Simultaneous Treatment of Viral and Bacterial Otitis Media Using Plant Natural Products

1项目概要 2中耳炎（OM）是儿童的主要健康负担。它是最常见的儿科疾病， 3美国儿童使用抗菌药物处方的首要原因。此外，62%的OM儿童表现出病毒感染， 4中耳感染，抗生素无效，但仍然处方。广泛使用 针对这种高患病率和复发率的疾病，全身使用抗生素被认为会滋生抗生素 6抵抗为了同时治疗病毒和细菌感染，我们将设计和合成复杂的 7种植物天然产物（PNP）及其衍生物，无已知耐药性。最先进的合成 我们开创的8条生物管道将使抗菌PNPs的优化和可扩展生产成为可能 9具有增强的渗透性和抗微生物功效。新的PNP化合物将在当地输送到 10中耳使用一种经过验证的水凝胶，这是一种易于在室温下涂抹的液体， 11并且在接触热鼓膜（TM）时牢固。凝胶罐内的化学渗透促进剂 12克服TM的不可渗透屏障并将PNP带入中耳。我们已经证明 13，水凝胶制剂的单次应用提供了足够的抗生素用于7天的治疗，根除了 图14灰鼠OM模型中的感染。 本申请试图完全消除这种流行的儿童疾病中的抗生素使用， 16通过使用治疗细菌和病毒的独立疗法来减轻OM相关的耐药性 17感染它专注于小檗碱的生产和修饰，小檗碱是一种来自药用植物的生物碱。 黄连素对革兰氏阳性菌和阴性菌（包括MRSA）具有抗菌作用 19，并显示出对多种病毒的抗病毒特性。这项工作将利用我们在PNP方面的经验 20生物合成以在酵母中重建用于小檗碱生产的生物合成途径。我们将结合不同的 21种剪裁酶（例如，卤化酶）和修饰的前体分子以产生修饰的小檗碱 22衍生物是难以实现的常规化学合成。的治疗有效性和 将使用已建立的灰鼠OM模型测试修饰的黄连素衍生物的生物相容性。 24.本项目中开发的知识和技术将为克服生物学障碍提供新的见解。 通过分子设计和新的治疗急性和复发性OM的耳朵中的25个障碍。关注黄连素 26是基于其已知的抗微生物作用，并且一般方法适用于 27可以治疗OM以外的耳部疾病。

1 Project Summary 2 Otitis media (OM) is a major child health burden. It is the most commonly diagnosed pediatric disease and the 3 #1 reason for antimicrobial prescription to US children. Moreover, 62% of children with OM demonstrate viral 4 infections in their middle ear, to which antibiotics are ineffective but prescribed nonetheless. The widespread use 5 of systemic antibiotics against a disease of such high prevalence and recurrence is believed to breed antibiotic 6 resistance. To treat viral and bacterial infections simultaneously, we will design and synthesize sophisticated 7 plant natural products (PNPs) and their derivatives with no known drug resistance,. The state-of-the-art synthetic 8 biology pipeline that we pioneered will enable the optimization and scalable production of antimicrobial PNPs 9 with enhanced permeability and antimicrobial efficacy. The novel PNP compounds will be delivered locally into 10 the middle ear using a proven hydrogel, which is an easy-to-apply liquid at room temperature and gels quickly 11 and firmly upon contacting warm tympanic membrane (TM). Chemical permeation enhancers inside the gel can 12 overcome the impermeable barrier of the TM and bring the PNPs into the middle ear. We have demonstrated 13 that a single application of the hydrogel formulation provides enough antibiotics for a 7-day treatment, eradicating 14 the infection in a chinchilla OM model. 15 The current application attempts to completely eliminate antibiotic usage in this prevalent childhood disease and 16 to mitigate OM-related drug resistance by using a stand-alone therapy that treats both bacterial and viral 17 infections. It focuses on the production and modification of berberine, an alkaloid derived from medicinal plants. 18 Berberine has demonstrated antimicrobial effects against Gram-positive and -negative bacteria including MRSA 19 and has shown antiviral properties against multiple viruses. This work will leverage our experience in PNP 20 biosynthesis to reconstruct a biosynthetic pathway in yeast for berberine production. We will incorporate diverse 21 tailoring enzymes (e.g., halogenases) and modified precursor molecules to produce modified berberine 22 derivatives that are difficult to achieve by conventional chemical synthesis. The therapeutic efficacy and 23 biocompatibility of the modified berberine derivatives will be tested using an established chinchilla OM model. 24 The knowledge and techniques developed in this project will provide new insight into overcoming the biological 25 barriers in the ear via molecular design and new treatments for acute and recurrent OM. The focus on berberine 26 is based on its known antimicrobial effect, and the general methodologies are applicable to other PNPs that 27 could treat ear diseases beyond OM.