Chemoenzymatic synthesis of pradimicin analogues for novel antifungal and antivir

描述（申请人提供）：由病原微生物、病毒或寄生虫引起的传染病是对人类健康的主要威胁，每年导致全球16.2%的死亡。迫切需要发现有效的治疗方法来对抗这些疾病并挽救生命。Pradimicin A是来自Actinomadura hibisca P157-2的抗真菌和抗病毒天然产物。普拉米星的水溶性差和副作用阻碍了其发展成为抗感染药物。为了利用普拉米星A的重要生物活性并找到有前途的抗感染候选药物，该学术研究增强奖（AREA）R15应用程序是研究普拉米星A的生物合成途径，并使用酶和化学方法生成新的普拉米星类似物。提出的研究的三个中心假设是：1）完整的普拉米星A生物合成途径可以在异源宿主中重建和工程化; 2）外源糖部分和相应的糖基转移酶可以工程化到普拉米星生物合成途径中;和3）新的普拉米星类似物，例如具有不同氨基糖的那些具有抗真菌和抗病毒活性。研究人员将测试这些中心假设，并通过追求四个具体目标来实现本项目的总体目标：1）研究普拉米星A生物合成中的后期剪裁步骤; 2）设计新型普拉米星类似物的生物合成; 3）化学合成新型普拉米星糖苷; 4）评价合成的普拉米星类似物的抗真菌和抗病毒活性。生物合成工程与化学合成相结合，将产生一系列“非天然”的普拉米星类似物，可能为新的抗感染药物发现提供新的先导化合物。主要研究者和两名合作研究者分别来自生物工程、化学和生物化学以及生物学三个不同的部门。两名研究生和四名本科生将在技术人员的协助下进行拟议的研究。这个多部门的合作项目将为学生研究人员提供出色的培训机会，按照R15补助金目标。学生将受益于这个跨学科的项目，与来自不同部门和不同背景的科学家合作。预计该项目将促进犹他州州立大学在生物医学研究方面的合作，参与的学生将获得抗感染药物发现的实践经验。 公共卫生相关性：该项目旨在使用酶和化学方法的组合来产生新型普拉米星类似物的库，并评估这些合成化合物的抗真菌和抗病毒活性。该提案的目标与NIH学术研究增强奖（AREA）R15计划的目标一致，并将通过为新的抗感染药物发现提供有前途的分子对公共卫生产生重大影响。这个多部门的项目将由一个研究团队进行，该团队由三名教师，一名技术人员，两名研究生和四名本科生组成。

DESCRIPTION (provided by applicant): Infectious diseases caused by pathogenic microorganisms, viruses or parasites are a major threat to human health, and lead to 16.2% of the world's annual deaths. Discovery of effective therapeutics is urgently needed to fight these diseases and save lives. Pradimicin A is an antifungal and antiviral natural product from Actinomadura hibisca P157-2. Development of pradimicins into anti-infective drugs has been hampered by their poor water solubility and side effects. To take advantage of the significant biological activities of pradimicin A and find promising anti-infective drug candidates, this Academic Research Enhancement Award (AREA) R15 application is to study the biosynthetic pathway of pradimicin A and generate novel pradimicin analogs using both enzymatic and chemical methods. Three central hypotheses for the proposed research are: 1) The complete pradimicin A biosynthetic pathway can be reconstituted and engineered in a heterologous host; 2) Foreign sugar moieties and the corresponding glycosyltransferases can be engineered into the pradimicin biosynthetic pathway; and 3) New pradimicin analogs such as those with different amino sugars have antifungal and antiviral activities. The investigators will test these central hypotheses and accomplish the overall objective of this project by pursuing four specific aims: 1) Investigate the late tailoring steps in the biosynthesis of pradimicin A; 2) Engineer the biosynthesis of novel pradimicin analogs; 3) Chemically synthesize novel pradimicin glycosides; and 4) Evaluate the synthesized pradimicin analogs for antifungal and antiviral activities. With the combination of biosynthetic engineering and chemical synthesis, a series of "unnatural" pradimicin analogs will be generated, which may provide novel lead compounds for new anti- infective drug discovery. The PI and two Co-investigators are from three different departments including biological engineering, chemistry and biochemistry, and biology, respectively. Two graduate and four undergraduate students will perform the proposed research, with assistance from a technician. This multi-departmental collaborative project will provide outstanding training opportunities for student researchers, in accordance with R15 grant objectives. The students will benefit from this interdisciplinary project by working with scientists from different departments and with various backgrounds. It is expected that this project will promote collaborative efforts in biomedical research at Utah State University and the involved students will gain hands-on experience in anti- infective drug discovery. PUBLIC HEALTH RELEVANCE: This project aims to generate a library of novel pradimicin analogs using a combination of enzymatic and chemical methods, and evaluate the antifungal and antiviral activities of these synthesized compounds. The objectives of this proposal are consistent with the goals of the NIH Academic Research Enhancement Award (AREA) R15 program, and will have a significant impact on public health by providing promising molecules for new anti-infective drug discovery. This multi-departmental project will be conducted by a team of researchers consisting of three faculty members, a technician, two graduate students, and four undergraduate students.