Sustainable biocatalytic generation of bioactive tropolone natural products

生物活性托酚酮天然产物的可持续生物催化生成

• 批准号: 10679828
• 负责人: Jose R. Hernandez-Melendez
• 金额: $ 4.03万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679828
• 关键词: Address; Alkaloids; Alternative Medicine; Anthelmintics; Anti-Bacterial Agents; Antifungal Agents; Antimalarials; Antimitotic Agents; Bacterial Infections; Biological; Biomimetics; Breathing; Bypass; Cardiovascular Diseases; Chemicals; Colchicine; Complex; Development; Dioxygenases; Directed Molecular Evolution; Disease; Engineering; Enzymes; Generations; Goals; Health; Human; Infection; Inflammation; Inflammatory; Iron; Kidney Diseases; Knowledge; Libraries; Light; Mainstreaming; Malaria; Malignant Neoplasms; Medicine; Metals; Methods; Mycoses; Natural Products; Nature; Pathway interactions; Pattern; Pharmacology Study; Plants; Property; Reaction; Reagent; Research; Resources; Route; Sesquiterpenes; Source; Structure; Therapeutic; Traditional Medicine; Tropolone; Variant; Walking; Work; alpha ketoglutarate; analog; anti-cancer; bioactive natural products; catalyst; chelation; cost; cycloaddition; drug candidate; drug discovery; evidence base; functional group; fungus; human disease; improved; interest; large scale production; malaria infection; marine; member; metalloenzyme; nanomolar; non-Native; novel; pharmacophore; physical property; practical application; prevent; scaffold; success

PROPOSAL SUMMARY For years, nature has been the source for health remedies in traditional medicine, using plants and fungi for their curative benefits for over 2000 years. We can now attribute the benefits of these traditional treatments to natural products generated in their biosynthetic pathways. Moreover, natural products have been a consistent source of inspiration and resource in the development of alternative therapeutics. However, studies show that there is a gap in knowledge on these alternatives which is rooted in a lack of evidence-based information on their efficacy, hindering their application in mainstream medicine. Therefore, it is imperative to develop strategies that could allow to reach the valuable molecules that give these sources their medicinal power. A subset of natural products contains tropolone rings in their structures and have shown to have great therapeutic potential for treating cancer, malaria, bacterial and fungal infections as well as cardiovascular, renal, and inflammatory diseases. The tropolone moiety commonly acts as a pharmacophore, making it a valuable target to synthetically access and evaluate. However, the complex nature of the aromatic seven-membered tropolone ring limits the sustainability of their large-scale production, this reflected in common synthetic methods being hindered by low yields, diverse functional group tolerance, and the need for hazardous and costly reagents. Conversely, nature has evolved biocatalysts that enable direct routes to diversely functionalized tropolones such as the fungal α-ketoglutarate dependent non-heme iron dioxygenase XenC, bypassing the general setbacks of traditional synthetic methods. Nonetheless, the applicability of this enzymatic method remains hindered by the concentration of substrate that XenC can tolerate, the required two-step reaction sequence, and the limited substrate scope that prevents the practical applications of this approach to access a plethora of tropolone natural products. In the efforts of improving the scalability, sustainability, and broader applicability of this biocatalytic method towards accessing bioactive tropolones, I aim to engineer XenC into an enzyme that tolerates higher substrate loadings, works in a sustainable one-pot cascaded at pH 8, and has a broader non-native substrate scope. The successfully evolved enzymes and the developed biocatalytic platform will enable novel chemoenzymatic syntheses of tropolone sesquiterpenes, fungal marine tropolones, and tropolone alkaloids, providing a scalable and sustainable strategy that allows to reach diverse tropolones and analogs to facilitate their in-depth pharmacological studies. In turn, this will impulse the development of novel medicine alternatives with the purpose of treating human-health concerns including infections, inflammation, malaria, and cancer.

提案摘要 多年来，自然一直是传统医学中健康疗法的来源，使用植物和真菌为其 超过2000年的治疗益处。现在，我们可以将这些传统疗法的好处归因于自然 在其生物合成途径中产生的产品。此外，天然产品一直是一致的来源 替代疗法发展的灵感和资源。但是，研究表明有一个 知识差距是这些替代方案的差距，这根源于缺乏基于证据的信息， 阻碍他们在主流医学中的应用。因此，必须制定可以 允许达到赋予这些来源医疗能力的宝贵分子。天然产品的子集 在其结构中包含Tropolone环，并且已证明具有巨大的治疗癌症潜力， 疟疾，细菌和真菌感染以及心血管，肾脏和炎症性疾病。 Tropolone部分通常充当药理，使其成为合成访问和的宝贵目标 评价。但是，芳香七元的Tropolone环的复杂性限制了可持续性 在它们的大规模生产中，这种反映在常见的合成方法中受到低收益的阻碍，潜水员 功能组的耐受性以及对危险和昂贵的试剂的需求。相反，大自然发展了 生物催化剂，可以直接路由到多样化的型号，例如真菌α-酮戊二酸酯 依赖性非血红素双加氧酶Xenc，绕过传统合成方法的一般挫折。 尽管如此，这种酶方法的适用性仍然受到底物浓度的阻碍 XENC可以忍受，所需的两步反应序列，以及有限的基材范围，以防止 这种方法可用于访问众多热带天然产品的实际应用。努力 提高这种生物催化方法访问的可伸缩性，可持续性和更广泛的适用性 我的目标是生物活性的tropolones，将Xenc设计成一种耐受较高底物载荷的酶，可在 可持续的一锅在pH 8处级联，并具有更广泛的非本地底物范围。成功发展 酶和开发的生物催化平台将启用Tropolone的新型化学酶合成 倍苯二酚，真菌海洋Tropolones和Tropolone生物碱，提供可扩展且可持续的策略 这允许到达潜水员的轨道和类似物，以促进其深入的药物研究。反过来， 这将冲动开发新型医学替代品，目的是治疗人类健康 包括感染，感染，疟疾和癌症的问题。