Self-Assembled Encapsulated Enzymes for Pharmaceutical Synthesis

描述(由申请人提供)：随着对药物分子绝对构型的控制以及对环境负责和成本效益高的生产的现代要求，酶已成为大规模合成药物中间体的越来越有价值的催化剂。然而，酶对高温和共溶剂的存在很敏感，在速率、寿命、产物抑制和底物接受度方面往往不是最优的。改善这些问题的标准方法是将酶固定在固体载体上以提高其批量回收利用能力，并通过定向进化来改善酶的性质。根据具体情况，这两种技术都需要花费大量的时间和精力。我们已经开发出一种方法，将活性酶的多个拷贝整合到噬菌体QB衣壳的非常稳定的蛋白质外壳中。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，并为包装在其中的蛋白质提供了额外的稳定性。当酶底物和产物可以通过衣壳结构中的大孔隙扩散时，携带在其中的酶可以显示出高催化活性。Qapsule Technologies，Inc.建议开发这种颗粒，用于合成高附加值的医药中间体。这个第一阶段的提案确定了两个概念验证目标-一个用于不对称催化的醛缩酶和一个用于安装氟原子的氟化酶，这两个目标都受到了 制药业。我们将展示包含这些酶的多个副本的纳米颗粒的高产量生产，确定它们的催化活性，并测试它们相对于非胶囊类似物的稳定性、活性、寿命和工艺特征。确定了明确的成功基准，这些基准的实现将为第二阶段的开发和启动业务计划奠定基础，以实现这些能力 市场。我们的酶生产和包装技术可以普遍适用于几乎任何感兴趣的酶。因此，它增加了定向进化的努力，并提供了一个持续的平台，将简化用于大规模药物合成的生物催化发动机的生产、使用和工作。 与公众健康相关：酶是用于合成药物中间体的许多转化的首选催化剂，但往往太不稳定，无法有效使用。我们将开发包含功能性酶的强大的蛋白质纳米颗粒，这种纳米颗粒可以稳定这些催化剂，并允许方便的生产和加工。

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.