Self-Assembled Encapsulated Enzymes for Pharmaceutical Synthesis

用于药物合成的自组装封装酶

• 批准号: 8394775
• 负责人: JASON DAVID FIEDLER
• 金额: $ 15.38万
• 依托单位: QAPSULE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8394775
• 关键词: Achievement; Aldehyde-Lyases; Bacteriophages; Benchmarking; Benign; Biological; Businesses; Capsid; Catalysis; Characteristics; Chemicals; Development; Diffuse; Digestion; Disadvantaged; Drug Industry; Encapsulated; Enzymes; Exhibits; Fluorine; Goals; Heating; Immobilized Enzymes; Legal patent; Marketing; Measures; Methods; Modification; Molecular; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Prize; Process; Production; Property; Proteins; Quantitative Evaluations; Reaction; Relative (related person); Research; Solid; Solvents; Staging; Structure; System; Techniques; Technology; Temperature; Testing; Time; Virus; Virus-like particle; Work; analog; case-by-case basis; catalyst; chemical synthesis; cost; cost effective; directed evolution; enzyme structure; enzyme substrate; extreme temperature; improved; interest; man; nanoparticle; operation; particle; protein expression; remediation; success

DESCRIPTION (provided by applicant): With modern requirements for the control of absolute configuration in drug molecules and for their environmentally responsible and cost-effective production, enzymes have become increasingly valuable catalysts for the large-scale synthesis of pharmaceutical intermediates. However, enzymes are sensitive to elevated temperatures and the presence of co-solvents, and are often not optimal in terms of rate, lifetime, product inhibition, and substrate acceptance. The standard ways to ameliorate these problems are to immobilize enzymes on solid supports to improve their batch recyclability, and to improve the properties of enzymes by directed evolution. Both techniques require much time and effort on a case-by-case basis. We have developed a method to incorporate multiple copies of active enzymes inside the very stable protein shell of the bacteriophage QB capsid. These particles can be produced and isolated in large quantities, are extraordinarily stable against denaturation and protease digestion, and impart additional stability to the proteins packaged inside. When enzyme substrates and products can diffuse through the large pores in the capsid structure, the enzymes entrained within can exhibit high catalytic activities. Qapsule Technologies, Inc. proposes to develop such particles for the synthesis of high-value-added pharmaceutical intermediates. This Phase I proposal identifies two proof-of-concept targets - an aldolase for asymmetric catalysis and a fluorinase for the installation of fluorine atoms, both highly prized by the pharmaceutical industry. We will demonstrate the high-yield production of nanoparticles containing multiple copies of these enzymes, determine their catalytic activities, and test their stabilities, activities, lifetime, and process characteristics relative to non-encapsulated analogues. Clear benchmarks for success are identified, the achievement of which will set the stage for Phase II development and the initiation of a business plan to bring these capabilities to market. Our technology for enzymatic production and packaging can be universally applied to almost any enzyme of interest. It therefore adds to efforts of directed evolution and provides a constant platform that will streamline production, use, and workup of biocatalytic engines for large-scale pharmaceutical synthesis. PUBLIC HEALTH RELEVANCE: Enzymes are the catalysts of choice for many transformations used in the synthesis of pharmaceutical intermediates, but are often too unstable for efficient use. We will develop robust protein nanoparticles containing functional enzymes, which stabilize these catalysts and allow for convenient production and processing.

描述（由申请人提供）：随着对药物分子中绝对构型的控制以及对它们的环境负责和成本有效的生产的现代要求，酶已经成为用于大规模合成药物中间体的越来越有价值的催化剂。然而，酶对升高的温度和共溶剂的存在敏感，并且在速率、寿命、产物抑制和底物接受方面通常不是最佳的。改善这些问题的标准方法是将酶固定在固体载体上以改善其批次可回收性，并通过定向进化来改善酶的性质。这两种技术都需要大量的时间和精力在个案的基础上。我们已经开发了一种方法，将多个拷贝的活性酶的非常稳定的蛋白质壳的噬菌体QB衣壳。这些颗粒可以大量生产和分离，对变性和蛋白酶消化非常稳定，并赋予包装在里面的蛋白质额外的稳定性。当酶底物和产物可以通过衣壳结构中的大孔扩散时，夹带在其中的酶可以表现出高催化活性。Qaptek Technologies，Inc.提出开发这种颗粒用于合成高附加值的药物中间体。该第一阶段提案确定了两个概念验证目标-用于不对称催化的醛缩酶和用于安装氟原子的氟化酶，这两个目标都受到了科学家的高度重视。 制药业。我们将展示含有这些酶的多个拷贝的纳米颗粒的高产率生产，确定它们的催化活性，并测试它们的稳定性，活性，寿命和相对于非封装类似物的工艺特性。确定了成功的明确基准，实现这些基准将为第二阶段的发展和启动业务计划奠定基础， 市场我们的酶生产和包装技术可普遍应用于几乎所有感兴趣的酶。因此，它增加了定向进化的努力，并提供了一个恒定的平台，将简化大规模药物合成的生物催化引擎的生产，使用和处理。 公共卫生关系：酶是用于药物中间体合成中的许多转化的催化剂的选择，但通常太不稳定而不能有效使用。我们将开发含有功能性酶的坚固蛋白质纳米颗粒，使这些催化剂稳定并便于生产和加工。