Development of Novel In Vivo Chemical Probes to Address CNS Protein Kinase Involvement in Synaptic Dysfunction
Development of Novel In Vivo Chemical Probes to Address CNS Protein Kinase Involvement in Synaptic Dysfunction
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DOI：
10.1371/journal.pone.0066226
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发表时间：
2013-06-26
期刊：
影响因子：
3.7
通讯作者：
Van Eldik, Linda J.
中科院分区：
文献类型：
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作者：
Watterson, D. Martin;Grum-Tokars, Valerie L.;Van Eldik, Linda J.
丝氨酸-苏氨酸蛋白激酶对于中枢神经系统功能至关重要，但缺乏用于体内研究的高选择性、中枢神经系统活性激酶抑制剂。此外，普遍的假设引起了人们对单一激酶抑制剂是否能够对中枢神经系统病理表现出体内功效的担忧，并且关于开发安全有效的激酶抑制剂的可行方法的争论尚未解决。因此，解决这些科学挑战至关重要，以便测试有关蛋白激酶在神经病理学进展中的假设以及体内调节其催化活性的潜力。体内调节可以减轻突触功能障碍的分子靶标的鉴定将为未来疾病缓解治疗的开发以及对细胞机制的深入了解奠定基础。临床和临床前研究表明，神经退行性疾病中的突触功能障碍与 p38 α MAPK 介导的信号级联激活之间存在重要联系。神经元和神经胶质细胞的激活还提供了异常的潜力，通过针对参与病理进展的两种不同细胞类型中的单一激酶来产生增强的反应。然而，由于缺乏适合中枢神经系统体内使用的高选择性抑制剂，靶标验证受到限制。因此，我们采用高分辨率共晶体学和药物信息学来设计和开发一种新型合成的、活性位点靶向、CNS 活性的 p38 α MAPK 抑制剂 (MW108)。通过大规模激酶组筛选、功能性 GPCR 激动剂和拮抗剂脱靶潜力分析以及细胞靶标参与评估来证明选择性。体外和体内试验表明，MW108 可改善 β-淀粉样蛋白诱导的突触和认知功能障碍。共晶分析过程中的一个偶然发现修正了有关抑制剂活性位点靶向的流行模型，提供了有助于未来激酶抑制剂设计的见解。总体而言，我们的研究提供了适合中枢神经系统研究的高选择性体内探针，并证明调节 p38 α MAPK 活性可以减轻突触功能障碍。
Serine-threonine protein kinases are critical to CNS function, yet there is a dearth of highly selective, CNS-active kinase inhibitors for in vivo investigations. Further, prevailing assumptions raise concerns about whether single kinase inhibitors can show in vivo efficacy for CNS pathologies, and debates over viable approaches to the development of safe and efficacious kinase inhibitors are unsettled. It is critical, therefore, that these scientific challenges be addressed in order to test hypotheses about protein kinases in neuropathology progression and the potential for in vivo modulation of their catalytic activity. Identification of molecular targets whose in vivo modulation can attenuate synaptic dysfunction would provide a foundation for future disease-modifying therapeutic development as well as insight into cellular mechanisms. Clinical and preclinical studies suggest a critical link between synaptic dysfunction in neurodegenerative disorders and the activation of p38 alpha MAPK mediated signaling cascades. Activation in both neurons and glia also offers the unusual potential to generate enhanced responses through targeting a single kinase in two distinct cell types involved in pathology progression. However, target validation has been limited by lack of highly selective inhibitors amenable to in vivo use in the CNS. Therefore, we employed high-resolution co-crystallography and pharmacoinformatics to design and develop a novel synthetic, active site targeted, CNS-active, p38 alpha MAPK inhibitor (MW108). Selectivity was demonstrated by large-scale kinome screens, functional GPCR agonist and antagonist analyses of off-target potential, and evaluation of cellular target engagement. In vitro and in vivo assays demonstrated that MW108 ameliorates beta-amyloid induced synaptic and cognitive dysfunction. A serendipitous discovery during co-crystallographic analyses revised prevailing models about active site targeting of inhibitors, providing insights that will facilitate future kinase inhibitor design. Overall, our studies deliver highly selective in vivo probes appropriate for CNS investigations and demonstrate that modulation of p38 alpha MAPK activity can attenuate synaptic dysfunction.