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

摘要 该申请“开发调节 nNOS-NOS1AP 蛋白的新型小分子镇痛药 - 蛋白质相互作用”解决了对更有效的药物治疗慢性神经性疼痛的迫切需求 影响美国约 1.16 亿人。目前使用的止痛药，如非甾体抗炎药、类固醇、阿片类药物 和加巴喷丁类似物已被记录，并且通常有严重的副作用，在神经病方面效果不佳 疼痛并仅在有限的患者亚群中提供足够的缓解。由于其患病率高且贫困 治疗选择方面，慢性疼痛每年造成的社会经济成本估计为 560-6350 亿美元 美国。 NMDA 受体 (NMDAR) 的激活介导中枢神经系统敏化，这是 参与神经性疼痛的发展和维持。 NMDA介导的中枢敏化 取决于受体处多蛋白级联复合物的形成，其中包括突触后密度 95 蛋白 (PSD95)、神经元一氧化氮合酶 (nNOS) 和 NOS1 衔接蛋白 (NOS1AP)。一种肽 NMDAR 多蛋白复合物的干扰物在临床前中风和疼痛模型中有效，并且 目前正在进行缺血性中风的临床试验。针对该复合物的小分子抑制剂具有潜力 成为有效的镇痛药，且不会产生与广泛抑制 NMDAR 相关的副作用。直接一个 NMDAR 复合物的下游效应子是 nNOS-NOS1AP。抑制该复合物的化合物可能会 有效对抗神经性疼痛、中风和诱发或加剧的慢性神经系统疾病 过度的 NMDAR 活动。在资助的第一阶段 SBIR 项目中，我们的团队进行了广泛的小分子研究 高通量筛选以鉴定 nNOS-NOS1AP 蛋白质-蛋白质相互作用的抑制剂。之后 活性、选择性和初始给药-分布-代谢-排泄/毒性的确认（ADME/T） 在针对顶级先导化合物的研究中，我们选择了两种具有不同支架的药物样选择性 nNOS-NOS1AP 抑制剂 体内研究。两种抑制剂对疼痛模型均有效。我们发起了一项小型化学研究 化学系列，确定选择性和效力的区域。在目前的提案中，传统药物 药物化学方法将用于设计和开发具有改进的新型类似物 与母体化合物相比的药代动力学特性和效力。阿纳金及其研究伙伴 AMRI 和印第安纳大学将通过早期先导化合物优化研究推进至少一系列研究。在 除了提高效力和 ADME/T 特性外，我们还将证明最好的类似物正在发挥作用 细胞中的预期目标，在两个临床前疼痛模型中验证其活性并评估其安全性 关键行为体内模型。不符合我们设定标准的化合物将不会被推进。我们预计 我们的先导化合物比目前的止痛药具有更好的治疗指数。我们有一个团队 商业、化学、生物学和体内科学专家的到位，推动这些系列走向领先 优化并进入临床用于治疗慢性神经性疼痛。

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.