the differential glycosylation of complex target scaffolds to enable glycoconjugation as a tool for drug discovery and development. The models selected as part of the current study are anticipated to hold particularly high potential for the discovery of new antibacterial, antiviral and anticancer leads.

描述（由申请人提供）：糖与天然存在和/或合成小分子的连接可以显着影响相应的机制、药效学、药代动力学，甚至母体结构的专利寿命。然而，由于与传统糖基化策略相关的技术挑战，糖基化在药物发现和/或开发中的应用仍未得到充分探索。该计划的首要目标是开发简单且用户友好的化学酶方法，用于目标支架的差异糖基化，以最终实现此类药物发现/开发探索。为了实现这一目标，本研究的第一阶段（第 1-5 年）得出了化学酶糖随机化的关键概念证明（一锅三酶策略，能够激活游离单糖并将其附着到复杂的天然产物支架上），并且还提供了有关两种关键但知之甚少的酶类（异头糖激酶和糖 1-磷酸核苷酸转移酶）的基本信息。该项目的第二阶段（第 6-10 年）极大地将第一代系统的尝试应用扩展到一系列不同的支架，在许多情况下取得了成功，但也暴露了该平台的关键局限性。重要的是，第二阶段进行的工作还带来了有关糖基转移酶（GT）催化反应的基本新知识，这是启动本研究第三阶段的基础。具体来说，本研究拟议的第三阶段（11-15 年）需要 利用我们最近进化出高度许可的 GT 的能力，并反向驱动 GT 催化反应，以实现下一代单酶或双酶转糖基化策略，用于一系列结构多样的支架（包括基于天然产物的或合成的、糖基化或非糖基化，亲本支架）。我们建议充分利用当前的最新技术来：i）缩小理解功能 GT 结构-活动-关系方面的知识差距； ii) 专门开发一系列用于生产新型糖核苷酸的催化剂（预计将广泛用于糖生物学界）； iii) 开发一系列催化剂，用于一组关键的结构多样的抗感染和抗癌支架的差异糖基化； iv)利用相应的差异糖基化支架作为发现新的抗感染和抗癌药物的新来源。 公共健康相关性：我们建议开发简单且用户友好的化学酶方法，用于复杂目标支架的差异糖基化，使糖缀合成为药物发现和开发的工具。作为当前研究的一部分选择的模型预计在发现新的抗菌、抗病毒和抗癌先导化合物方面具有特别高的潜力。