title: Discovery of antiviral molecules for dengue: In silico search and biological evaluation.
abstract: BACKGROUND
  Dengue disease is a global disease that has no effective treatment. The dengue virus (DENV) NS2B/NS3 protease complex is a target for designing specific antivirals due to its importance in viral replication and its high degree of conservation.
  
  
  METHODS
  NS2B/NS3 protease complex structural information was employed to find small molecules that are capable of inhibiting the activity of the enzyme complex. This inhibitory activity was probed with in vitro assays using a fluorescent substrate and the complex NS2B/NS3 obtained by recombinant DNA techniques. HepG2 cells infected with dengue virus serotype 2 were used to test the activity against dengue virus replication.
  
  
  RESULTS
  A total of 210,903 small molecules from PubChem were docked in silico to the NS2B/NS3 structure (PDB: 2FOM) to find molecules that were capable of inhibiting this protein complex. Five of the best 500 leading compounds, according to their affinity values (-11.6 and -13.5 kcal/mol), were purchased. The inhibitory protease activity on the recombinant protein and antiviral assays was tested.
  
  
  CONCLUSIONS
  Chemicals CID 54681617, CID 54692801 and CID 54715399 were strong inhibitors of NS2B/NS3, with IC50 values (μM) and percentages of viral titer reductions of 19.9, 79.9%; 17.5, 69.8%; and 9.1, 73.9%, respectively. Multivariate methods applied to the molecular descriptors showed two compounds that were structurally different from other DENV inhibitors.
  
  
  CONCLUSIONS
  This discovery opens new possibilities for obtaining drug candidates against Dengue virus.
authors.affiliation: Environmental and Computational Chemistry Group, School of Pharmaceutical Sciences, University of Cartagena, Cartagena, 130014, Colombia.
authors.forename: Maria
authors.initials: M
authors.lastname: Cabarcas-Montalvo
authors.affiliation: Environmental and Computational Chemistry Group, School of Pharmaceutical Sciences, University of Cartagena, Cartagena, 130014, Colombia.
authors.forename: Wilson
authors.initials: W
authors.lastname: Maldonado-Rojas
authors.affiliation: Environmental and Computational Chemistry Group, School of Pharmaceutical Sciences, University of Cartagena, Cartagena, 130014, Colombia.
authors.forename: Diana
authors.initials: D
authors.lastname: Montes-Grajales
authors.affiliation: Environmental and Computational Chemistry Group, School of Pharmaceutical Sciences, University of Cartagena, Cartagena, 130014, Colombia.
authors.forename: Angela
authors.initials: A
authors.lastname: Bertel-Sevilla
authors.affiliation: Microbial Communication Research Group, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Irene
authors.initials: I
authors.lastname: Wagner-Döbler
authors.affiliation: Microbial Communication Research Group, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Helena
authors.initials: H
authors.lastname: Sztajer
authors.affiliation: Microbial Communication Research Group, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Reck
authors.affiliation: Laboratorio de Arbovirus, Centro de Investigaciones en Enfermedades Tropicales, Parque Tecnológico Guatiguará, Universidad Industrial de Santander, Piedecuesta, Colombia.
authors.forename: Maria
authors.initials: M
authors.lastname: Flechas-Alarcon
authors.affiliation: Laboratorio de Arbovirus, Centro de Investigaciones en Enfermedades Tropicales, Parque Tecnológico Guatiguará, Universidad Industrial de Santander, Piedecuesta, Colombia.
authors.forename: Raquel
authors.initials: R
authors.lastname: Ocazionez
authors.affiliation: Environmental and Computational Chemistry Group, School of Pharmaceutical Sciences, University of Cartagena, Cartagena, 130014, Colombia. Electronic address: joliverov@unicartagena.edu.co.
authors.forename: Jesus
authors.initials: J
authors.lastname: Olivero-Verbel
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology*; D003715:Dengue / Q000188:drug therapy* / Q000821:virology; D003716:Dengue Virus / Q000187:drug effects* / Q000201:enzymology; D055808:Drug Discovery; D056945:Hep G2 Cells; D006801:Humans; D062105:Molecular Docking Simulation; D011480:Protease Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D012697:Serine Endopeptidases / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects; D000085242:Viral Proteases
chemical_list: D000998:Antiviral Agents; D011480:Protease Inhibitors; D012697:Serine Endopeptidases; C504449:NS3 protease, dengue virus; D000085242:Viral Proteases
keywords: Dengue protease inhibitor; Inhibitory activity; In vitro inhibition; Molecular docking; NS2B/NS3 complex