Development of UTL-5b for Rheumatoid Arthritis: Mechanism of Action

描述(申请人提供)：类风湿性关节炎(RA)是最常见的关节炎形式之一，是一种潜在的致残性自身免疫性疾病，影响着200多万美国人。目前，DMARD(疾病修饰抗风湿药)是相对于非类固醇抗炎药(NSAID)改进的一类关节炎药物。我们已经合成并筛选了一系列创新的化合物，UTL系列，作为潜在的DMARD。其中，UTL-5b是治疗RA最有前景的化合物。UTL-5b可显著降低LD100脓毒症动物模型血中升高的肿瘤坏死因子-a水平，提高存活率。为了提供更多关于UTL-5b发挥抗关节炎作用的信息，我们建议在CIA小鼠模型中检测UTL-5b的治疗效果。这项第一阶段研究的具体目标是：(1)合成5克UTL-5b；(2)开发、分析和优化适用于拟议的体内研究的处方；(3)评估UTL-5b在胶原诱导的实验性关节炎模型中的抗关节炎活性。小鼠II型胶原诱导性关节炎(CIA)是一种实验性关节炎模型，具有类风湿性关节炎的一系列病理、免疫学和遗传学特征。大多数免疫动物都会发展成进行性炎症性关节炎，其临床特征是红斑和浮肿，受影响的爪子宽度增加通常为100%。临床评分指数已被开发来评估疾病进展到关节扭曲和脊柱炎，受影响关节的组织病理学显示滑膜炎、血管疙瘩形成以及软骨和骨侵蚀，这是由一个有效的指数来代表的。免疫学实验室发现包括对II型胶原的高抗体水平和高丙种球蛋白血症。这个模型已经被很好地建立起来，用于测试治疗类风湿性关节炎的生物和药物制剂。本研究将对UTL-5b的预防性给药和治疗性给药进行研究。除了评估UTL-5b对CIA的病理和免疫学方面的影响外，实时荧光定量PCR技术还将用于检测内毒素诱导的肿瘤坏死因子α因子(LITAF)、丝裂原激活蛋白激酶2(Map3k2)和JAK3酪氨酸蛋白激酶(JAK3)在关节炎足、淋巴结和脾中的基因活性，此外，初步的MOA研究表明UTL-5b抑制Map3K2、JAK3和LITAF，拟议的第一阶段研究的结果将在SBIR第二阶段非常有用，以设计进一步的研究方案，以清楚地确定UTL-5b的MOA。目前这项提议的最终目标是开发一种小分子肿瘤坏死因子-α调节剂，作为一种显著改进的DMARD(疾病修改抗关节炎药物)。这一阶段的研究将集中在全面的胶原蛋白诱导的关节炎动物研究上。

DESCRIPTION (provided by applicant): Rheumatoid arthritis (RA) is one of the most common forms of arthritis, a potentially crippling autoimmune disease that affects more than two million Americans. At the present time, DMARD (Disease Modifying Anti- Rheumatic Drug) is an improved class of arthritis drugs over NSAID (Non-Steroidal Anti-Inflammatory Drug). We have synthesized and screened a series of innovative compounds, UTL series, as potential DMARDs. Among them, UTL-5b is the most promising compound for RA. UTL-5b was shown to significantly lower the elevated blood level of TNF-a and increase the survival rate in an LD100 sepsis animal model. In order to develop more information concerning the potential for UTL-5b to exert anti-arthritic effects, we propose to examine the therapeutic effects of UTL-5b in a CIA mouse model. Specific aims of this Phase I study are: (1) To synthesize 5 g of UTL-5b; (2) To develop, analyze, and optimize a formulation suitable for the proposed in vivo study; (3) To evaluate the anti-arthritic activity of UTL- 5b in the collagen-induced experimental arthritis model. Type II collagen-induced arthritis (CIA) in mice is an experimental model of arthritis with a number of pathological, immunological and genetic features in common with rheumatoid arthritis. A progressive, inflammatory arthritis develops in the majority of immunized animals, which is characterized clinically by erythema and edema, with affected paw width increases of typically 100%. A clinical scoring index has been developed to assess disease progression to joint distortion and spondylitis, and histopathology of affected joints reveals synovitis, pannus formation, and cartilage and bone erosion, which is represented by a validated index. Immunological laboratory findings include high antibody levels to type II collagen, and hyper- gammaglobulinemia. This model has been well established for testing of both biological and pharmacological agents for the treatment of rheumatoid arthritis. Both prophylactic and therapeutic administration of UTL-5b will be investigated in this study. In addition to an evaluation of the influence of UTL-5b on the pathological and immunological aspects of CIA, real-time PCR techniques will be used to examine the gene activity of LPS- induced TNF alpha factor (LITAF), mitogen activated protein kinase kinase kinase 2 (Map3k2), and JAK3 tyrosine-protein kinase (JAK3) in arthritic paws, lymph nodes and spleens In addition to the preliminary MOA study, which indicated that UTL-5b suppresses Map3K2, Jak3, and LITAF, the results from the proposed Phase I studies will be very useful in SBIR Phase II in designing further study protocols to clearly identify the detailed MOA of UTL-5b. This ultimate goal of the current proposal is to develop a small molecule TNF-a modulator as a significantly improved DMARD (disease modifying anti-arthritic drug). This phase 1 study will focus on a comprehensive collagen-induced arthritis animal study.