Mazzucco María B, Talarico Laura B, Vatansever Sezen, Carro Ana C, Fascio Mirta L, D'Accorso Norma B, García Cybele C, Damonte Elsa B
Laboratorio de Virología, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, UBA, Ciudad Universitaria, Piso 4, Buenos Aires, Pabellón, 1428, Argentina.
Present address: Laboratorio de Reproducción y Metabolismo, Facultad de Medicina, CEFYBO-CONICET, UBA, Buenos Aires, 1121, Argentina.
J Biomed Sci. 2015 Apr 17;22(1):29. doi: 10.1186/s12929-015-0134-2.
Dengue virus (DENV), a member of the family Flaviviridae, is at present the most widespread causative agent of a human viral disease transmitted by mosquitoes. Despite the increasing incidence of this pathogen, there are no antiviral drugs or vaccines currently available for treatment or prevention. In a previous screening assay, we identified a group of N-allyl acridones as effective virus inhibitors. Here, the antiviral activity and mode of action targeted to viral RNA replication of one of the most active DENV-2 inhibitors was further characterized.
The compound 10-allyl-7-chloro-9(10H)-acridone, designated 3b, was active to inhibit the in vitro infection of Vero cells with the four DENV serotypes, with effective concentration 50% (EC50) values in the range 12.5-27.1 μM, as determined by virus yield inhibition assays. The compound was also effective in human HeLa cells. No cytotoxicity was detected at 3b concentrations up to 1000 μM. Mechanistic studies demonstrated that virus entry into the host cell was not affected, whereas viral RNA synthesis was strongly inhibited, as quantified by real time RT-PCR. The addition of exogenous guanosine together with 3b rescued only partially the infectivity of DENV-2.
The acridone derivative 3b selectively inhibits the infection of Vero cells with the four DENV serotypes without a direct interaction with the host cell or the virion but interfering specifically with the intracellular virus multiplication. The mode of antiviral action for this acridone apparently involves the cellular enzyme inosine-monophospahe dehydrogenase together with another still unidentified target related to DENV RNA synthesis.
登革病毒（DENV）是黄病毒科的成员，目前是由蚊子传播的人类病毒性疾病最广泛的病原体。尽管这种病原体的发病率不断上升，但目前尚无抗病毒药物或疫苗可用于治疗或预防。在先前的筛选试验中，我们鉴定出一组N-烯丙基吖啶酮为有效的病毒抑制剂。在此，对最具活性的DENV-2抑制剂之一针对病毒RNA复制的抗病毒活性和作用模式进行了进一步表征。
化合物10-烯丙基-7-氯-9（10H）-吖啶酮，命名为3b，对抑制四种DENV血清型对Vero细胞的体外感染具有活性，通过病毒产量抑制试验测定，其半数有效浓度（EC50）值在12.5-27.1μM范围内。该化合物在人HeLa细胞中也有效。在高达1000μM的3b浓度下未检测到细胞毒性。机制研究表明，病毒进入宿主细胞不受影响，而病毒RNA合成受到强烈抑制，通过实时RT-PCR定量。将外源性鸟苷与3b一起添加仅部分挽救了DENV-2的感染性。
吖啶酮衍生物3b选择性抑制四种DENV血清型对Vero细胞的感染，不与宿主细胞或病毒粒子直接相互作用，但特异性干扰细胞内病毒增殖。这种吖啶酮的抗病毒作用模式显然涉及细胞酶肌苷-单磷酸脱氢酶以及另一个与DENV RNA合成相关的尚未确定的靶点。
