Chemical Interrogation of the Neuronal Kinome Using a Primary Cell-Based Screening Assay
Chemical Interrogation of the Neuronal Kinome Using a Primary Cell-Based Screening Assay
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DOI：
10.1021/cb300584e
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发表时间：
2013-05-01
影响因子：
4
通讯作者：
Bixby, John L.
中科院分区：
文献类型：
--
作者：
Al-Ali, Hassan;Schuerer, Stephan C.;Bixby, John L.
脊髓损伤（SCI）和创伤性脑损伤功能恢复的一个根本障碍是成人中枢神经系统缺乏足够的轴突再生。因此，需要开发能够刺激轴突生长以重新建立切断的连接的药剂。鉴于蛋白激酶在调节轴突生长中发挥的关键作用以及药物干预的潜力，小分子蛋白激酶抑制剂提供了一种有前途的治疗策略。在这里，我们报告了一种强大的基于细胞的表型测定，利用原代大鼠海马神经元，用于识别促进神经突生长的小分子激酶抑制剂。该测定方法高度可靠，适合中等通量筛选，其 Z' 因子为 0.73。筛选了一个结构多样的蛋白激酶抑制剂库，揭示了几种能够强烈且持续促进神经突生长的化合物组。在出生后皮质切片培养测定中，性能最佳的生物测定能够强劲且持续地促进轴突生长。这项研究可以作为结构活性关系 (SAR) 和其他药物发现方法的起点，以开发治疗 SCI 和相关神经病理学的药物。
A fundamental impediment to functional recovery from spinal cord injury (SCI) and traumatic brain injury is the lack of sufficient axonal regeneration in the adult central nervous system. There is thus a need to develop agents that can stimulate axon growth to re-establish severed connections. Given the critical role played by protein kinases in regulating axon growth and the potential for pharmacological intervention, small molecule protein kinase inhibitors present a promising therapeutic strategy. Here, we report a robust cell-based phenotypic assay, utilizing primary rat hippocampal neurons, for identifying small molecule kinase inhibitors that promote neurite growth. The assay is highly reliable and suitable for medium-throughput screening, as indicated by its Z'-factor of 0.73. A focused structurally diverse library of protein kinase inhibitors was screened, revealing several compound groups with the ability to strongly and consistently promote neurite growth. The best performing bioassay hit robustly and consistently promoted axon growth in a postnatal cortical slice culture assay. This study can serve as a jumping-off point for structure activity relationship (SAR) and other drug discovery approaches toward the development of drugs for treating SCI and related neurological pathologies.