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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要开花植物和地钱等陆地植物可产生一系列天然产物，其中含有多种成分。生物功能聚酮化合物类萜，包含部分源自萜类化合物的结构。生物合成途径，部分源自聚酮合酶生物合成途径，吸引了几十年来，科学家们从其独特而多样的生物活性中发现了神经活性植物类萜，例如。来自大麻的植物大麻素代表了一组特别令人兴奋的具有治疗作用的化合物由于它们能够穿过血脑屏障并与 GPCR 靶点结合，因此前景广阔。迄今为止文献中的药理学数据主要集中在大麻素，Δ9-四氢大麻酚 (Δ9-THC) 和大麻二酚 (CBD)，而其他生产商的次要成分和独特类似物仍然较少缺乏对这些化合物的充分研究的药理学数据部分是由于更稀有的化合物积累较少。本地生产者中的植物类萜，在不太广泛分布的植物中生产（即特定的苔类植物），缺乏能够从一种常见中间体生产多种类似物的聚合合成路线。植物类萜的异源生产已在真核宿主（即酵母）中完成，但植物异戊二烯环化催化活性低、表达差导致途径瓶颈产生相同密钥的细菌来源的环化酶。中间体，一种邻醌甲基化物，为生物催化剂的生成提供了一种有吸引力的替代品在这里，我建议开发新的环化方法。由细菌生物合成酶设计的生物催化剂，用于化学酶法生产稀有和设计类植物类萜产品及其药理学评估，以评估治疗前景。该提案旨在解决稀有类萜的供应问题，且其积累量较低。本地生产者，使用工程生物催化剂生成新颖的、结构多样的支架，并测试在目标 1 中，我将设计生物合成途径。 Moore 实验室最近鉴定出 Clz9 和 Tcz9 酶，可通过化学酶法产生类萜具有替代区域选择性和空间修饰，而神经活性类萜主要是。由开花的陆地植物产生，其他物种，如地钱或海洋细菌，也产生类似的外观在目标 2 中，我将从海洋细菌和地钱中鉴定出新的异戊二烯环化酶，扩大生物催化剂的工具包，用于生产类植物类萜，特别是具有独特特性的化合物在目标 3 中，所产生的化合物将进行预-立体化学和更大的空间修饰。药代动力学实验以确定可能的代谢产物以及化合物及其代谢该研究计划将评估产品激活人脑 GPCR 靶标的能力。使用来自细菌宿主的生物催化剂生产稀有和设计的植物类萜，并确定哪些产生的类似物具有显着的治疗潜力。