ENZYME THERAPY FOR FABRY DISEASE PATIENTS

描述：法布里病是一种X连锁的先天性糖脂错误。 由溶酶体酶α缺乏引起的新陈代谢 在受影响的男性中，这会导致早期死亡，原因是 心脏、肾脏和大脑的闭塞性疾病。缺乏 足够数量的纯化酶防止了完整的 酶替代疗法治疗慢性粒细胞白血病的疗效评价 法布里病。为了获得大量的这种酶， 研究人员之前构建的杆状病毒衍生品可以产生 人类的酵素。重组酶的产量很高， 与天然的体内底物三己糖神经酰胺有活性，是 糖基化，纯化的重组酶被正常人摄取 以及细胞培养中的Fabry成纤维细胞。在此应用程序中， 研究人员建议确定重组人的生产是否 可以将酶提高到允许大规模生产的水平 适合临床试验的产品。更具体地说， 研究人员提出：(1)优化基因表达水平 产生人α-半乳糖苷酶A的重组杆状病毒； (2)将编码人α-半乳糖苷酶A的基因克隆到更多 最近构建的杆状病毒载体具有更有效的 由于存在增强子和相关载体而导致的表达 改进；(3)分析重组蛋白上存在的碳水化合物 人α-半乳糖苷酶：昆虫细胞中产生的一种酶；及(4) 使现有的纯化方案适用于PerSeptive生物系统 用于GMP生产的BIOCADHPLC型工作站。这个 酶替代疗法在高雪氏病中的成功应用 疾病，这是另一种溶酶体储存缺陷，提供了强大的 这种方法在未来的第二阶段SBIR中用于Fabry病的先例 求婚。其他有关临床终点、酶剂量和 治疗方案将在潜在的第二阶段SBIR中得到解决 求婚。 建议的商业应用：不可用

DESCRIPTION: Fabry disease is an X-linked inborn error of glycolipid metabolism caused by a deficiency of the lysosomal enzyme, alpha- galactosidase A. In affected males this leads to early death due to occlusive disease of the heart, kidney and brain. The lack of sufficient quantities of purified enzyme has prevented a complete evaluation of the potential efficacy of enzyme replacement therapy in Fabry disease. In order to obtain large quantities of this enzyme, the investigator previously constructed baculovirus derivatives that produce the human enzyme. The recombinant enzyme is produced at high levels, is active with the natural in vivo substrate, trihexosylceramide, is glycosylated, and the purified recombinant enzyme is taken up by normal and Fabry fibroblasts in cell culture. In this application, the investigator proposes to determine if production of the recombinant enzyme can be increased to levels that will permit large scale production suitable for clinical trials. More specifically, the investigator proposes: (1) to optimize the level of expression of the recombinant baculovirus that produces the human alpha-galactosidase A; (2) to clone the gene encoding the human alpha-galactosidase A into more recently constructed baculovirus vectors that have more efficient expression due to the presence of enhancers and related vector improvements; (3) to analyze the carbohydrate present on the recombinant human alpha-galactosidase A enzyme produced in insect cells; and (4) to adapt the current purification scheme to the PerSeptive Biosystems BioCAD HPLC Workstation of the type used in GMP production. The successful use of enzyme replacement therapy for patients with Gaucher's disease, which is another lysosomal storage defect, provides a strong precedent for this approach for Fabry disease in a future Phase II SBIR proposal. Other issues regarding clinical end points, enzyme dose, and treatment regimen will be addressed in a potential Phase II SBIR proposal. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE