Development of novel small molecule analgesics modulating the nNOS-NOS1AP protein-protein interaction

摘要 本申请题为“开发调节nNOS-NOS1AP蛋白的新型小分子镇痛剂-- 蛋白质相互作用“解决了治疗慢性神经病理性疼痛的更有效药物的迫切需要 影响到美国约1.16亿人。目前的止痛药，如非类固醇、类固醇、阿片类药物 加巴喷丁的类似物有文献记载，而且通常是严重的副作用，在神经性疾病中效果不佳 疼痛，并仅在有限的患者亚群中提供适当的缓解。由于它的高患病率和贫穷性 治疗方案，慢性疼痛导致的社会经济成本估计为每年5600亿至6350亿美元 我们。NMDA受体(NMDAR)的激活介导中枢神经系统的敏化，这是 与神经病理性疼痛的发展和维持有关。NMDA介导的中枢敏化 取决于受体上多蛋白质级联复合体的形成，包括突触后密度95 蛋白(PSD95)、神经元型一氧化氮合酶(NNOS)和NOS1接头蛋白(NOS1AP)。A肽 NMDAR多蛋白复合体的干扰物在临床前中风和疼痛模型中是有效的 目前正在进行缺血性中风的临床试验。靶向该复合体的小分子抑制剂具有潜在的 是有效的止痛药，没有与广泛抑制NMDARs相关的副作用。一位直接的 NMDAR复合体的下游效应是nNOS-NOS1AP。一种抑制这种复合体的化合物很可能 对神经性疼痛、中风和慢性神经系统疾病的诱发或加重有效 由过度的NMDAR活动引起。在资助的第一阶段SBIR计划中，我们的团队运行了广泛的小分子 高通量筛选nNOS-NOS1AP蛋白相互作用的抑制剂。之后 活性、选择性和初始administration-distribution-metabolism-excretion/toxicity的确认(ADME/T) 在顶端引线的研究中，我们选择了两种具有不同支架的类药物，选择性的nNOS-NOS1AP抑制剂 活体研究。两种抑制剂在疼痛模型中都是有效的。我们在其中一台上启动了一个小的化学实验 化学系列，确定区域的选择性和效力。在目前的提案中，一种传统药物 将利用药物化学的方法设计和开发新的类似物 与母体化合物比较的药代动力学特性和效力。Anagin及其研究伙伴 AMRI和印第安纳大学将通过早期的领先优化研究，至少推进一个系列。在……里面 除了提高效力和ADME/T特性外，我们还将演示最好的类似物对 细胞中的预定靶点，在两个临床前疼痛模型中验证它们的活性，并在 关键的活体行为模型。不符合我们设定标准的化合物将不会被推进。我们期待着 我们的先导化合物会比目前的止痛药有更好的治疗指标。我们有一个团队 由商业、化学、生物和活体科学专家组成，以推动这些系列朝着领先的方向发展 优化并进入临床用于治疗慢性神经病理性疼痛。

Abstract This application, “Development of novel small molecule analgesics modulating the nNOS-NOS1AP protein- protein interaction,” addresses the critical need for more effective medications to treat chronic neuropathic pain affecting ~116 million people in the United States. Current pain medications such as NSAIDS, steroids, opiates and gabapentin analogs have documented and often severe side effects, are poorly effective in neuropathic pain and provide adequate relief only in limited subsets of patients. Because of its high prevalence and poor treatment options, chronic pain results in socioeconomic costs estimated at $560-635 billion annually in the US. Activation of NMDA receptors (NMDARs) mediates central nervous system sensitization, which is implicated in the development and maintenance of neuropathic pain. NMDA-mediated central sensitization depends on formation of a multi-protein cascade complex at the receptor that includes post-synaptic density 95 protein (PSD95), neuronal nitric oxide synthase (nNOS) and NOS1 adaptor protein (NOS1AP). A peptide disruptor of the NMDAR multi-protein complex is efficacious in preclinical stroke and pain models and is currently in clinical trials for ischemic stroke. Small molecule inhibitors targeting this complex have the potential to be effective analgesics without the side effects associated with broad inhibition of NMDARs. A direct downstream effector of the NMDAR complex is nNOS-NOS1AP. A compound inhibiting this complex will likely be efficacious against neuropathic pain, stroke and chronic neurological diseases precipitated or exacerbated by excessive NMDAR activity. In the funded Phase I SBIR program, our team ran an extensive small molecule high-throughput screen to identify inhibitors of the nNOS-NOS1AP protein-protein interaction. After confirmation of activity, selectivity and initial administration-distribution-metabolism-excretion/toxicity (ADME/T) studies on the top leads, we chose two drug-like, selective nNOS-NOS1AP inhibitors with distinct scaffolds for in vivo studies. Both inhibitors are efficacious in pain models. We initiated a small chemistry effort on one chemical series, identifying regions for selectivity and potency. In the current proposal, a traditional drug medicinal chemistry approach will be used to design and develop novel analogs with improved pharmacokinetic properties and potency compared to the parent compounds. Anagin and its research partners at AMRI and Indiana University will advance at least one series through early lead optimization studies. In addition to improving potency and ADME/T properties, we will demonstrate that the best analogs are acting on the intended target in cells, validate their activity in two preclinical pain models and assess their safety profile in key behavioral in vivo models. Compounds that do not meet our set criteria will not be advanced. We anticipate that our lead compound would have a better therapeutic index than current pain medications. We have a team of business, chemistry, biology and in vivo scientific experts in place to advance these series towards lead optimization and into the clinic for the treatment of chronic neuropathic pain.