Huntington's Disease Natural Product Drug Discovery

描述（由申请人提供）：亨廷顿病（HD）是一种神经系统疾病，由编码亨廷顿蛋白的IT15基因CAG三联体重复基因突变引起。该领域的大量数据表明，htt的蛋白水解片段介导HD疾病过程，导致严重的运动障碍。该项目的目标是发现新的海洋天然产物化合物作为亨廷顿舞蹈病（HD）的潜在治疗剂。该项目填补了HD和神经退行性疾病领域的重大空白，发现了新的蛋白酶抑制剂，为未来HD治疗药物的开发提供了基础。此外，该项目利用新的海洋天然产物作为蛋白酶靶点区域药物发现的独特机会，以减少参与HD的神经毒性htt蛋白水解片段的产生。这个独特的项目将整合Vivian Hook博士在蛋白酶生物化学和神经生物学方面的专业知识，William Gerwick博士用于发现治疗药物的海洋天然产物，以及加利福尼亚大学圣地亚哥分校Albert La Spada博士的纹状体神经细胞模型，以及表达亨廷顿病突变亨廷顿蛋白（htt）的转基因小鼠。该项目计划将实施三个具有丰富经验的实验室在所需学科领域的跨学科努力，以发现亨廷顿蛋白蛋白水解的新型蛋白酶抑制剂，作为减少细胞突变htt神经毒性的手段。由于合作实验室拥有所需的专业知识，并且位于加州大学圣地亚哥分校彼此靠近，因此该项目可以快速进展。本项目的HD药物发现将在两个具体目标下完成。第一个目标是评估海洋天然产物化合物对被认为与HD有关的蛋白酶活性的抑制作用，包括半胱天蛋白酶、钙蛋白酶、溶酶体蛋白酶（组织蛋白酶L、B和D）和基质金属蛋白酶10。蓝藻属的海洋天然产物化合物是有效药物分子的丰富来源，具有独特的结构多样性。在目标2中，蓝藻海洋天然化合物将在细胞分析中测试n端htt片段的减少和对细胞死亡的保护，使用表达全长突变亨廷顿蛋白（htt）的纹状样神经元细胞和表达正常htt蛋白的对照细胞。从目标1和目标2中鉴定的有效化合物将根据其抑制能力优先考虑。这些化合物将计划用于未来的R01项目，该项目将在HD小鼠模型中对化合物进行体内测试，以有效改善HD转基因小鼠的运动损伤。结果可为HD药物开发提供候选的海洋天然药物化合物。

DESCRIPTION (provided by applicant): Huntington's Disease (HD) is a neurological disorder resulting from CAG triplet repeat genetic mutation of the IT15 gene that encodes the huntingtin protein. Extensive data in the field indicate that proteolytic fragment(s) of htt mediate the HD disease process, resulting in severe motor disturbances. The goal of this project will be to discover novel marine natural product compounds as potential therapeutic agents for Huntington's disease (HD). This project fulfills a major gap in the HD and neurodegenerative disease field for discovery of novel agents as protease inhibitors for future development of therapeutic agents for HD. Furthermore, this project uses novel marine natural products as a unique opportunity for drug discovery in the protease target area to reduce production of neurotoxic htt proteolytic fragments that participate in HD. This unique project will integrate expertise in protease biochemistry and neurobiology by Dr. Vivian Hook, marine natural products for discovery of therapeutic agents by Dr. William Gerwick, and model striatal neuronal cells, as well as transgenic mice, expressing mutant huntingtin (htt) protein of Huntington's disease by Dr. Albert La Spada at the Univ. of Calif., San Diego. The project plan will implement interdisciplinary efforts of three highly experienced laboratories in the required disciplines to discover novel protease inhibitors of huntingtin protein proteolysis as a means to reduce cellular mutant htt neurotoxicity. This project can progress at a rapid pace since the collaborating laboratories have the required expertise, and are located in close proximity to one another at UC San Diego. HD drug discovery of this project will be accomplished in two specific aims. The first aim will evaluate marine natural product compounds for inhibition of protease activities thought to be involved in HD, which includes caspase, calpain, lysosomal proteases (cathepsins L, B, and D), and matrix metalloprotease 10. Cyanobacteria algal marine natural product compounds are a rich source of potent drug molecules with unique structural diversity. In aim 2, cyanobacterial marine natural compounds will be tested in cellular assays for reduction of N-terminal htt fragments and protection from cell death, using striatal-like neuronal cells that express full-length mutant huntingtin (htt) protein, and in control cells expressing normal htt protein. Effective compounds identified from aims 1 and 2 will be prioritized by their inhibitory potencies. These compounds will be planned for a future R01 project that will conduct in vivo testing of compounds in HD mouse models for effectiveness to improve the motor impairment in HD transgenic mice. Results can indicate candidate marine natural drug compounds for HD drug development.