STTR Phase I: Enzymatic Synthesis of Chiral Cyclopropanes for Pharmaceutical Drug Synthesis and Agricultural Crop Protection Applications

这个小企业创新研究第一阶段项目的更广泛的影响/商业潜力是开发新的、突破性的酶催化反应，可应用于药品和作物保护剂的生产。通过建立一种广泛适用的生物催化替代方法来生产手性环丙烷这类重要的化合物，Provivi将创造更安全、更清洁、更低成本的合成路线。在大多数情况下，这种新的生物催化反应的应用将减少步骤的数量，并降低合成这些关键组成部分所需的资本投资。在这项研究中开发的新酶技术将改善现有药物和目前药物开发管道中化合物的合成。进一步的应用设想在生产新的作物保护剂。正在开发的酶具有使用现代分子生物学方法对每个特定目标产品进行优化的优点。此外，在水条件下进行反应将减少对有机溶剂的需求，提高过程的可持续性。用更高效和可持续的酶催化步骤取代现有的化学路线将降低成本并提高用于药物合成的许多先进药物中间体的纯度。本一期研究项目的技术目标是演示新型酶促环丙烷化反应在多种商业原料药生产中的应用。手性环丙烷是在许多药物和作物保护化合物中发现的关键亚结构。用于合成这些化合物的含环丙烷的构建块包含至少一个，通常不止一个手性中心。由于生物活性通常需要具有单一的立体异构体，因此不断寻求实现高立体选择性的化学方法。对于环丙烷化反应，现有的方法通常依赖于过渡金属催化剂，如含铑手性配体。与当代化学方法相比，生物催化方法具有明显的优势，因为它将避免使用稀有、昂贵的金属和昂贵的辅助配体来进行这些类型的反应。也要避免高温和恶劣条件。在这项研究中，高通量筛选将用于鉴定以更高的速率和更高的立体选择性催化所需环丙烷化反应的改进变体。通过开发一套扩展的环丙烷化生物催化剂，使其能够作用于更广泛的起始材料，这种新型酶促反应的范围和效用将得到增加。

The broader impact/commercial potential of this Small Business Innovation Research Phase I project is to develop novel, breakthrough enzyme-catalyzed reactions that can be applied to the production of pharmaceuticals and crop protection agents. By establishing a broadly applicable biocatalytic alternative to produce an important class of compounds called chiral cyclopropanes, Provivi will create safer, cleaner, and lower cost synthetic routes. In most cases the application of this new biocatalytic reaction will reduce the number of steps and lower the required capital investment for the synthesis of these key building blocks. The new enzyme technology being developed in this research will improve the synthesis of both existing drugs and compounds in current drug development pipelines. Further applications are envisioned in the production of new crop protection agents. The enzymes being developed have the advantage of being optimizable for each specific target product using modern molecular biology methods. Furthermore, performing the reactions in aqueous conditions will reduce the need for organic solvents, improving the sustainablility of the processes. Replacing existing chemical routes with the more efficient and sustainable enzyme-catalyzed steps will reduce the cost and improve the purity of many advanced pharmaceutical intermediates used in drug synthesis.The technical objectives of this Phase I research project are to demonstrate the application of the novel enzymatic cyclopropanation reaction to the production of a variety of commercial drug substances. Chiral cyclopropanes are key substructures found in a number of pharmaceutical and crop protection compounds. The cyclopropane-containing building blocks used in the synthesis of these compounds contain at least one, and often more than one, chiral center. Since biological activity typically requires having a single stereoisomer, chemical methods that achieve high stereoselectivity are continually sought. For cyclopropanation reactions, the existing methods typically rely on transition-metal catalysts such as rhodium bearing chiral ligands. The biocatalytic method offers clear advantages over the contemporary chemistry in that it will circumvent the use of rare, expensive metals and costly auxiliary ligands for these types of reactions. High temperatures and harsh conditions will also be avoided. In this research, high-throughput screening will be used to identify improved variants that catalyze desired cyclopropanation reactions at greater rates and with increased stereoselectivity. By developing an expanded set of cyclopropanation biocatalysts with capabilities to act on a wider range of starting materials, the scope and utility of this novel enzymatic reaction will be increased.