Live-Cells Assays for Lysosomal Enzyme Activity

描述（由申请人提供）：这个小企业创新研究第一阶段项目旨在开发新的靶向荧光底物，能够测量活细胞和组织中的溶酶体酶活性。如果成功，拟议的研究将提供所需的突破，以推动发现有希望的新疗法和溶酶体贮积病的调节药物和相关的医疗应用。在该项目的第一阶段，Marker Gene Technologies, Inc.将通过为溶酶体酶制备新的荧光糖苷酶底物，并证明在正常或疾病来源的活细胞中进行差异染色，来确定该技术的可行性。在第二阶段，这些新的底物将在体外和体内进行测试，以确定它们在活细胞中对溶酶体酶的特异性和局部抑制或诱导能力，以及以细胞或组织特异性的方式区分单个酶的活性。新的底物和由此产生的检测系统将为定量测定溶酶体酶功能和筛选二次药物或蛋白质给药的影响提供创新方法，使其成为各种重要医学应用的有用分析工具。该项目的成功为溶酶体贮积性疾病（如Sandhoff病、Tay-Sachs综合征、Krabb病和戈谢病）的医学干预领域开辟了巨大的商业可能性，也为细胞培养系统中调节这些疾病溶酶体酶活性的新蛋白质和药物的筛选，以及开发新的、通用的和特定的高通量溶酶体酶检测策略开辟了巨大的商业可能性。此外，它将导致这些领域的商业和许可产品。

DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase I project aims to develop new targeted fluorogenic substrates capable of measuring lysosomal enzyme activity in living cells and tissues. If successful, the proposed research will provide breakthroughs needed to advance the discovery of promising new therapies and modulating drugs for lysosomal storage diseases and allied medical applications. In Phase I of this project, Marker Gene Technologies, Inc. will establish the feasibility of the technology by preparing new fluorogenic glycosidase substrates for lysosomal enzymes and demonstrating differential staining in living cells that are from normal or are of disease origin. In Phase II, these new substrates will be assayed in vitro and in vivo for their ability to measure specific and localized inhibition or induction of lysosomal enzymes in living cells as well as differentiate individual enzyme activities in a cell- or tissue-specific manner. The new substrates and the resulting detection systems will provide innovative methods to quantitate lysosomal enzyme function and to screen for the influence of secondary drug or protein administration, making them useful analytical tools for a variety of significant medical applications. The success of this project opens up enormous commercial possibilities in the fields of medical intervention in lysosomal storage diseases such as Sandhoff's disease, Tay-Sachs syndrome, Krabb¿'s disease and Gaucher's disease, as well as in the screening of new proteins and drugs in cell-culture systems for efficacy in modulating lysosomal enzyme activity in these diseases, and development of new, general and specific high-throughput lysosomal enzyme detection strategies. In addition, it will lead to commercial and licensable products in these areas.