Chemoenzymatic synthesis and pharmacological evaluation of designer plant meroterpenoids

设计植物类萜的化学酶合成及药理评价

• 批准号: 10679446
• 负责人: Anna Claire Love
• 金额: $ 6.91万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679446
• 关键词: Address; Anabolism; Angiosperms; Bacteria; Berberine; Biochemical; Biological; Biological Process; Brain; Cannabidiol; Cannabinoids; Cannabis sativa plant; Cells; Chemistry; Communities; Cyclization; Data; Development; Drug Kinetics; Engineering; Enzymes; Escherichia coli; Evaluation; Evolution; Fermentation; Flowers; G-Protein-Coupled Receptors; Generations; Human; Investigation; Learning; Libraries; Lipids; Literature; Metabolic; Minor; Modification; Natural Products; Neurodegenerative Disorders; Obesity; Pathway interactions; Pharmacology; Pharmacology Study; Physiological; Plants; Process; Production; Property; Research Proposals; Route; Scientist; Signal Pathway; Signal Transduction; Source; Streptomyces; Structure; Structure-Activity Relationship; Technology; Terpenes; Testing; Tetrahydrocannabinol; Therapeutic; Yeasts; analog; blood-brain barrier crossing; chemical synthesis; chronic pain management; enzyme pathway; experimental study; fungus; liverwort; manufacturing facility; marine; marine natural product; mutant; non-Native; novel; novel therapeutics; pharmacologic; pharmacophore; phytocannabinoid; polyketide synthase; polyketides; preference; prenyl; prenylation; quinone methide; scaffold; screening; stereochemistry; targeted treatment; therapeutic development

Project Summary/Abstract Land plants like flowering plants and liverworts produce an array of natural products with various biological functions. Polyketide meroterpenoids, comprising structures partially derived from terpenoid biosynthetic pathways and partially derived from polyketide synthase biosynthetic pathways, have attracted scientists for decades from their unique and diverse biological activity. Neuroactive plant meroterpenoids, like phytocannabinoids from Cannabis sativa, represent a particularly exciting suite of compounds with therapeutic promise due to their ability to cross the blood-brain barrier and engage GPCR targets. However, much of the pharmacological data present in the literature to-date has focused on the cannabinoids, Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD), while minor constituents and unique analogs from other producers remain less well studied. Lack of pharmacological data for these compounds is partly due to low accumulation of more rare plant meroterpenoids in native producers, production in less widespread plants (i.e. specific liverwort species), and lack of convergent synthetic routes capable of producing several analogs from one common intermediate. Heterologous production of plant meroterpenoids has been accomplished in eukaryotic hosts (i.e. yeast) but suffers from a pathway bottleneck caused by low catalytic activity and poor expression of plant prenyl cyclization enzymes (e.g. THCA synthase). Bacterially derived cyclization enzymes that generate the same key intermediate, an ortho-quinone methide, provide an attractive alternative for biocatalyst generation toward production of plant-like meroterpenoids and their analogs. Here, I propose the development of new cyclization biocatalysts engineered from bacterial biosynthetic enzymes for chemoenzymatic production of rare and designer plant-like meroterpenoid products and their pharmacological evaluation to assess therapeutic promise. This proposal aims to address issues of supply present for rare meroterpenoids with low accumulation in native producers, generate novel, structurally diverse scaffolds using engineered biocatalysts, and test the pharmacology (i.e. therapeutic promise) of such compounds. In Aim 1, I will engineer biosynthetic pathway enzymes recently identified from the Moore lab, Clz9 and Tcz9, to chemoenzymatically produce meroterpenoids with alternative regioselectivity and steric modification. While neuroactive meroterpenoids are predominantly produced by flowering land plants, other species, like liverworts or marine bacteria, produce similar-looking natural products. In Aim 2, I will identify new prenyl cyclase enzymes from marine bacteria and liverwort sources, expanding the toolkit of biocatalysts for producing plant-like meroterpenoids, especially compounds with unique stereochemistry and larger steric modifications. In Aim 3, produced compounds will be subjected to pre- pharmacokinetics experiments to determine likely metabolic products, and both compounds and their metabolic products will be assessed for ability to activate GPCR targets from the human brain. This research proposal will produce rare and designer plant meroterpenoids using biocatalysts from bacterial hosts and identify which of the produced analogs possess noteworthy therapeutic potential.

项目摘要/摘要 开花植物和利弗沃特（Liverworts）等陆地植物生产各种天然产品 生物功能。聚酮化合物杂货，完成部分衍生自萜类化合物的结构 生物合成途径和部分衍生自聚酮化合物合酶生物合成途径的途径已吸引 科学家数十年来，他们的独特和多样化的生物学活动。神经活性植物的杂化型，例如 来自大麻壁画的植物大麻素代表了一套特别令人兴奋的疗法化合物 由于他们能够越过血脑屏障并参与GPCR目标的能力而承诺。但是，大部分 文献中存在的药理学数据待命于大麻素，Δ9-四氢大麻酚 （Δ9-THC）和大麻二酚（CBD），而其他生产者的次要构成和独特的类似物仍然较少 研究良好。这些化合物缺乏药物数据的部分原因是较少稀有的累积 本地生产者中的植物商制，生产较少的植物（即特定的肝脏物种）， 并且缺乏能够从一个普通中间体产生几个类似物的收敛合成途径。 在真核宿主（即酵母）中，已经完成了植物商制的异源生产 遭受低催化活性引起的途径瓶颈，植物前环化表达不佳 酶（例如THCA合酶）。生成相同钥匙的细菌衍生的环化酶 中级是一种Quinone甲基，为生物催化剂生成提供了一种有吸引力的替代方案 产生类似植物的杂色的元素及其类似物。在这里，我建议开发新的环定 由细菌生物合成酶设计的生物催化剂，用于稀有和稀有的化学酶产生 设计师类植物样的Moroter-Moroteroid产品及其药物评估，以评估治疗承诺。 该提案旨在解决针对稀有机制的供应问题，累积较低 本地生产者，使用工程生物催化剂生成新颖的，结构上不同的脚手架，并测试 这种化合物的药理学（即治疗承诺）。在AIM 1中，我将设计生物合成途径 最近从摩尔实验室CLZ9和TCZ9鉴定的酶以化学酶形式产生杂色 具有替代性调节选择性和空间修改。而神经活性的正规型主要是 由开花的陆地植物，其他物种（如肝脏或海洋细菌）产生的产生相似的外观 天然产品。在AIM 2中，我将确定来自海洋细菌和利弗沃特源的新的蛋白酶环化酶， 扩展生物催化剂的工具包，用于生产类似植物的Moroterpenoids，尤其是具有独特的化合物 立体化学和较大的空间修改。在AIM 3中，生产的化合物将受到预先的影响 药代动力学实验以确定可能的代谢产物，并且化合物及其代谢 将评估产品的能力，以激活人脑的GPCR靶标。这项研究建议将 使用细菌宿主的生物催化剂生产稀有和设计师植物的植物杂化剂，并确定哪个 产生的类似物具有值得注意的治疗潜力。