Development of Novel Ophthalmic Antibiotics

摘要 细菌性角膜炎是一种威胁视力的疾病，是导致世界失明的重要原因。 鉴于疾病的严重性，立即使用广谱抗生素，如第四代 氟喹诺酮类药物是成功治疗的关键。然而，上升的阻力加上有限的 工业驱动的药物开发突出了对新型眼科抗菌疗法的需求。 这项提案的中心目标是通过开发 新型眼科抗生素。初步数据显示利福平多粘菌素前景看好 B/甲氧苄啶(PT)，FDA批准的抗生素组合，显示出广谱， 协同作用，减少了产生抗药性的倾向。这项提议的一个核心目标是 推进利福平PT体内疗效研究，以推动这种组合走向 FDA的505(B)2快速审批流程。然而，鉴于抗生素的使用速度不断加快 除了抗药性，还必须同时开发新的药物类别。在这方面，一个高 筛选具有抗假单胞菌活性的新型一流制剂的生产量化学文库 铜绿假单胞菌和金黄色葡萄球菌是角膜炎的最常见原因，已开始并 初步确定了先导化合物。重要的是，筛查范例旨在确定 具有抗生物被膜活性的化合物，这种生长状态是出了名的难以根除 人类疾病，包括眼部感染，以前并不是高吞吐量的目标 药物发现屏幕。作为Hit-to-Lead优先顺序的一部分，生物和物理化学研究将 将创建CRISPR干扰必需基因敲除菌株集，以便 有助于识别铅化合物的细胞靶标，而角膜炎的小鼠模型将 被建立起来。这后两个工具系统直接支持拟议的研究，但也有 在理解角膜炎发病机制中的必要基因/途径方面具有广泛的实用价值。 该研究计划提供药物发现方面的高级培训，同时将确定 利福平PT的临床前景，识别杀死生物膜相关细胞和 开发用于抗菌药物或致病机理研究的高产量工具。此外，这项提案还详细说明了 雷切尔·沃兹尼亚克博士的职业发展计划，以促进她过渡到 独立。专心致志的指导、课程工作和专业研讨会将有助于 扩大实验设计和先进研究技术的知识。她已经获得了 机构和部门对这些努力的充分支持证明了必要的 成功所必需的技能、职业道德和求知欲。

ABSTRACT Bacterial keratitis is a vision threatening disease and contributes significantly to world blindness. Given the severity of the disease, immediate, broad spectrum antibiotics such as fourth generation fluoroquinolones are crucial for successful treatment. However, rising resistance coupled with limited industry-driven drug development highlights the need for novel ophthalmic antimicrobial therapeutics. The central goals of this proposal are to address the antimicrobial void through the development of novel ophthalmic antibiotics. Preliminary data has demonstrated the promise of rifampicin+polymyxin B/trimethoprim (PT), a combination of FDA-approved antibiotics that displays broad-spectrum, synergistic activity with reduced propensity to develop resistance. A central goal of this proposal is to advance rifampicin+PT with in vivo efficacy studies in order to move this combination toward the FDA’s 505 (B)2 fast track approval process. However given the quickening pace of antibiotic resistance, novel classes of drugs must also be concurrently developed. In that regard, a high throughput chemical library screen for novel, first-in-class agents with activity against Pseudomonas aeruginosa and Staphylococcus aureus, the most common causes of keratitis, has been initiated and initial lead compounds identified. Importantly, the screening paradigm is designed to identify compounds with anti-biofilm activity, a growth state that is notoriously challenging to eradicate in human disease including ocular infections, and has not previously been the target of high throughput drug discovery screens. As part of hit-to-lead prioritization, biologic and physiochemical studies will be performed, a CRISPR-interference essential gene knock down strain set will be created in order to facilitate the identification of cellular targets of lead compounds, and a mouse model of keratitis will be established. These latter two tool systems directly enable the proposed research but also have wide-ranging utility with respect to understanding essential genes/pathways in keratitis pathogenesis. This research program provides advanced training in drug discovery that will simultaneously define the clinical promise of rifampicin+PT, identify novel agents that kill biofilm-associated cells and develop high yield tools for antimicrobial or pathogenesis studies. In addition, this proposal details a career development plan for Dr. Rachel Wozniak, MD, PhD in order to facilitate her transition to independence. Dedicated mentorship, course work and specialized seminars will contribute to an expanding knowledge of experimental design and advanced research techniques. She has obtained full institutional and departmental support for these endeavors had has demonstrated the necessary skill set, work ethic and intellectual curiosity necessary for success.