文章：

硝基基团重塑吡啶并[3,4-g]喹唑啉衍生物对胶质母细胞瘤细胞中DYRK/CLK活性及RNA剪接的影响

The Nitro Group Reshapes the Effects of Pyrido[3,4-g]quinazoline Derivatives on DYRK/CLK Activity and RNA Splicing in Glioblastoma Cells

原文发布日期：19 February 2024

DOI: 10.3390/cancers16040834

类型: Article

开放获取: 是

英文摘要：

Serine-threonine protein kinases of the DYRK and CLK families regulate a variety of vital cellular functions. In particular, these enzymes phosphorylate proteins involved in pre-mRNA splicing. Targeting splicing with pharmacological DYRK/CLK inhibitors emerged as a promising anticancer strategy. Investigation of the pyrido[3,4-g]quinazoline scaffold led to the discovery of DYRK/CLK binders with differential potency against individual enzyme isoforms. Exploring the structure–activity relationship within this chemotype, we demonstrated that two structurally close compounds, pyrido[3,4-g]quinazoline-2,10-diamine1and 10-nitro pyrido[3,4-g]quinazoline-2-amine2, differentially inhibited DYRK1-4 and CLK1-3 protein kinases in vitro. Unlike compound1, compound2efficiently inhibited DYRK3 and CLK4 isoenzymes at nanomolar concentrations. Quantum chemical calculations, docking and molecular dynamic simulations of complexes of1and2with DYRK3 and CLK4 identified a dramatic difference in electron donor-acceptor properties critical for preferential interaction of2with these targets. Subsequent transcriptome and proteome analyses of patient-derived glioblastoma (GBM) neurospheres treated with2revealed that this compound impaired CLK4 interactions with spliceosomal proteins, thereby altering RNA splicing. Importantly,2affected the genes that perform critical functions for cancer cells including DNA damage response, p53 signaling and transcription. Altogether, these results provide a mechanistic basis for the therapeutic efficacy of2previously demonstrated in in vivo GBM models.

摘要翻译： 

DYRK和CLK家族的丝氨酸-苏氨酸蛋白激酶调控多种关键细胞功能。这些酶尤其能够磷酸化参与前体mRNA剪接的蛋白质。通过药理DYRK/CLK抑制剂靶向剪接过程已成为一种有前景的抗癌策略。对吡啶并[3,4-g]喹唑啉骨架的研究发现了对特定酶亚型具有差异效力的DYRK/CLK结合剂。通过探索该化学型的构效关系，我们证明两种结构相近的化合物——吡啶并[3,4-g]喹唑啉-2,10-二胺1与10-硝基吡啶并[3,4-g]喹唑啉-2-胺2——在体外对DYRK1-4和CLK1-3蛋白激酶表现出差异抑制活性。与化合物1不同，化合物2能在纳摩尔浓度下有效抑制DYRK3和CLK4同工酶。通过量子化学计算、分子对接及分子动力学模拟对1和2与DYRK3、CLK4复合物的分析发现，两者电子供体-受体特性存在显著差异，这决定了2与这些靶点的优先结合。后续对经2处理的患者来源胶质母细胞瘤（GBM）神经球进行转录组和蛋白质组分析表明，该化合物破坏了CLK4与剪接体蛋白的相互作用，从而改变RNA剪接过程。值得注意的是，2影响了在癌细胞中执行关键功能的基因，包括DNA损伤应答、p53信号通路和转录调控。总之，这些结果为化合物2在体内GBM模型中先前展示的治疗效果提供了机制基础。

原文链接：

The Nitro Group Reshapes the Effects of Pyrido[3,4-g]quinazoline Derivatives on DYRK/CLK Activity and RNA Splicing in Glioblastoma Cells