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，大自然一直是传统医学健康疗法的源泉，利用植物和真菌来发挥其功效。 2000 多年来，我们现在可以将这些传统疗法的益处归功于自然。 此外，天然产物一直是其生物合成途径中产生的产物。 开发替代疗法的灵感和资源然而，研究表明存在一个问题。 对这些替代品的认识差距根源于缺乏关于其功效的循证信息， 阻碍它们在主流医学中的应用因此，必须制定可以的策略。 允许接触到赋予这些来源药用功效的有价值的分子。 其结构中含有托酚酮环，并已显示出具有治疗癌症的巨大治疗潜力， 疟疾、细菌和真菌感染以及心血管、肾脏和炎症疾病。 托酚酮部分通常充当药效基团，使其成为合成获取和合成的有价值的靶标 然而，芳香七元托酚酮环的复杂性质限制了可持续性。 它们的大规模生产，这反映在常见的合成方法受到低产量、多样化的阻碍 官能团耐受性，以及对危险且昂贵的离线试剂的需求，自然已经进化。 生物催化剂能够直接生成多种功能化的托酚酮，例如真菌 α-酮戊二酸 依赖的非血红素铁双加氧酶 XenC，绕过了传统合成方法的一般挫折。 然而，这种酶促方法的适用性仍然受到底物浓度的阻碍。 XenC 可以容忍所需的两步反应顺序以及有限的底物范围，从而防止 在人们的努力下，这种方法的实际应用获得了大量的托酚酮天然产物。 提高这种生物催化方法的可扩展性、可持续性和更广泛的适用性 生物活性托酚酮，我的目标是将 XenC 改造为一种酶，可以耐受更高的底物负载，在 pH 8 时可持续的一锅级联，并且具有更广泛的非天然底物范围。 酶和开发的生物催化平台将使托酚酮的新型化学酶合成成为可能 倍半萜烯、真菌海洋托酚酮和托酚酮生物碱，提供了可扩展且可持续的策略 从而可以接触到不同的托酚酮和类似物，从而促进其深入的药理学研究。 这将推动新型药物替代品的开发，以治疗人类健康 关注的问题包括感染、炎症、疟疾和癌症。