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产生药物相关分子的重要结构多样化策略之一。尤其是当要多样化的分子在结构上是复杂的，可以通过经典的化学合成来处理。我们的目标是开发基于前阶段的自然产品支架的晚期结构修饰，并产生具有潜在治疗兴趣的化合物，以开发基于前阶段的化学酶平台。 PTS转移烷基，这些烷基源自其活化供体的烷基焦磷酸盐（烷基-PPS）。烷基-PP类似物合成的当前最新作品依赖于多步合成和乏味的纯化方法，将起始材料和其他副产品与所需产品分开，从而将其实际发育限制为合成试剂。因此，该建议试图（i）通过两种不同类别的激酶的酶工程方法开发一个通用的化学酶平台，用于通过酶工程方法合成和原位利用烷基-pp类似物，（ii）通过生成AFDA批准的烷基化的AFDA含量的Macrocrocyclic药物的烷基化库来验证该平台。拟议的研究将整合结构确定，高通量测定开发和结构引导的设计，以揭示所选酶的结构活性关系（SAR）。这项研究将导致高效，可靠的两酶耦合平台，该平台设计为有效地生成新型烷基PP供体。将这些烷基-PP类似物与不同的PT耦合将提供前所未有的访问独特的生物活性天然产物库，这些库无法通过常规的有机合成易于访问。重要的是，这项研究有助于发现新的大环肽铅用于下游药物评估和临床用途。