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Inotek Corporation正在开发一种具有有效抗炎活性的新型化合物。在这项提议中，我们提供证据表明腺苷3 （A3）受体的激动剂N6-（3-碘苄基）-腺苷-5'- n -甲基脲酰胺（IB-MECA）(1)减少多种促炎细胞因子和趋化因子的产生，(2)增强抗炎细胞因子IL-10的产生，(3)抑制一氧化氮（NO）合成酶（iNOS）的诱导异型的表达，一氧化氮合成酶是一种在结肠炎中表达并产生细胞毒性自由基一氧化氮的酶。(4)对休克和关节炎的啮齿动物模型有保护作用。Inotek目前正在寻求NIH SBIR I期资助，以确定IB-MECA作为一种新型抗结肠炎治疗药物的体内可行性。本提案的具体目的是确定一种新型A3激动剂（IB-MECA）在临床相关的啮齿动物小肠结肠炎模型中的药效学特征。我们将建立原理证明，IB-MECA是有效的大鼠小肠结肠炎模型通过结肠灌注三硝基苯磺酸在乙醇（TNBS）。IB-MECA将在随机、盲法、损伤后范式中按3个剂量水平每次灌胃给予4天，这是该模型中损伤峰值的时间点。将获得组织样本以评估粘膜损伤的组织学相关性、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表达、iNOS和硝基酪氨酸免疫反应性。证实IB-MECA在该实验模型中是一种有效的抗结肠炎药物，将证明II期申请是合理的，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），向FDA申请研究性药物和I期临床试验。拟议的商业应用：在美国，一种安全有效的炎症性肠病新疗法的市场每年将超过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.