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，以滑膜为目标，滑膜是关节的衬里组织。它与感染(特别是肺部)、骨质疏松症和心血管疾病的风险增加有关。全球约1%的人口患有类风湿性关节炎，它会导致关节疼痛、肿胀、僵硬和运动障碍，如果不加以控制，可能会导致不可逆转的关节损伤、严重的功能障碍和过早死亡。由于类风湿性关节炎到目前为止还没有治愈的方法，目前的大多数治疗方法都旨在延缓疾病的进展，近年来，这已经通过抗风湿病药物(DMARD)来实现。虽然抗TNF1、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，但一些问题，如注射给药途径和中和抗体的发展，导致了对可以控制RA症状的小分子化合物的研究，这些小分子化合物可以作为日常药片使用。Cognosci已经确定了一系列新的抗炎apoE模拟肽，这些肽在体外和体内抑制TNF1、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。利用前一期SBIR赠款的资金，我们证明了其中一种多肽COG133可以抑制原代人滑膜成纤维细胞中细胞因子和基质金属蛋白酶(MMP)的产生。此外，在RA的小鼠胶原诱导性关节炎(CIA)模型中，COG133减少了疾病症状并改善了组织学结果。最近，我们已经确定COG133通过与蛋白SET结合并拮抗蛋白SET的作用而发挥作用，该蛋白SET也被称为蛋白磷酸酶2A抑制物-2(I2PP2A)。COG133的S在减少治疗小鼠足爪肿胀、临床评分和软骨损伤方面的疗效，以及COG133介导的细胞因子和滑膜细胞成纤维细胞基质金属蛋白酶产生的降低，验证了SET拮抗作用和相关的PP2A激活，是开发新型RA治疗药物的真正靶点。我们现在建议开发能够高通量筛选的方法来寻找新的SET小分子抑制剂，这将允许开发一种口服活性的抗RA疗法，该疗法将通过拮抗单个靶点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，以滑膜为目标，滑膜是关节的衬里组织。它困扰着全球约1%的人口，导致关节疼痛、肿胀、僵硬和运动丧失，如果不加以控制，可能会导致不可逆转的关节损害、严重的功能障碍和过早死亡。由于类风湿性关节炎到目前为止还没有治愈的方法，目前的大多数治疗方法都旨在延缓疾病的进展，近年来，这已经通过抗风湿病药物(DMARD)来实现。虽然抗TNF1、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，但一些问题，如注射给药途径和中和抗体的发展，导致了对可以控制RA症状的小分子化合物的研究，这些小分子化合物可以作为日常药片使用。Cognosci已经确定了一系列新的抗炎肽，可以抑制RA小鼠模型的疾病症状和关节损伤。这些数据证实，一种模仿较大多肽的小化合物可以作为类风湿性关节炎的日常治疗药物，可以作为药片而不是注射使用。在这方面，我们建议开发有助于识别这些新的药物线索的分析方法，这些药物可以用于开发新的抗RA疗法。

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.