Protein Mediated Protection from Microbial Keratitis

描述(申请人提供)：微生物性角膜炎是一种严重的眼部感染，可通过角膜疤痕或穿孔导致永久性视力丧失。铜绿假单胞菌是最常见的眼部分离菌之一，已被证明与75%的隐形眼镜介导的感染性角膜炎有关。隐形眼镜配戴者更容易发生微生物角膜炎，开发新的治疗方法是一个重要的研究领域。目前的做法是用抗生素治疗感染，但最近出现的耐药细菌菌株增加了替代治疗的需要。虽然导致微生物角膜炎的确切事件序列尚未完全阐明，但宿主和微生物蛋白水解酶在角膜炎中的作用已有很好的文献记载。本申请建议使用一种新的“保护蛋白”介导的技术作为对抗角膜炎的替代方案。“保护蛋白”是一种新的分子，具有两种不同的活性。第一个明显的活性是通过折叠来稳定蛋白质，第二个不太明显但同样令人印象深刻的保护性蛋白，属于蛋白酶抑制剂类。 我们的假设是，ECI以“保护性蛋白”为特征，αA-晶体蛋白、伽马D-晶体蛋白及其嵌合蛋白胃抑素/亮蛋白/αA-晶体蛋白将抑制宿主和病原体分泌的蛋白水解酶，从而成为对抗微生物角膜炎治疗的良好治疗剂。为SBIR赠款第一阶段设计的具体目标就是基于这一前提，并将为我们提供重要的体外生化分析，从而引导我们进行更有说服力的体内动物模型研究。 我们预计，将这些“保护蛋白”附着在长时间佩戴的软性隐形眼镜上将显著降低与隐形眼镜佩戴者相关的微生物角膜炎的发生率，并可能吸引相当大的市场份额。综上所述，本申请建议使用一种新的“保护蛋白”介导的技术作为对抗角膜炎的替代方案。

DESCRIPTION (provided by applicant): Microbial keratitis is a serious eye infection that can lead to permanent vision loss through corneal scarring or perforation. Pseudomonas aeruginosa is one of the most frequent ocular isolates and has been shown to be associated with 75% of contact lens mediated infectious keratitis. Contact lens wearers are more prone to microbial keratitis and the development of new therapies is an important field of research. The current practice is to treat the infection with antibiotics, but the recent emergence of resistant bacterial strains has heightened the need for alternative treatments. Although the exact sequence of events leading to microbial keratitis has not been fully elucidated, the role of the host and microbial proteases in keratitis has been well documented. This application proposes the use of a novel "Protector protein" mediated technology as an alternative regimen for combating keratitis. "Protector proteins" are novel molecules that have two distinct classes of activities. The first obvious activity is protein stabilization via refolding while the second less obvious but equally impressive group of Protector proteins, belong to the class of protease inhibitors. Our hypothesis is that ECI characterized "Protector proteins"; alphaA-Crystallin, gammaD-Crystallin and the chimeric protein, pepstatin/leupeptin/alphaA-Crystallin will inhibit the proteolytic enzymes secreted by the host and the pathogen, thus serving as a good therapeutic agent against the management of microbial keratitis. The specific aims designed for phase I of SBIR grant, are based on this premise and will provide us with significant in vitro biochemical analysis leading way to more telling in vivo animal model studies. We envisage that attachment of these "Protector proteins" onto extended wear soft contact lenses would significantly reduce the incidence of microbial keratitis associated with contact lens wearers and could attract significant portion of this market. In summary, this application proposes the use of a novel "Protector protein" mediated technology as an alternative regimen for combating keratitis.