High Throughput Screen for Novel Anti-Rheumatic Compounds

描述（由申请人提供）： 类风湿性关节炎（RA）是一种慢性、全身性、炎症性自身免疫性疾病，其靶点为滑膜，即关节内衬的组织。它与感染（特别是肺部感染）、骨质疏松症和心血管疾病的风险增加有关。RA影响全球约1%的人口，导致关节疼痛、肿胀、僵硬和运动丧失，如果不加控制，可导致不可逆的关节损伤、严重的功能障碍和过早死亡。由于迄今为止RA还没有治愈方法，因此目前大多数疗法旨在减缓疾病进展，近年来，这已经通过疾病修饰抗风湿药物（DMARD）来实现。虽然抗TNF 1、抗IL-12和最近的抗IL-6 DMARD化合物已经成功地治疗了RA患者，但许多问题，如注射递送途径和中和抗体的开发，已经导致寻找当作为每日药丸递送时可以控制RA症状的小分子化合物。Cognosci已经鉴定了一系列新型抗炎apoE模拟肽，其在体外和体内抑制TNF 1、IL-12、IL-6和一氧化氮的产生，因此具有RA治疗所需的许多特性。利用以前的第一阶段SBIR资助的资金，我们证明了这些肽之一，COG 133，抑制细胞因子和基质金属蛋白酶（MMP）酶在原代人滑膜成纤维细胞的生产。此外，在RA的鼠胶原诱导的关节炎（CIA）模型中，COG 133减轻了疾病症状并改善了组织学结果。最近，我们已经确定COG 133通过结合并拮抗蛋白质SET（也称为蛋白质磷酸酶2A（I2 PP 2A）的抑制剂-2）的作用而起作用。C 0 G 133在减少治疗小鼠的爪中的肿胀、临床评分和软骨损伤方面的功效，加上C 0 G 133介导的从滑膜细胞成纤维细胞产生的细胞因子和MMP的降低，验证了SET拮抗作用和相关的PP 2A活化，作为开发新型RA治疗剂的真正靶标。我们现在建议开发能够进行高通量筛选的测定，以鉴定SET的新型小分子抑制剂，其将允许开发口服活性抗RA疗法，其将通过拮抗单一靶标来抑制多种炎症途径。1 公共卫生相关性： 类风湿性关节炎（RA）是一种慢性、全身性、炎症性自身免疫性疾病，其靶点为滑膜，即关节内衬的组织。它困扰着全世界约1%的人口，并导致关节疼痛、肿胀、僵硬和运动丧失，如果不加控制，可导致不可逆的关节损伤、严重的功能障碍和过早死亡。由于迄今为止RA还没有治愈方法，因此目前大多数疗法旨在减缓疾病进展，近年来，这已经通过疾病修饰抗风湿药物（DMARD）来实现。虽然抗TNF 1、抗IL-12和最近的抗IL-6 DMARD化合物已经成功地治疗了RA患者，但许多问题，如注射递送途径和中和抗体的开发，已经导致寻找当作为每日药丸递送时可以控制RA症状的小分子化合物。Cognosci已经在RA小鼠模型中鉴定出一系列新的抗炎肽，这些肽可以抑制减少疾病症状和关节损伤。这些数据证实，一种模拟较大肽的小化合物可以作为RA的日常治疗药物，而不是注射剂。在这方面，我们建议开发检测，这将有助于确定这些新的药物线索，可用于开发一种新的抗RA治疗。

DESCRIPTION (provided by applicant): Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory autoimmune disease targeting the synovium, the tissue that lines the joints. It is associated with increased risk of infection (pulmonary in particular), osteoporosis, and cardiovascular disease. Afflicting about 1 % of the population worldwide, RA causes pain, swelling, stiffness, and loss of motion in the joints, and, if unchecked, can cause irreversible joint damage, severe functional impairment, and premature death. Since RA thus far has no cure, most current therapies aim to slow disease progression and in recent years this has been accomplished with Disease Modifying Anti- Rheumatic Drugs (DMARD). While the anti-TNF1, anti-IL-12, and more recently anti-IL-6 DMARD compounds have successfully treated RA patients, a number of problems such as an injectable route of delivery and the development of neutralizing antibodies have lead to the search for small molecule compounds that can control RA symptoms when delivered as a daily pill. Cognosci has identified a novel series of anti-inflammatory apoE-mimetic peptides that suppress production of TNF1, IL-12, IL-6, and nitric oxide in vitro and in vivo, and thus have many of the properties necessary for an RA therapy. Using funds from a previous Phase I SBIR grant, we demonstrated that one of these peptides, COG133, inhibited production of cytokines and matrix metalloprotease (MMP) enzymes in primary human synovial fibroblasts. Furthermore, COG133 reduced disease symptoms and improved histological outcome in the murine collagen induced arthritis (CIA) model of RA. Recently, we have determined that COG133 acts by binding to and antagonizing the actions of the protein SET, also known as Inhibitor-2 of Protein Phosphatase 2A (I2PP2A). COG133's efficacy at reducing swelling, clinical scores, and cartilage damage in the paws of treated mice, coupled with COG133-mediated lowering of cytokine and MMP production from synovial cell fibroblasts, validates SET antagonism, and the associated PP2A activation, as a bona fide target for the development of novel RA therapeutics. We now propose to develop assays to enable high throughput screening to identify novel small molecule inhibitors of SET that will allow for development of an orally active anti-RA therapy that will inhibit multiple inflammatory pathways by antagonism of a single target. 1 PUBLIC HEALTH RELEVANCE: Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory autoimmune disease targeting the synovium, the tissue that lines the joints. It afflicts about 1 % of the population worldwide and causes pain, swelling, stiffness, and loss of motion in the joints, and, if unchecked, can cause irreversible joint damage, severe functional impairment, and premature death. Since RA thus far has no cure, most current therapies aim to slow disease progression and in recent years this has been accomplished with Disease Modifying Anti- Rheumatic Drugs (DMARD). While the anti-TNF1, anti-IL-12, and more recently anti-IL-6 DMARD compounds have successfully treated RA patients, a number of problems such as an injectable route of delivery and the development of neutralizing antibodies have lead to the search for small molecule compounds that can control RA symptoms when delivered as a daily pill. Cognosci has identified a novel series of anti-inflammatory peptides that suppress reduce disease symptoms and joint damage in an mouse model of RA. These data validate that a small compound that mimics the larger peptides could work as a daily therapy for RA that would be taken as a pill rather than injections. In this we propose to develop assays that will help to identify these novel drug leads that can be used to develop a new anti-RA therapy.