New small-molecule inhibitors effectively blocking picornavirus replication.
New small-molecule inhibitors effectively blocking picornavirus replication.
复制标题
新型小分子抑制剂可有效阻止小核糖核酸病毒复制。
DOI：
10.1128/jvi.01877-14
复制
发表时间：
2014
影响因子：
5.4
通讯作者：
Belov,GeorgeA
中科院分区：
文献类型：
--
作者：
FordSiltz,LaurenA;Viktorova,EkaterinaG;Zhang,Ben;Kouiavskaia,Diana;Dragunsky,Eugenia;Chumakov,Konstantin;Isaacs,Lyle;Belov,GeorgeA
针对小核糖核酸病毒的药物很少，这使得发现抗病毒药物成为当务之急。在这里，我们从激酶抑制剂库中鉴定并鉴定了三种化合物，这些化合物可以阻断脊髓灰质炎病毒、柯萨奇病毒B3和脑心肌炎病毒的复制。利用anin体外翻译-复制系统，我们发现这些药物抑制脊髓灰质炎病毒生命周期的不同阶段。A4(1)抑制复制复合体的形成和功能，而E5(1)和E7(2)在形成过程中最有效，但在功能阶段无效。两种化合物均未显著抑制VPg尿苷化。对E7(2)具有抗性的脊髓灰质炎病毒在3A蛋白中具有G5318A突变。该突变先前被发现赋予对环境酰亚胺样化合物的抗性，该化合物靶向病毒复制中磷脂酰肌醇4-激酶IIIβ (pi4kii β)依赖步骤。宿主蛋白募集分析表明，E7(2)降低了复制复合体上GBF1的数量；然而，pi4kii β水平保持不变。E7(2)和另一种类似环境肟的化合物GW5074干扰病毒多蛋白加工，影响3C-和2a依赖的裂解，耐药的G5318A突变部分挽救了这一缺陷。此外，E7(2)诱导病毒蛋白异常募集到膜上；因此，类环肟类化合物可能严重损害病毒多蛋白与膜的相互作用。A4(1)在脊髓灰质炎小鼠模型中显示出部分保护作用。在A4(1)或E5(1)存在的情况下，多次尝试分离耐药突变体均未成功，这表明可以在这些化合物的基础上开发有效的广谱抗病毒药物。多种小核糖核酸病毒可引发多种人类疾病，但目前只有甲型肝炎病毒和脊髓灰质炎病毒可通过疫苗接种加以控制。抗小核糖核酸病毒疗法的开发也面临着重大困难，因为这些病毒很容易对靶向病毒或细胞因子的化合物产生耐药性。在这里，我们描述了三种新型化合物，它们有效地阻断了远亲小核糖核酸病毒的复制，对细胞的毒性最小。这些化合物在尿苷化VPg引物合成后阻止病毒RNA复制。重要的是，其中两种抑制剂对耐药突变体的出现具有很强的难治性，这使它们成为进一步广谱治疗开发的有希望的候选者。其中一种化合物在脊髓灰质炎病毒模型中的评价表明，从麻痹开始部分保护。
Few drugs targeting picornaviruses are available, making the discovery of antivirals a high priority. Here, we identified and characterized three compounds from a library of kinase inhibitors that block replication of poliovirus, coxsackievirus B3, and encephalomyocarditis virus. Using anin vitrotranslation-replication system, we showed that these drugs inhibit different stages of the poliovirus life cycle. A4(1) inhibited both the formation and functioning of the replication complexes, while E5(1) and E7(2) were most effective during the formation but not the functioning step. Neither of the compounds significantly inhibited VPg uridylylation. Poliovirus resistant to E7(2) had a G5318A mutation in the 3A protein. This mutation was previously found to confer resistance to enviroxime-like compounds, which target a phosphatidylinositol 4-kinase IIIβ (PI4KIIIβ)-dependent step in viral replication. Analysis of host protein recruitment showed that E7(2) reduced the amount of GBF1 on the replication complexes; however, the level of PI4KIIIβ remained intact. E7(2) as well as another enviroxime-like compound, GW5074, interfered with viral polyprotein processing affecting both 3C- and 2A-dependent cleavages, and the resistant G5318A mutation partially rescued this defect. Moreover, E7(2) induced abnormal recruitment to membranes of the viral proteins; thus, enviroxime-like compounds likely severely compromise the interaction of the viral polyprotein with membranes. A4(1) demonstrated partial protection from paralysis in a murine model of poliomyelitis. Multiple attempts to isolate resistant mutants in the presence of A4(1) or E5(1) were unsuccessful, showing that effective broad-spectrum antivirals could be developed on the basis of these compounds.IMPORTANCEDiverse picornaviruses can trigger multiple human maladies, yet currently, only hepatitis A virus and poliovirus can be controlled with vaccination. The development of antipicornavirus therapeutics is also facing significant difficulties because these viruses readily generate resistance to compounds targeting either viral or cellular factors. Here, we describe three novel compounds that effectively block replication of distantly related picornaviruses with minimal toxicity to cells. The compounds prevent viral RNA replication after the synthesis of the uridylylated VPg primer. Importantly, two of the inhibitors are strongly refractory to the emergence of resistant mutants, making them promising candidates for further broad-spectrum therapeutic development. Evaluation of one of the compounds in anin vivomodel of poliomyelitis demonstrated partial protection from the onset of paralysis.