Development of chemoenzymatic platform for facile synthesis of alkyl-pyrophosphate analogs

开发化学酶平台以方便合成烷基焦磷酸类似物

• 批准号: 10182076
• 负责人: Shanteri Singh
• 金额: $ 21.1万
• 依托单位: UNIVERSITY OF OKLAHOMA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10182076
• 关键词: Centers of Research Excellence; Clinical; Complex; Coupled; Coupling; Daptomycin; Development; Dimethylallyltranstransferase; Diphosphates; Engineering; Enzymes; Exposure to; FDA approved; Goals; In Situ; Libraries; Methods; Modification; Natural Products; Oklahoma; Organic Synthesis; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Phosphotransferases; Reagent; Structure; Structure-Activity Relationship; Therapeutic; alkyl group; analog; assay development; base; bioactive natural products; chemical synthesis; design; high throughput screening; interest; novel; scaffold; structural biology

Chemoenzymatic natural product diversification is one of the important structural diversification strategies in use to generate pharmaceutically relevant molecules; especially when the molecule to be diversified is structurally complex to be handled by classical chemical synthesis. Our goal is to develop a prenyltransferase (PT)-based chemoenzymatic platform for the late stage structural modification of complex natural product scaffolds and generate compounds with potential therapeutic interest. PTs transfer alkyl groups, which are derived from their activated donors, alkyl pyrophosphates (alkyl-PPs). The current state-of-the-art of synthesis of alkyl-PP analogs relies upon multi-step synthesis and tedious purification methods to separate starting material and other byproducts from the desired product, thereby limiting their practical development as synthetic reagents. Therefore, this proposal seeks to (i) develop a general chemoenzymatic platform for the synthesis and in-situ utilization of alkyl-PP analogs by enzyme engineering approach of two different classes of kinases, and (ii) validate the platform via generating alkyl-diversified library of a FDA-approved macrocyclic drug, daptomycin. The proposed studies will integrate structure determination, high throughput assay development, and structure-guided design to expose structure-activity relationships (SAR) of the selected enzymes. This study will result in highly efficient, robust two-enzymes coupled platform engineered to be efficient in generating novel alkyl-PP donors. Coupling these alkyl-PP analogs with diverse PTs will offer unprecedented access to uniquely bioactive natural product libraries, which are not readily accessible via conventional organic synthesis. Importantly, this study contributes to the discovery of new macrocyclic peptide-based drug leads for downstream pharmaceutical assessment, and clinical use.

化学酶天然产物多样化是用于生成药学相关分子的重要结构多样化策略之一；特别是当要多样化的分子结构复杂而无法通过经典化学合成处理时。我们的目标是开发基于异戊烯基转移酶（PT）的化学酶平台，用于复杂天然产物支架的后期结构修饰，并生成具有潜在治疗意义的化合物。 PT 转移烷基，这些烷基源自其活化的供体，烷基焦磷酸盐 (烷基-PP)。目前最先进的烷基-PP类似物合成技术依赖于多步合成和繁琐的纯化方法来从所需产物中分离起始材料和其他副产物，从而限制了它们作为合成试剂的实际发展。因此，该提案旨在（i）开发一个通用的化学酶平台，通过两种不同类别激酶的酶工程方法合成和原位利用烷基-PP类似物，以及（ii）通过生成 FDA 批准的大环药物达托霉素的烷基多样化库来验证该平台。拟议的研究将整合结构测定、高通量测定开发和结构引导设计，以揭示所选酶的结构-活性关系（SAR）。这项研究将产生高效、稳健的双酶偶联平台，该平台可有效生成新型烷基-PP 供体。将这些烷基-PP类似物与不同的PT偶联将为获得独特的生物活性天然产物库提供前所未有的途径，而这些库是通过传统的有机合成无法轻易获得的。重要的是，这项研究有助于发现新的基于大环肽的药物先导物，用于下游药物评估和临床使用。