High-Throughput Solid-Phase Combinatorial Biocatalysis

描述(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，为选择性筛选和将在这些筛选中测试的新化合物提供了新的药物靶点。虽然组合方法已经产生了大量的化合物库，但通常这些化合物会产生改进的候选先导化合物，这些候选化合物必须经过传统药物化学的进一步转化才能产生新的候选药物。在高通量组合方法学的背景下，生物工程对铅优化的影响没有它对铅发现的影响那么大，主要是因为优化铅化合物往往需要高选择性、复杂的化学成分，而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致选择性和独特的反应活性来帮助克服这些障碍；然而，到目前为止，这项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，包括与固体和可溶性聚合物载体相连的铅分子的反应和文库的生成。在此过程中，我们将开发一种高通量、生物催化的药物发现技术。具体目标是： 1.扩大固体和聚合物载体化合物在水和非水介质中的生物催化的广度； 2.开发连接先导化合物和将其衍生物从固体和聚合物载体上移除的策略； 3.展示高通量的组合生物催化先导优化天然和合成的复杂分子，筛选生物活性的衍生物，并利用生物转化放大结构和功能上感兴趣的衍生物。这项工作将使用一系列的先导分子，从附着在固体和可溶聚合物载体上的酶底物到复杂的化合物(类黄酮类岩白菜素和目前的HIV-1蛋白酶抑制剂吲地那韦)。这项研究计划的成功完成将带来一种强大的方法学，生物医学研究人员可以使用这种方法学来寻找新的、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.