Antiviral activity of an N-allyl acridone against dengue virus

Journal of Biomedical Science

Elsa B Damonte³  Cybele C García³  Norma B D’Accorso⁴  Mirta L Fascio⁴  Ana C Carro³  Sezen Vatansever²  Laura B Talarico⁵  María B Mazzucco¹ 

[1] Present address: Laboratorio de Reproducción y Metabolismo, Facultad de Medicina, CEFYBO-CONICET, UBA, Buenos Aires 1121, Argentina;Graduate School of Science and Engineering, Koc University, Rumelifener yolu, Istanbul 34450, Sarıyer, Turke;IQUIBICEN-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina;CIHIDECAR (CONICET), Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, UBA, Ciudad Universitaria, Pabellón 2, Piso 3, Buenos Aires 1428, Argentina;Present address: Fundación Infant, Buenos Aires 1406, Argentina

关键词: RNA synthesis;    Acridone;    Re-emerging infection;    Antiviral;    Dengue virus;   

Others  :  1212345
DOI  :  10.1186/s12929-015-0134-2

 received in 2014-08-20, accepted in 2015-04-08,  发布年份 2015

【 摘 要 】

Background

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.

Results

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.

Conclusions

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.

【 授权许可】

   
2015 Mazzucco et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Guzmán MG, Halstead SB, Artsob H, Buchy P, Farrar J, Gubler DJ et al.. Dengue: a continuing global threat. Nat Rev Microbiol. 2010; 8:S7-S16.
• [2]Kyle JL, Harris E. Global spread and persistence of dengue. Annu Rev Microbiol. 2008; 62:71-92.
• [3]Bhatt S, Gething PW, Brady OJ, Messina JP, Farlow AW, Moyes CL et al.. The global distribution and burden of dengue. Nature. 2013; 496:504-7.
• [4]Mitka M. Dengue more prevalent than previously thought. JAMA. 2013; 309:1882.
• [5]Noble CG, Chen Y-L, Dong H, Gu F, Lim SP, Schul W et al.. Strategies for development of dengue virus inhibitors. Antiviral Res. 2010; 85:450-62.
• [6]Sampath A, Padmanabhan R. Molecular targets for flavivirus drug discovery. Antiviral Res. 2009; 81:6-15.
• [7]Borges MC, Castro LA, Da Fonseca BA. Chloroquine use improves dengue-related symptoms. Mem Inst Oswaldo Cruz. 2013; 108:596-9.
• [8]Nguyen NM, Tran CN, Phung LK, Duong KT, Huynh Hle A, Farrar J et al.. A randomized, double-blind placebo controlled trial of balapiravir, a polymerase inhibitor, in adult dengue patients. J Infect Dis. 2013; 207:1442-50.
• [9]Whitehorn J, Van Vinh CN, Truong NT, Tai LT, Van Hao N, Hien TT et al.. Lovastatin for adult patients with dengue: protocol for a randomized controlled trial. Trials. 2012; 13:203. BioMed Central Full Text
• [10]Chang J, Block TM, Guo J-T. Antiviral therapies targeting host ER alpha-glucosidases: Current status and future directions. Antiviral Res. 2013; 99:251-60.
• [11]Egloff MP, Decroly E, Malet H, Selisko B, Benarroch D, Ferron F et al.. Structural and functional analysis of methylation and 5’-RNA sequence requirements of short capped RNAs by the methyltransferase domain of dengue virus NS5. J Mol Biol. 2007; 372:723-36.
• [12]Malet H, Massé N, Selisko B, Romette J-L, Alvarez K, Guillemot JC et al.. The flavivirus polymerase as a target for drug discovery. Antiviral Res. 2008; 80:23-35.
• [13]Li H, Clum SR, You S, Ebner KE, Padmanabhan R. The serine protease and RNA-stimulated nucleoside triphosphatase and RNA helicase functional domains of dengue virus type 2 NS3 converge within a region of 20 amino acids. J Virol. 1999; 73:3108-16.
• [14]Dong H, Zhang B, Shi P-Y. Flavivirus methyltransferase: a novel antiviral target. Antiviral Res. 2008; 80:1-10.
• [15]Lescar J, Dahai L, Xu T, Sampath A, Lim SP, Canard B et al.. Towards the design of antiviral inhibitors against flaviviruses: the case for the multifunctional NS3 protein from Dengue virus as a target. Antiviral Res. 2008; 80:94-101.
• [16]Pastorino B, Nougairede A, Wurtz N, Gould E, De Lamballeire X. Role of host cell factors in flavivirus infection: Implications for pathogenesis and development of antiviral drugs. Antiviral Res. 2010; 87:281-94.
• [17]Belmont P, Bosson J, Godet T, Tiano M. Acridine and acridone derivatives, anticancer properties and synthetic methods: where are we now? Anticancer Agents Med Chem. 2007; 7:139-69.
• [18]Bastow KF. New acridone inhibitors of human herpes virus replication. Curr Drug Targets: Infect Disord. 2004; 4:323-30.
• [19]Sepúlveda CS, Fascio ML, García CC, D’Accorso NB, Damonte EB. Acridones as antiviral agents: synthesis, chemical and biological properties. Curr Med Chem. 2013; 20:2402-14.
• [20]Sasvari Z, Bach S, Blondel M, Nagy PD. Inhibition of RNA recruitment and replication of an RNA virus by acridine derivatives with known anti-prion activities. PLoS One. 2009; 4: Article ID e7376
• [21]Sepúlveda CS, Fascio ML, Mazzucco MB, Docampo Palacios ML, Pellón RF, García CC et al.. Synthesis and evaluation of N-substituted acridones as antiviral agents againts haemorrhagic fever viruses. Antivir Chem Chemother. 2008; 19:41-7.
• [22]Talarico LB, Damonte EB. Interference in dengue virus adsorption and uncoating by carrageenans. Virology. 2007; 363:473-85.
• [23]Hidari KI, Takahashi N, Arihara M, Nagaoka M, Morita K, Suzuki T. Structure and anti-dengue virus activity of sulfated polysaccharide from a marine alga. Biochem Biophys Res Commun. 2008; 376:91-5.
• [24]Kaptein SJF, De Burghgraeve T, Froeyen M, Pastorino B, Alen MMF, Mondotte JA et al.. A derivative of the antibiotic doxorubicin is a selective inhibitor of dengue and yellow fever virus replication in vitro. Antimicrob Ag Chemother. 2010; 54:5269-80.
• [25]Talarico LB, Pujol CA, Zibetti RG, Faria PC, Noseda MD, Duarte ME et al.. The antiviral activity of sulfated polysaccharides against dengue virus is dependent on virus serotype and host cell. Antiviral Res. 2005; 66:103-10.
• [26]Watterson SH, Chen P, Zhao Y, Gu HH, Dhar TG, Xiao Z et al.. Acridone-based inhibitors of inosine 5’-monophosphate dehydrogenase: discovery and SAR leading to the identification of N-(2-(6-(4-ethylpiperazin-1-yl)pyridine-3-yl)propan-2-yl)-2-fluoro-9-oxo-9,10-dihydroxyacridine-3-carboxamide (BMS-566419). J Med Chem. 2007; 50:3730-42.
• [27]Takhampunya R, Ubol S, Houng H-S, Cameron CE, Padmanabhan R. Inhibition of dengue virus replication by mycophenolic acid and ribavirin. J Gen Virol. 2006; 87:1947-52.
• [28]Leyssen P, Balzarini J, De Clercq E, Neyts J. The predominant mechanism by which ribavirin exerts its antiviral activity in vitro against flaviviruses and paramyxoviruses is mediated by inhibition of IMP dehydrogenase. J Virol. 2005; 79:1943-7.
• [29]Crabtree GW, Henderson JF. Rate-limiting steps in the interconversion of purine ribonucleotides in Ehrlich ascites tumor cells in vitro. Cancer Res. 1971; 31:985-91.
• [30]Jackson RC, Weber G, Morris HP. IMP dehydrogenase, an enzyme linked with proliferation and malignancy. Nature. 1975; 256:331-3.
• [31]Tabarrini O, Manfroni G, Fravolini A, Cecchetti V, Sabatini S, De Clercq E et al.. Synthesis and anti-BVDV activity of acridones as new potential antiviral agents. J Med Chem. 2006; 49:2621-7.
• [32]Manfroni G, Paeshuyse J, Massari S, Zanoli S, Gatto B, Maga G et al.. Inhibition of subgenomic hepatitis C virus RNA replication by acridone derivatives: identification of an NS3 helicase inhibitor. J Med Chem. 2009; 52:3354-65.
• [33]Stankiewicz-Drogon A, Palchykovska LG, Kostina VG, Alexeeva IV, Shved AD, Boguszewska-Chachulska AM. New acridone-4-carboxylic acid derivatives as potential inhibitors of Hepatitis C virus infection. Bioorg Med Chem. 2008; 16:8846-52.
• [34]Stankiewicz-Drogon A, Dorner B, Erker T, Boguszewska-Chachulska AM. Synthesis of new acridone derivatives, inhibitors of NS3 helicase, which efficiently and specifically inhibit subgenomic HCV replication. J Med Chem. 2010; 53:3117-26.
• [35]Nair V, Shu Q. Inosine monophosphate dehydrogenase as a probe in antiviral drug discovery. Antiviral Chem Chemother. 2007; 18:245-58.
• [36]Diamond MS, Zachariah M, Harris E. Mycophenolic acid inhibits dengue virus infection by preventing replication of viral RNA. Virology. 2002; 304:211-21.
• [37]Markland W, McQuaid TJ, Jain J, Kwong AD. Broad-spectrum antiviral activity of the IMP dehydrogenase inhibitor VX-497: a comparison with ribavirin and demonstration of antiviral additivity with alpha interferon. Antimicrob Agents Chemother. 2000; 44:859-66.
• [38]Nair V, Chi G, Shu Q, Julander J, Smee DF. A heterocyclic molecule with significant activity against dengue virus. Bioorg Med Chem Lett. 2009; 19:1425-7.
• [39]McDowell M, Gonzales SR, Kumarapperuma SC, Jeselnik M, Arterburn JB, Hanley KA. A novel nucleoside analog, 1-β-D-ribofuranosyl-3-ehynyl-[1,2,4]triazole (ETAR) exhibits efficacy against a broad range of flaviviruses in vitro. Antiviral Res. 2010; 87:78-80.
• [40]Qing M, Zou G, Wang Q-Y, Xu HY, Dong H, Yuan Z et al.. Characterization of dengue virus resistance to brequinar in cell culture. Antimicrob Agents Chemother. 2010; 54:3683-95.
• [41]Wang Q-Y, Bushell S, Qing M, Xu HY, Bonavia A, Nunes S et al.. Inhibition of dengue virus through suppression of host pyrimidine biosynthesis. J Virol. 2011; 85:6548-56.
• [42]Dunn MC, Knight DA, Waldman WJ. Inhibition of respiratory syncytial virus in vitro and in vivo by the immunosuppresive agent leflunomide. Antivir Ther. 2011; 16:309-17.
• [43]Marschall M, Niemann I, Kosulin K, Bootz A, Wagner S, Dobner T et al.. Assessment of drug candidates for broad-spectrum antiviral therapy targeting cellular pyrimidine biosynthesis. Antiviral Res. 2013; 100:640-8.
• [44]Kang S, Shields AR, Jupatanakul N, Dimopoulos G. Suppressing dengue-2 infection by chemical inhibition of Aedes aegypti host factors. PLoS Negl Trop Dis. 2014; 8: Article ID e3084
• [45]Sepúlveda CS, García CC, Fascio ML, D’Accorso NB, Docampo Palacios ML, Pellón RF et al.. Inhibition of Junín virus RNA synthesis by an antiviral acridone derivative. Antiviral Res. 2012; 93:16-22.