Biological Treatment of Bacterial Keratitis

项目摘要/摘要 细菌性角膜炎(BK)是一种角膜感染，如果不治疗，可能会导致失明。金黄色葡萄球菌(Sa)， 肺炎链球菌(Sp)和铜绿假单胞菌(PA)是北美BK的三个主要原因。 BK的症状包括受影响的角膜疼痛、发红、炎症、混浊和视力丧失(1)。 AmebaGone Inc.(AG)与UW-Madison眼科部门合作开发一种新的生物防治方法 治疗BK。AG拥有已颁发的专利，并拥有与使用Dictyostelid cell(DC)有关的专业知识，Dictyostelid cell是一种良性的土壤- 已知的通过吞噬作用摧毁病原体的寄居有机体。已颁发的专利广泛涵盖了以下用途 DC食用生物膜包裹和自由生活的(浮游)细菌(“治疗性阿米巴及其用途”；美国专利商标局批准 专利8,551,471和后续专利8,551,671)。一项覆盖欧盟国家的专利已经提交 该公司拥有来自全球3个收藏的3,000多种DC菌株的许可权，这些收藏占据了不同的天然 栖息地。良性DC在10亿多年前从水生阿米巴分化而来，其中一些是致病的。在第一阶段 工作中，AG确定了36种Dicty菌株，能够有效地杀死眼睛上的沙门氏菌(细菌滴度降低4log10) 温度。在这些菌株中，AG鉴定了4株可繁殖萌发到高水平的DC菌株，并开发了一种凝胶 将最大恢复时间从1小时增加到16小时的配方。此外，AG还测试了这些DC的安全性 与载体对照组相比，没有发现角膜病理或不适的增加。在第二阶段项目中 AG建议：1)将DC的目标从针对Sa的单一作用扩大到BK-PA的其他致病因子，并 SP，2)。制定有效的配方并评估其中的直流稳定性。3)进行体内安全性和基础生物学测试 4)眼部直流电治疗。DC鸡尾酒治疗金黄色葡萄球菌、金黄色葡萄球菌或金黄色葡萄球菌感染小鼠角膜的疗效 活着。这些研究中产生的数据将使我们能够准备研究新药(IND)申请 美国食品和药物管理局(FDA)。

Project Summary/Abstract Bacterial keratitis (BK) is an infection of the cornea that, if left untreated, can cause blindness. Staphylococcus aureus (Sa), Streptococcus pneumoniae (Sp), and Pseudomonas aeruginosa (Pa) are three major causes of BK in North America. Symptoms of BK include pain, redness, inflammation, and opacity of the affected cornea and loss of visual acuity (1). AmebaGone Inc., (AG) partnered with UW-Madison Department of Ophthalmology to develop a novel biocontrol method to treat BK. AG holds issued patents and has expert knowledge related to use of Dictyostelid cells (DC), a benign, soil- dwelling organism known to destroy pathogenic bacteria through the phagocytosis. Issued patents broadly cover the use of DC to eat biofilm-enmeshed and free-living (planktonic) bacteria (“Therapeutic Ameba and Uses Thereof”; USPTO granted patent 8,551,471 and follow up patent US 8,551,671). A patent covering countries of the European Union has been filed and the firm has licensed rights to more than 3,000 DC strains from 3 worldwide collections occupying diverse natural habitats. Benign DC diverged from aquatic amoeba, some of which are pathogenic, over 1 billion years ago. In Phase I work, AG identified 36 Dicty strains capable of efficiently killing (> 4 log10 reduction in bacterial titers) Sa at eye temperature. Of these strains, AG identified 4 DC strains that reproducibly germinated to high levels and developed a gel formulation that increased the maximum recovery time from 1 to 16 hours. Additionally, AG tested safety of these DC strains and found no increase in corneal pathology or discomfort compared to the vehicle control. In the Phase II project AG proposes to: 1) Expand the target of DC from singular action against Sa to additional causative agents of BK- Pa and Sp, 2). Develop effective formulation and assess DC stability therein. 3) Conduct in vivo safety and basic biology testing of DC treatment in the eye, and 4). Quantify efficacy of DC cocktails in treating murine cornea infected with Sa, Pa, or Sp in vivo. The data generated in these studies will position us to prepare an Investigational New Drug (IND) application for the Food and Drug Administration (FDA).