Biosynthesis of novel and natural plant alkaloid compounds for drug production and discovery

用于药物生产和发现的新型天然植物生物碱化合物的生物合成

• 批准号: 9254663
• 负责人: Kristy Hawkins
• 金额: $ 22.5万
• 依托单位: ANTHEIA, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-15 至 2018-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9254663
• 关键词: Alkaloids; Anabolism; Antitussive Agents; CRISPR/Cas technology; Cellular Metabolic Process; Chemicals; Chromosomes; Climate; Complex; Coughing; Cytochrome P450; DNA cassette; Development; Disease; Endoplasmic Reticulum; Engineering; Ensure; Enzymes; Evaluation; FDA approved; Foundations; Generations; Geography; Industrialization; Legal; Link; Malignant Neoplasms; Medicine; Membrane; Methods; Microbe; Modification; Morphinans; Natural Products; Nature; Neurodegenerative Disorders; Noscapine; Opioid; Opium; Papaver; Parents; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Plant alkaloid; Plants; Politics; Production; Property; Proteins; Publishing; Regulation; Research; Route; Saccharomyces cerevisiae; Science; Secure; Small Business Innovation Research Grant; Source; Structure; Techniques; Technology; Time; Tyrosine; Yeasts; base; cofactor; combinatorial; commercialization; cost; cost effective; drug candidate; drug discovery; drug production; enzyme biosynthesis; enzyme pathway; experimental study; feeding; genome editing; homologous recombination; improved; innovation; manufacturing process; metabolic engineering; microbial; microbial host; milligram; novel; novel strategies; novel therapeutics; promoter; scaffold; scale up; sugar

Abstract Plants produce an exceptional diversity and number of natural products, many of which are unique to the plant kingdom. However, the difficulties in sourcing medicinal compounds from native plant hosts in sufficient quantity and diversity present a critical barrier to supplying existing medicines, identifying new drug candidates, and progressing through the path to FDA- approved pharmaceuticals. Noscapine is an established antitussive API currently under evaluation to treat a number of diseases including cancer and neurodegenerative disorders. Noscapine is only available via extraction from opium poppy and the need to source this compound through a drug crop leads to increased cost and regulation, which could be avoided if an alternative source was developed that was not inherently linked to the production of controlled substances. A novel approach will be developed for sourcing this and related compounds, by engineering noscapine biosynthesis from sugar for the first time in an industrial microbe, Baker's yeast. By utilizing a “clean” host that is highly amenable to metabolic engineering and industrial-scale production, this approach will establish a constant, secure, and abundant supply of noscapine and related medicinal compounds that is free from contaminating controlled substances and unnecessary regulation. The objectives of this Phase I project will be to develop engineered yeast strains that produce high levels of natural and non-natural noscapinoids, secoberberines, and other valuable alkaloids, thereby providing microbial platforms for the biosynthesis of diverse plant- based medicinal compounds. To achieve high-level production of the target alkaloids, the rate- limiting activities of five pathway cytochrome P450 enzymes will be optimized. In addition, substrate-feeding experiments that target the incorporation of both early precursor molecules and later-stage scaffolds, will enable the synthesis of numerous novel, non-natural BIA molecules and provide key information on the substrate promiscuity of the enzymes for later combinatorial engineering strategies. Through the development of a disruptive and safe technology for producing an important class of plant alkaloids, the deliverables from this project will open up access to a diverse array of natural plant alkaloid compounds and an even greater number of non-natural derivatives. The successful completion of this SBIR project will provide proof-of-concept for a new commercial manufacturing process for established plant-based medicines and a combinatorial biosynthesis platform for the generation of novel drug molecules.

摘要 植物产生异常多样性和数量的天然产品，其中许多 是植物界所独有的然而，在采购药用化合物的困难， 足够数量和多样性的本地植物宿主是供应的关键障碍， 现有的药物，确定新的候选药物，并通过FDA的途径取得进展- 批准的药品。诺斯卡品是一种成熟的止咳API，目前正在 用于治疗包括癌症和神经退行性疾病在内的多种疾病的评估。 诺斯卡品只能从罂粟中提取， 通过药物作物生产化合物会导致成本和监管增加，如果 开发了一种与生产无内在联系的替代来源， 管制物质。将开发一种新的方法， 化合物，通过工程诺斯卡品生物合成糖首次在工业 面包酵母通过利用高度适合代谢的“清洁”宿主， 工程和工业规模的生产，这种方法将建立一个恒定的，安全的， 无污染的诺斯卡品和相关药用化合物的充足供应 管制物质和不必要的管制。 第一阶段项目的目标是开发工程酵母菌株， 产生高水平的天然和非天然类诺斯卡品，secoberberines，和其他 有价值的生物碱，从而为生物合成不同的植物提供微生物平台， 药用化合物。为实现目标生物碱的高水平生产， 将优化五种途径细胞色素P450酶的限制活性。此外，本发明还提供了一种方法， 靶向掺入两种早期前体分子的底物进料实验 和后期支架，将能够合成许多新的，非天然的BIA 分子，并提供有关酶的底物混杂性的关键信息， 组合工程策略通过开发颠覆性且安全的 生产一类重要的植物生物碱的技术，该项目的可交付成果 将开辟获得多种天然植物生物碱化合物的途径， 非自然衍生物的数量。该SBIR项目的成功完成将提供 一个新的商业制造工艺的概念验证， 药物和用于产生新型药物分子的组合生物合成平台。