Live-Cells Assays for Lysosomal Enzyme Activity

描述（由申请人提供）：该小型企业创新研究I期项目旨在开发能够测量活细胞和组织中溶酶体酶活性的新型靶向荧光底物。如果成功，拟议的研究将提供所需的突破，以推动发现有前途的新疗法和调节药物，用于溶酶体贮积病和相关的医学应用。在该项目的第一阶段，Marker Gene Technologies，Inc.将通过制备用于溶酶体酶的新的荧光糖苷酶底物并证明来自正常或疾病来源的活细胞中的差异染色来确定该技术的可行性。在II期，将在体外和体内测定这些新底物对活细胞中溶酶体酶的特异性和局部抑制或诱导以及以细胞或组织特异性方式区分单个酶活性的能力。新的底物和由此产生的检测系统将提供创新的方法来定量溶酶体酶的功能，并筛选二级药物或蛋白质给药的影响，使其成为各种重要的医学应用的有用的分析工具。该项目的成功为溶酶体贮积病的医学干预领域开辟了巨大的商业可能性，如Sandhoff病，Tay-Sachs综合征，Krabb？的疾病和戈谢氏病，以及在细胞培养系统中筛选新的蛋白质和药物用于调节这些疾病中的溶酶体酶活性的功效，以及开发新的，通用和特定的高通量溶酶体酶检测策略。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.