Evaluating the Blood-Brain Barrier Bioavailability and in vivo Efficacy Potential of a Novel TAK1 Inhibitor Targeting Chronic Pain

项目摘要/摘要 慢性疼痛是一种普遍的健康问题，影响着多达1亿人，并带来经济负担 仅在美国，每年就高达5600亿美元，但治疗选择仍然有限。非甾体类非处方药 抗炎药(NSAID)是短效的，无法缓解严重的慢性疼痛，而这些 常规药物对胃肠道事件有严重的安全性担忧。此外，较新的和 更具选择性的COX-2抑制剂，如Celebrex®/Celecoxib和Vioxx®，都带有FDA的盒式警告，涉及 心脏病发作和中风的严重风险。同样，作用时间更长的中枢靶向治疗，如阿片类药物也 有较差的疗效，并产生严重的副作用，如精神状态改变、成瘾和呼吸 抑郁症。因此，有一个严重的未得到满足的需求，以替代目前的治疗方案的患者 伴随着慢性疼痛。慢性疼痛的主要驱动力之一是组织损伤或神经损伤后的炎症 由肿瘤坏死因子α等细胞因子水平升高引起，并存在正相关关系 肿瘤坏死因子水平与疼痛强度之间的关系。肿瘤坏死因子可与其位于原代细胞终末的TNFR1受体结合 传入伤害性感受器可以直接增加它们的活性，也可以刺激促炎细胞因子 产生和激活免疫细胞。我们的临床前工作已经鉴定出转化生长因子β活化蛋白1(Tak1)，一种 介导的肿瘤坏死因子促进生存/炎症反应通路中的关键信号元件。TAK1扮演着至关重要的角色 蛋白激酶介导的信号通路在促进蛋白激酶激活中的作用 慢性疼痛过程，因此已成为调节慢性疼痛和 炎症与增强的肿瘤坏死因子信号有关。我们最近对Takinib支架的发现发现了一种 高选择性、强效(IC50~2.5 nM)、口服生物可用的TAK1小分子抑制剂(Takinib类似物 HS-276)。临床前研究表明，用双亲Takinib抑制TAK1可以产生9倍的 肿瘤坏死因子水平降低，同时与促炎有关的其他细胞因子和趋化因子水平降低 回应。此外，用父母takinib抑制TAK1可以预防与疼痛相关的机械性和热热性疼痛 在炎症性疼痛模型中出现抑郁行为和浮肿。此外，父母在这方面的塔基尼治疗 模型组大鼠促炎症细胞因子和趋化因子蛋白表达减少。为了成功地达到 Takinib模拟HS-276的概念验证，该项目包括三个具体目标：目标1-评估 HS-276关于血脑屏障(BBB)生物利用度的药代动力学。目标2-确定 HS-276在尿酸单钠诱导的小鼠关节炎模型中的(治疗)镇痛作用 炎性疼痛的临床表现为机械性和热性疼痛的减少，以及炎性细胞因子的减少。 目的3-测定HS-276对链脲佐菌素诱导的小鼠急性早幼粒细胞白血病模型的治疗镇痛作用。 糖尿病神经病变。实现上述具体目标将为我们提供必要的数据 寻求NIH SBIR第二阶段申请，以资助在进入第一阶段临床试验的过程中启用IND的安全性研究。

Project Summary / Abstract Chronic pain is a prevalent health concern, affecting up to 100 million people and carrying an economic burden up to $560 billion annually in the US alone, yet treatment options remain limited. Over-the-counter nonsteroidal anti-inflammatory drugs (NSAIDs) are short-acting and fail to alleviate severe chronic pain, and these conventional drugs carry serious safety concerns regarding gastrointestinal events. In addition, the newer and more selective COX-2 inhibitors such as Celebrex®/celecoxib and Vioxx® carry FDA boxed warnings concerning serious risks of heart attack and stroke. Similarly, longer-acting centrally-targeted therapies such as opioids also have poor efficacy and produce serious side-effects regarding altered mental state, addiction, and respiratory depression. Thus, there is a serious unmet need for alternatives to current treatment options for patients suffering with chronic pain. One of the main drivers of chronic pain is inflammation following tissue injury or nerve injury caused by increased levels of cytokines such as tumor necrosis factor α (TNF), and a positive correlation exists between TNF levels and pain intensity. TNF can bind to its TNFR1 receptor located on the terminals of primary afferent nociceptors to directly increase their activity, and can also stimulate pro-inflammatory cytokine production and immune cell activation. Our preclinical work has identified TGFβ-activated kinase 1 (TAK1), a key signaling element in the mediated TNF pro-survival/inflammatory response pathway. TAK1 plays a crucial role in facilitating activation of protein kinase-mediated signaling pathways implicated in the pathogenesis of chronic pain processes, and as a result has emerged as a novel target for regulating chronic pain and inflammation linked to enhanced TNF signaling. Our recent discovery of the takinib scaffold has identified a highly selective, potent (IC50 ~2.5nM), and orally bioavailable small molecule inhibitor of TAK1 (takinib analog HS-276). Preclinical studies have demonstrated that TAK1 inhibition with parent takinib produces a 9-fold reduction in TNF levels alongside reduced levels of other cytokines and chemokines involved in pro-inflammatory responses. Also, TAK1 inhibition with parent takinib prevented mechanical and thermal heat pain, pain-related depressive behavior, and edema in a model of inflammatory pain. Furthermore, parent takinib treatment in this model reduced pro-inflammatory cytokine and chemokine protein expression. In order to successfully attain proof-of-concept for takinib analog HS-276, this project includes three Specific Aims: Aim 1 – Evaluate the pharmacokinetics of HS-276 regarding blood-brain barrier (BBB) bioavailability. Aim 2 – Determine the established (therapeutic) analgesic potential of HS-276 in a monosodium urate-induced arthritis mouse model of inflammatory pain evidenced by reduction in mechanical and thermal pain, as well as inflammatory cytokines. Aim 3 – Determine the therapeutic analgesic potential of HS-276 in a streptozotocin-induced mouse model of diabetic neuropathy. Achieving the Specific Aims above will provide will provide the necessary data for us to pursue a Phase II NIH SBIR application to fund IND-enabling safety studies en route to a Phase I clinical trial.