ADENOSINE 3 RECEPTOR AGONIST FOR TREATMENT OF COLITIS

抑制促炎细胞因子和趋化因子表达的药物可能是治疗炎症性肠病(IBD)的有效药物。总部位于马萨诸塞州的生物制药公司伊诺克公司正在开发一种具有强大抗炎活性的新型化合物。在这项建议中，我们提出了证据表明，腺苷3(A3)受体的激动剂N6-(3-碘苯基)-腺苷-5‘-N-甲基尿酸胺(IB-MECA)(1)减少多种促炎细胞因子和趋化因子的产生，(2)促进抗炎细胞因子IL-10的产生，(3)抑制诱导型一氧化氮合酶(INOS)的表达，iNOS是一种在结肠炎中表达的酶，并产生细胞毒量的自由基一氧化氮，(4)在休克和关节炎的啮齿动物模型中具有保护作用。伊诺克现在正在寻求NIH SBIR第一阶段的资金，以确定IB-MECA作为一种新型抗结肠炎治疗药物在体内的可行性。这项建议的具体目的是确定一种新的A3激动剂(IB-MECA)在临床相关的小肠结肠炎啮齿动物模型中的药效学特征。我们将建立一个原则证明，IB-MECA在一个公认的大鼠小肠结肠炎模型中是有效的，该模型是由三硝基苯磺酸在乙醇中结肠滴注(TNBS)造成的。IB-MECA将以随机、盲法、伤后模式按3个剂量水平TID每次灌胃给药，持续4天，在该模型中为损伤高峰时间点。组织标本将用于评估粘膜损伤、髓过氧化物酶活性(中性粒细胞浸润的标志)、F2-异前列腺素和丙二醛浓度(脂质过氧化的标志)、促炎细胞因子和趋化因子的表达以及诱导型一氧化氮合酶和硝基酪氨酸的免疫反应。在这个实验模型中，确认IB-MECA是一种有效的抗结肠剂将证明第二阶段的应用是合理的，以支持临床前药物测试(高级毒性测定、病理学、稳定性、药代动力学、体内灵长类动物研究)、FDA的调查性药物应用和第一阶段临床试验。拟议的商业应用：在美国，一种安全有效的治疗炎症性肠病的新疗法的市场规模将超过每年10亿美元。

Agents that inhibit pro-inflammatory cytokine and chemokine expression may be effective therapeutics for inflammatory bowel disease (IBD). Inotek Corporation, a Massachusetts-based biopharmaceutical firm, is developing a novel class of compounds with potent anti-inflammatory activity. In this proposal, we present evidence that an agonist of the adenosine 3 (A3) receptor, N6-(3-iodobenzyl)-adenosine-5'-N- methyluronamide (IB-MECA) (1) reduces the production of multiple pro- inflammatory cytokines and chemokines, (2) enhances the production of the anti-inflammatory cytokine IL-10, (3) inhibits the expression of the inducible isoform of nitric oxide (NO) synthase (iNOS), an enzyme that is expressed in colitis and produces cytotoxic amounts of the free radical nitric oxide, and (4) protects in rodent models of shock and arthritis. Inotek now seeks Phase I NIH SBIR funding to establish the in vivo feasibility of IB-MECA as a novel anti-colitic therapeutic. The Specific Aim of this proposal is to determine the pharmacodynamic profile of a novel A3 agonist (IB-MECA) in a clinically-relevant rodent model of enterocolitis. We will establish proof-of-principle that IB-MECA is effective in a well- established rat model of enterocolitis produced by colonic instillation of trinitrobenzene sulfonic acid in ethanol (TNBS). IB-MECA will be administered per gavage at 3 dose levels TID in a randomized, blinded, post-insult paradigm for 4 days, the timepoint of peak injury in this model. Tissue samples will be obtained for evaluation of histologic correlates of mucosal injury, myeloperoxidase activity (a marker of neutrophil infiltration), F2-isoprostane and malondialdehyde concentration (markers of lipoperoxidation), pro-inflammatory cytokine and chemokine expression, and iNOS and nitrotyrosine immunoreactivity. Confirmation that IB-MECA is an effective anti-colitic agent in this experimental model will justify a Phase II application to support pre-clinical pharmaceutical testing (advanced toxicity determinations, pathology, stability, pharmacokinetics, in vivo primate studies), an investigational drug application to the FDA and Phase I clinical trial. PROPOSED COMMERCIAL APPLICATIONS: The market for a safe and effective novel therapeutic for inflammatory bowel disease would is in excess of $1 billion per annum in the US.