Hutterer Corina, Milbradt Jens, Hamilton Stuart, Zaja Mirko, Leban Johann, Henry Christophe, Vitt Daniel, Steingruber Mirjam, Sonntag Eric, Zeitträger Isabel, Bahsi Hanife, Stamminger Thomas, Rawlinson William, Strobl Stefan, Marschall Manfred
Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg (FAU), Germany.
Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg (FAU), Germany.
Antiviral Res. 2017 Jul;143:113-121. doi: 10.1016/j.antiviral.2017.04.003. Epub 2017 Apr 9.
Infection with human cytomegalovirus (HCMV) is a serious medical problem, particularly in immunocompromised individuals and neonates. The success of (val)ganciclovir therapy is hampered by low drug compatibility and induction of viral resistance. A novel strategy of antiviral treatment is based on the exploitation of cell-directed signaling, e. g. pathways with a known relevance for carcinogenesis and tumor drug development. Here we describe a principle for putative antiviral drugs based on targeting dual-specificity tyrosine phosphorylation-regulated kinases (DYRKs). DYRKs constitute an evolutionarily conserved family of protein kinases with key roles in the control of cell proliferation and differentiation. Members of the DYRK family are capable of phosphorylating a number of substrate proteins, including regulators of the cell cycle, e.g. DYRK1B can induce cell cycle arrest, a critical step for the regulation of HCMV replication. Here we provide first evidence for a critical role of DYRKs during viral replication and the high antiviral potential of DYRK inhibitors (SC84227, SC97202 and SC97208, Harmine and AZ-191). Using established replication assays for laboratory and clinically relevant strains of HCMV, concentration-dependent profiles of inhibition were obtained. Mean inhibitory concentrations (EC50) of 0.98 ± 0.08 μM/SC84227, 0.60 ± 0.02 μM/SC97202, 6.26 ± 1.64 μM/SC97208, 0.71 ± 0.019 μM/Harmine and 0.63 ± 0.23 μM/AZ-191 were determined with HCMV strain AD169-GFP for the infection of primary human fibroblasts. A first analysis of the mode of antiviral action suggested a block of viral replication at the early-late stage of HCMV gene expression. Moreover, rhesus macaque cytomegalovirus (RhCMV), varicella-zoster virus (VZV) and herpes simplex virus (HSV-1) showed a similarly high sensitivity to these compounds. Thus, we conclude that DYRK signaling represents a promising target pathway for the development of novel anti-herpesviral strategies.
人巨细胞病毒（HCMV）感染是一个严重的医学问题，尤其是在免疫功能低下的个体和新生儿中。（缬）更昔洛韦治疗的成功受到药物相容性低和病毒耐药性诱导的阻碍。一种新的抗病毒治疗策略基于对细胞定向信号传导的利用，例如与致癌作用和肿瘤药物开发具有已知相关性的途径。在此，我们描述了一种基于靶向双特异性酪氨酸磷酸化调节激酶（DYRKs）的新型抗病毒药物的原理。DYRKs构成了一个进化上保守的蛋白激酶家族，在细胞增殖和分化的控制中起关键作用。DYRK家族成员能够磷酸化多种底物蛋白，包括细胞周期调节因子，例如DYRK1B可以诱导细胞周期停滞，这是调节HCMV复制的关键步骤。在此，我们首次提供了DYRKs在病毒复制过程中的关键作用以及DYRK抑制剂（SC84227、SC97202和SC97208、骆驼蓬碱和AZ-191）的高抗病毒潜力的证据。使用针对实验室和临床相关HCMV毒株建立的复制试验，获得了浓度依赖性抑制曲线。用HCMV毒株AD169-GFP感染原代人成纤维细胞，测定SC84227的平均抑制浓度（EC50）为0.98±0.08μM、SC97202为0.60±0.02μM、SC97208为6.26±1.64μM、骆驼蓬碱为0.71±0.019μM、AZ-191为0.63±0.23μM。对抗病毒作用模式的初步分析表明，在HCMV基因表达的早期-晚期阶段阻断了病毒复制。此外，恒河猴巨细胞病毒（RhCMV）、水痘-带状疱疹病毒（VZV）和单纯疱疹病毒（HSV-1）对这些化合物表现出同样高的敏感性。因此，我们得出结论，DYRK信号传导是开发新型抗疱疹病毒策略的一个有前景的靶标途径。
