High-Throughput Solid-Phase Combinatorial Biocatalysis

描述（由申请人提供）：基因组学、蛋白质组学和组合化学的快速发展重塑了药物发现领域，为选择性筛选提供了新的药物靶点，并为这些筛选提供了待测试的新化合物。虽然组合方法已经产生了大量的化合物库，但通常这些化合物导致改进的先导候选物，其必须通过常规药物化学进行进一步转化以产生新的候选药物。在高通量组合方法的背景下，生物工程对先导化合物优化的影响几乎不如对先导化合物发现的影响大，这主要是因为优化先导化合物通常需要高度选择性、复杂的化学反应，并且缺乏适当广泛的高通量平台。组合生物催化可以通过利用酶和微生物生物催化剂的精细选择性和独特反应性来帮助克服这些障碍;然而，迄今为止，该技术仅限于可溶性底物的衍生化。我们建议扩大组合生物催化的范围，包括反应，并产生图书馆，铅分子连接到固体和可溶性聚合物支持。在这个过程中，我们将开发一种用于药物发现的高通量生物催化技术。具体目标是： 1.在水和非水介质中扩大固体和聚合物载体化合物的生物催化范围; 2.制定从固体和聚合物载体上附着铅化合物和去除其衍生物的战略; 3.为了证明高通量，组合生物催化导致复杂的天然和合成分子的优化，筛选所得衍生物的生物活性，并使用生物转化放大结构和功能上有趣的衍生物。一系列的铅分子将被用于这项工作，从酶底物，附着在固体和可溶性聚合物支持复杂的化合物（类黄酮岩白菜素和目前的HIV-1蛋白酶抑制剂茚地那韦）。该研究计划的成功完成将产生一种强大的方法，可供生物医学研究人员用于寻找新的，更有效的小分子疗法。

DESCRIPTION (provided by applicant): Rapid developments in genomics, proteomics, and combinatorial chemistry have reshaped the field of drug discovery, providing new drug targets for selective screens and new compounds to be tested in those screens. While combinatorial methods have given rise to large libraries of compounds, typically these compounds result in improved lead candidates that must undergo further transformations by conventional medicinal chemistry to yield new drug candidates. Bioengineering, in the context of high-throughput combinatorial methodologies, has not impacted lead optimization nearly as much as it has lead discovery, mainly because of the highly selective, intricate chemistries often required to optimize lead compounds and the lack of a suitably broad high-throughput platform. Combinatorial biocatalysis can help overcome these obstacles by exploiting the exquisite selectivity and unique reactivity of enzymes and microbial biocatalysts; however, to date this technology has been limited to the derivatization of soluble substrates. We propose to expand the scope of combinatorial biocatalysis to include reactions on, and the generation of libraries from, lead molecules attached to solid and soluble polymer supports. In the process, we will develop a high-throughput, biocatalytic technology for drug discovery. The specific aims are: 1. To expand the breadth of biocatalysis on solid- and polymer-supported compounds in aqueous and nonaqueous media; 2. To develop strategies for attaching lead compounds and removing their derivatives from solid and polymeric supports; 3. To demonstrate high-throughput, combinatorial biocatalytic lead optimization of complex natural and synthetic molecules, screen resulting derivatives for biological activity, and scale up structurally and functionally interesting derivatives using biotransformations. A series of lead molecules will be used in this work, ranging from enzyme substrates that are attached onto solid and soluble polymer supports to complex compounds (the flavonoid bergenin and the current HIV-1 protease inhibitor indinavir). Successful completion of this research program will result in a powerful methodology that can be used by biomedical investigators in the search for new, more potent small molecule therapeutics.