title: Multiple classes of antiviral agents exhibit in vitro activity against human rhinovirus type C.
abstract: Human rhinovirus type C (HRV-C) is a newly discovered enterovirus species frequently associated with exacerbation of asthma and other acute respiratory conditions. Until recently, HRV-C could not be propagated in vitro, hampering in-depth characterization of the virus replication cycle and preventing efficient testing of antiviral agents. Herein we describe several subgenomic RNA replicon systems and a cell culture infectious model for HRV-C that can be used for antiviral screening. The replicon constructs consist of genome sequences from HRVc15, HRVc11, HRVc24, and HRVc25 strains, with the P1 capsid region replaced by a Renilla luciferase coding sequence. Following transfection of the replicon RNA into HeLa cells, the constructs produced time-dependent increases in luciferase signal that can be inhibited in a dose-dependent manner by known inhibitors of HRV replication, including the 3C protease inhibitor rupintrivir, the nucleoside analog inhibitor MK-0608, and the phosphatidylinositol 4-kinase IIIβ (PI4K-IIIβ) kinase inhibitor PIK93. Furthermore, with the exception of pleconaril and pirodavir, the other tested classes of HRV inhibitors blocked the replication of full-length HRVc15 and HRVc11 in human airway epithelial cells (HAEs) that were differentiated in the air-liquid interface, exhibiting antiviral activities similar to those observed with HRV-16. In summary, this study is the first comprehensive profiling of multiple classes of antivirals against HRV-C, and the set of newly developed quantitative HRV-C antiviral assays represent indispensable tools for the identification and evaluation of novel panserotype HRV inhibitors.
authors.affiliation: Gilead Sciences, Foster City, California, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Mello
authors.forename: Esmeralda
authors.initials: E
authors.lastname: Aguayo
authors.forename: Madeleine
authors.initials: M
authors.lastname: Rodriguez
authors.forename: Gary
authors.initials: G
authors.lastname: Lee
authors.forename: Robert
authors.initials: R
authors.lastname: Jordan
authors.forename: Tomas
authors.initials: T
authors.lastname: Cihlar
authors.forename: Gabriel
authors.initials: G
authors.lastname: Birkus
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D003139:Common Cold / Q000188:drug therapy / Q000821:virology; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D007555:Isoxazoles / Q000494:pharmacology; D010069:Oxadiazoles / Q000494:pharmacology; D010080:Oxazoles; D010649:Phenylalanine / Q000031:analogs & derivatives; D010880:Piperidines / Q000494:pharmacology; D011724:Pyridazines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D012367:RNA, Viral / Q000235:genetics; D012093:Replicon / Q000187:drug effects; D012229:Rhinovirus / Q000187:drug effects* / Q000235:genetics; D014372:Tubercidin / Q000031:analogs & derivatives / Q000494:pharmacology; D014633:Valine / Q000031:analogs & derivatives; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D007555:Isoxazoles; D010069:Oxadiazoles; D010080:Oxazoles; D010880:Piperidines; D011724:Pyridazines; D011760:Pyrrolidinones; D012367:RNA, Viral; C080202:Win 56291; D010649:Phenylalanine; C115201:pleconaril; C074589:pirodavir; D014633:Valine; D014372:Tubercidin; C518645:7-deaza-2'-C-methyladenosine; C118874:rupintrivir