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

1个项目摘要 2中耳炎(OM)是儿童健康的主要负担。它是最常见的儿科疾病， 3给美国儿童开抗菌药处方的头号原因。此外，62%的OM儿童表现出病毒感染 4中耳感染，抗生素对这些感染无效，但仍开了处方。广泛使用 对于如此高的发病率和复发率的疾病，使用系统的抗生素被认为会滋生抗生素 6抗性。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未知抗药性的植物天然产物(PNPs)及其衍生物。最先进的合成材料 8我们首创的生物流水线将使抗菌PNPs的优化和规模化生产成为可能 9具有更强的渗透性和抗菌效果。新的PNP化合物将在当地运送到 10使用久经考验的水凝胶，这是一种在室温下易于涂抹的液体，可以快速凝胶。 11，并在接触温鼓膜(TM)时牢固。凝胶内的化学渗透促进剂可以 12克服TM的不渗透障碍，将PNPs带入中耳。我们已经证明了 13单次使用水凝胶制剂就能提供足够的抗生素进行7天的治疗，根除 14在龙猫OM模型中的感染。 15本申请试图完全消除这种流行的儿童疾病中抗生素的使用，并 16通过使用同时治疗细菌和病毒的独立疗法来减轻OM相关的耐药性 17例感染。它专注于黄连素的生产和修饰，小黄碱是一种来自药用植物的生物碱。 黄连素对包括耐甲氧西林金黄色葡萄球菌在内的革兰氏阳性和阴性细菌均有抗菌作用 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即插即用方面的经验 20生物合成在酵母中重建黄连素生产的生物合成途径。我们将结合不同的 21剪裁酶(如卤素酶)和修饰的前体分子以生产修饰的黄连素 22个常规化学合成难以实现的衍生物。治疗效果和疗效观察 23将使用已建立的龙猫OM模型来测试修改后的黄连素衍生物的生物兼容性。 在这个项目中开发的知识和技术将为克服生物 通过分子设计和治疗急性和复发性OM的新方法，在耳朵中设置25个屏障。黄连素的研究重点 26是基于其已知的抗菌作用，一般方法适用于其他PNPs， 27例可以治疗耳部疾病，而不是OM。

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.