Development of Novel Ophthalmic Antibiotics

摘要 细菌性角膜炎是一种威胁视力的疾病，是世界性失明的重要原因。 考虑到疾病的严重性，立即使用广谱抗生素，如第四代抗生素， 氟喹诺酮类药物是成功治疗的关键。然而，阻力上升加上有限的 工业驱动的药物开发突出了对新型眼科抗微生物治疗剂的需求。 该提案的中心目标是通过开发 新型眼用抗生素。初步数据表明，利福平+多粘菌素 B/甲氧苄啶（PT），一种FDA批准的广谱抗生素组合， 协同活性，同时降低产生抗性的倾向。该提案的一个核心目标是 推进利福平+PT的体内疗效研究，以使该联合治疗朝着 FDA的505（B）2快速通道批准流程。然而，鉴于抗生素的快速发展， 因此，必须同时开发新型药物。在这方面， 通过化学文库筛选具有抗假单胞菌活性的新型一流试剂 铜绿假单胞菌和金黄色葡萄球菌是角膜炎的最常见原因， 初步确定了先导化合物。重要的是，筛选模式旨在识别 具有抗生物膜活性的化合物，这是一种众所周知的难以根除的生长状态， 包括眼部感染在内的人类疾病，并且以前不是高通量的目标 药物发现筛选作为命中到铅优先级的一部分，生物和理化研究将 在进行CRISPR干扰必需基因敲低的菌株集时，将创建CRISPR干扰必需基因敲低菌株集，以便 有助于识别先导化合物的细胞靶点，并且角膜炎的小鼠模型将 建立。这后两个工具系统直接使所提出的研究，但也有 在理解角膜炎发病机制中的必需基因/途径方面具有广泛的实用性。 该研究计划提供药物发现的高级培训，同时定义 利福平+PT的临床前景，确定杀死生物膜相关细胞的新型药物， 开发用于抗菌或致病研究的高产工具。此外，该提案还详细说明了 雷切尔·沃兹尼亚克博士的职业发展计划，医学博士，博士，以促进她的过渡， 独立专门的指导、课程工作和专门研讨会将有助于 扩展实验设计和先进研究技术的知识。她获得了 对这些努力的充分的机构和部门支持已经表明， 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.