Live-Cell Assays for Lysosomal Enzyme Activity

溶酶体酶活性的活细胞测定

• 批准号: 8252668
• 负责人: JOHN J NALEWAY
• 金额: $ 50.63万
• 依托单位: MARKER GENE TECHNOLOGIES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8252668
• 关键词: Acid Phosphatase; Ally; Area; Biochemical; Biological Assay; Biological Monitoring; Biotechnology; Cathepsins; Cell Culture Techniques; Cells; Clinical; Collaborations; Cytolysis; Detection; Diagnostic; Diagnostic tests; Disease; Disease Progression; Drug usage; Effectiveness; Enzymes; Evaluation; Fluorescence; Fluorescence Microscopy; Fluorescence Spectroscopy; Fluorogenic Substrate; Fucosidase; Galactosidase; Gaucher Disease; Genes; Glycoside Hydrolases; Glycosphingolipids; Goals; In Vitro; Individual; Industry; Institution; Intervention; Laboratories; Laboratory Markers; Laboratory Research; Lead; Leucine Aminopeptidase; Libraries; Life; Lipase; Lysosomal Storage Diseases; Lysosomes; Mannosidase; Marketing; Measurement; Measures; Medical; Medical Research; Methods; Monitor; Morphologic artifacts; Neuraminidase; Normal tissue morphology; Organelles; Output; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phosphoric Monoester Hydrolases; Proteins; Protocols documentation; Radiolabeled; Reporting; Reproducibility; Research; Sandhoff Disease; Screening procedure; Small Business Innovation Research Grant; Staining method; Stains; Sulfatases; Syndrome; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Tissues; Treatment Efficacy; Treatment Protocols; United States National Institutes of Health; Work; chemical standard; chemical synthesis; clinical application; commercialization; drug discovery; drug efficacy; efficacy testing; enzyme activity; esterase; fluorophore; glucosidase; high throughput screening; in vitro Assay; inhibitor/antagonist; innovation; meetings; novel therapeutics; radiotracer; research clinical testing; scale up; small molecule; success; therapy development; tool

DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase II 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. established the feasibility of the technology by preparing new fluorogenic glycosidase, esterase, phosphatase, lipase and sulfatase substrates for lysosomal enzymes and demonstrated differential staining in living cells that were from normal or were of disease origin or upon induction of inhibition of lysosomal enzyme activities. In Phase II, these and additional new substrates will be assayed in vitro 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. These new systems will be validated for use in high-throughput screening for drug discovery, for use in clinical diagnostics to evaluate the occurrence and progression of disease, and for use in monitoring the effectiveness of existing or emerging therapeutic interventions for these disorders. The new substrates and the resulting detection systems will also provide innovative methods to quantitate lysosomal enzyme function and to screen for the influence of secondary drug or protein administration, making them useful medical research tools for a variety of significant biochemical and medical applications. The company has engaged the collaboration of several noted research laboratories and institutions as well as major pharmaceutical companies in this arena, who are eager to test the methods and systems in their existing clinical applications. The resulting assays and products will be marketed to the research, pharmaceutical, biotechnology and diagnostic industries. PUBLIC HEALTH RELEVANCE: The success of this project opens up enormous commercial possibilities in the fields of medical intervention in lysosomal storage disorders such as Sandhoff's disease, Tay-Sachs syndrome, Krabbi's disease and Gaucher's disease through discovery of new proteins and drugs for use in modulating lysosomal enzyme activity in these and other related diseases. These assays will also enable clinicians to determine the efficacy of current and emerging therapies as well as add a new set of clinical diagnostic assays for patient assessment. In addition, it will lead to commercial and licensable products in these areas.

描述（由申请人提供）：这个小型企业创新研究阶段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的靶向荧光底物。如果成功的话，拟议的研究将提供所需的突破，以提高发现有希望的新疗法，并调节药物以用于溶酶体储存疾病和相关医疗应用。在该项目的第一阶段，Marker Gene Technologies，Inc。通过准备新的荧光糖苷酶，酯酶，磷酸酶，磷酸酶，脂肪酶和硫酸酶底物来确定该技术的可行性，以使溶酶体酶用于溶酶体酶，并在正常或疾病中具有疾病或引诱lys的活性或lys的活性而表现出不同的活细胞。在第二阶段，这些和其他新底物将在体外测定其测量活细胞中特定和局部抑制或诱导溶酶体酶的能力，并以细胞或组织特异性方式区分单个酶活性。这些新系统将得到验证，用于用于药物发现的高通量筛查，用于评估疾病的发生和进展，并用于监测这些疾病现有或新兴治疗干预措施的有效性。新的底物和所得检测系统还将提供创新的方法来定量溶酶体酶功能并筛选二次药物或蛋白质给药的影响，从而使它们成为各种重要的生化和医学应用的有用的医学研究工具。该公司与该领域的几家著名研究实验室和机构以及主要制药公司的合作进行了合作，他们渴望在其现有临床应用中测试方法和系统。最终的测定法和产品将销售用于研究，制药，生物技术和诊断行业。 公共卫生相关性：该项目的成功在医疗干预领域为溶酶体储存障碍领域开辟了巨大的商业可能性，例如Sandhoff的疾病，Tay-Sachs综合征，Krabbi病和Gaucher病，通过发现新蛋白质和药物用于调节这些相关的这些和其他相关的疾病。这些测定还将使临床医生能够确定当前和新兴疗法的功效，并添加一组新的临床诊断测定法进行患者评估。此外，这将导致这些领域的商业和有许可产品。