Prenylated-Flavin Enzymes as a new Platform for Drug Synthesis and Discovery

异戊二烯化黄素酶作为药物合成和发现的新平台

基本信息

批准号：
9982049
负责人：
Noah Paul Dunham
金额：
$ 6.49万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9982049
关键词：
Acylation Affect Alcohols Aldehydes Alkenes Amides Amines Amino Acid Substitution Anhydrides Aromatic Compounds Biological Biological Process Biology Catalysis Chemicals Collection Complex Coupling Dependence Detection Dimethyl Sulfoxide Directed Molecular Evolution Disease Drug Compounding Effectiveness Engineering Enzymes Esters Exhibits Family Flavins Future Goals Grant Human Knowledge Libraries Life Methods Molecular Mutagenesis Natural Products Organic solvent product Outcome Pathway interactions Patients Pharmaceutical Preparations Philosophy Procedures Process Property Reaction Reagent Research Ring Compound Route Site Solubility Solvents Specificity Structure System Techniques Variant Walking carboxylate carboxylation catalyst cofactor cost cost effective design drug development drug discovery drug synthesis enantiomer high throughput screening human disease improved mutant non-Native novel potassium bicarbonate scaffold screening side effect small molecule stereochemistry technique development thermophilic organism tool

项目摘要

Project Summary/Abstract The discovery and synthesis of small molecules that possess favorable biological properties have markedly enhanced the quality of human life. Current methods for the synthesis of these drug compounds, however, rely heavily on environmentally-harsh solvents and reactions that frequently offer only limited selectivity for the desired outcome. In recent years, enzymes have been increasingly used in conjunction with synthetic reactions to yield products with unparalleled selectivity and without the necessity of harmful solvents. Engineering new enzyme platforms for novel reactivity will expand the collection of biocatalysts that are beneficial to the drug development process, and is thus a goal worth pursuing. Directed evolution has proved to be an unrivaled technique for the development of enzymes that exhibit exquisite catalytic capabilities toward an array of desired reactions. From P450s to TrpB enzymes, many natural scaffolds have been evolved to catalyze reactions that are both known and new to biology, effectively expanding our synthetic toolbox. The research proposed herein aims to engineer prenylated-flavin (prFMN) enzymes by directed evolution to generate biocatalysts that can pave the way to more cost-effective avenues to drug compounds used for the treatment of human diseases. To this end, previously-characterized prFMN enzymes will be applied to a non- native C–C coupling reaction that will ultimately produce chiral allylic and aromatic alcohols. This chemical moiety is found in numerous drug scaffolds; however, the current synthetic procedures often grant insufficient control over the product stereochemistry. A second objective is to apply prFMN enzymes to aromatic acylation reactions, a similar C–C coupling reaction described above that will result in diverse and complex molecular scaffolds. Crafting new synthetic avenues to complex and diversifiable ring structures is important to the drug discovery process by the philosophy of diversity-oriented synthesis. A third objective is to redirect the native prFMN reactivity toward the aromatic carboxylation of large, ring-bearing compounds. Late-stage carboxylation of complex molecular scaffolds can provide another branchpoint to produce libraries of complex and diverse molecules for bioactivity screening and drug discovery. Directed evolution will be applied throughout each aim to engineer enzyme variants capable of catalyzing the chemical transformations with a high degree of specificity. The biocatalysts developed herein may also serve as a platform for future engineering endeavors involving the prFMN cofactor.

项目概要/摘要具有良好生物学特性的小分子的发现和合成已显着然而，目前合成这些药物化合物的方法依赖于。大量使用对环境恶劣的溶剂和反应，这些溶剂和反应通常只能提供有限的选择性近年来，酶越来越多地与合成反应结合使用。生产具有无与伦比的选择性的产品，且无需使用有害溶剂。具有新反应性的酶平台将扩大对药物有益的生物催化剂的收集发展过程，因此是一个值得追求的目标。定向进化已被证明是一种无与伦比的酶开发技术，对一系列所需反应具有出色的催化能力。天然支架已经进化到能够有效地催化生物学已知和新的反应扩展我们的合成工具箱。本文提出的研究旨在通过定向进化来设计异戊二烯化黄素（prFMN）酶生产生物催化剂，为用于药物化合物的更具成本效益的途径铺平道路为此，先前表征的prFMN酶将被应用于非- 天然 C-C 偶联反应，最终产生手性烯丙醇和芳香醇。在许多药物支架中都发现了该部分，然而，目前的合成程序往往不能满足要求。控制产物立体化学的第二个目标是将 prFMN 酶应用于芳香酰化。反应，类似于上述的 C-C 偶联反应，会产生多样化且复杂的分子为复杂且多样化的环结构打造新的合成途径对于该药物非常重要。以多样性为导向的综合哲学的发现过程的第三个目标是重新定向原生。 prFMN 对大型含环化合物的芳香族羧化的反应性。复杂分子支架的构建可以提供另一个分支点来产生复杂多样的文库用于生物活性筛选和药物发现的分子将应用于每个目标。设计能够高度催化化学转化的酶变体本文开发的生物催化剂也可以作为未来工程努力的平台。涉及 prFMN 辅助因子。