Limits...
Screening of a Library of FDA-Approved Drugs Identifies Several Enterovirus Replication Inhibitors That Target Viral Protein 2C.

Ulferts R, de Boer SM, van der Linden L, Bauer L, Lyoo HR, Maté MJ, Lichière J, Canard B, Lelieveld D, Omta W, Egan D, Coutard B, van Kuppeveld FJ - Antimicrob. Agents Chemother. (2016)

Bottom Line: All compounds acted through the inhibition of genome replication.Mutations in the coding sequence of the coxsackievirus B3 (CV-B3) 2C protein conferred resistance to dibucaine, pirlindole, and zuclopenthixol but not formoterol, suggesting that 2C is the target for this set of compounds.Importantly, dibucaine bound to CV-B3 protein 2C in vitro, whereas binding to a 2C protein carrying the resistance mutations was reduced, providing an explanation for how resistance is acquired.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases and Immunology, Virology Division, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.

No MeSH data available.


Related in: MedlinePlus

Effects of dibucaine, pirlindole, and formoterol on selected picornaviruses in a single-cycle assay. The antipicornaviral spectra of dibucaine, pirlindole, and formoterol were analyzed in a single-cycle assay using a panel of representative picornaviruses. HeLa R19 (EV-D68, RV-A2, and RV-B14) or BGM (all other viruses) cells were infected with the indicated virus at an MOI of 10 CCID50 per cell and treated with various concentrations of dibucaine or pirlindole (0.3, 1, 3, 10, or 30 μM) or formoterol (0.1, 0.3, 1, 3, 10, or 30 μM) or mock treated. Viruses were harvested at 8 h p.i., and virus titers were determined by endpoint titration. Shown are the means and standard deviations of data from three replicates. (A) Enteroviruses; (B) cardiovirus; (C) calculated EC50 values (micromolar).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4862474&req=5

Figure 3: Effects of dibucaine, pirlindole, and formoterol on selected picornaviruses in a single-cycle assay. The antipicornaviral spectra of dibucaine, pirlindole, and formoterol were analyzed in a single-cycle assay using a panel of representative picornaviruses. HeLa R19 (EV-D68, RV-A2, and RV-B14) or BGM (all other viruses) cells were infected with the indicated virus at an MOI of 10 CCID50 per cell and treated with various concentrations of dibucaine or pirlindole (0.3, 1, 3, 10, or 30 μM) or formoterol (0.1, 0.3, 1, 3, 10, or 30 μM) or mock treated. Viruses were harvested at 8 h p.i., and virus titers were determined by endpoint titration. Shown are the means and standard deviations of data from three replicates. (A) Enteroviruses; (B) cardiovirus; (C) calculated EC50 values (micromolar).

Mentions: To analyze whether the compounds act on other related viruses, we tested their activity against representative serotypes of different enterovirus species in a single-cycle assay. The panel of viruses consisted of EV-A71 as a representative of enterovirus species A (EV-A), CV-B3 for EV-B, poliovirus Sabin 1 (EV-C), EV-D68 (EV-D), rhinovirus (RV) A2 (RV-A), and RV-B14 (RV-B). We also included encephalomyocarditis virus (EMCV) from the genus Cardiovirus. Cells were infected with the indicated viruses and treated with different concentrations of dibucaine, pirlindole, and formoterol. Due to the scarcity of zuclopenthixol, this compound could not be included in this analysis. Virus titers were determined at 8 h p.i. Pirlindole efficiently inhibited EV-D68 and CV-B3 and showed moderate activity against EV-A71 but not against any of the other viruses (Fig. 3A and B). Dibucaine was similarly active against CV-B3 and EV-D68 and was also active against EV-A71 albeit with lower potency than against CV-B3 and EV-D68 (Fig. 3A). Replication of PV, RV-A2, RV-B14 (Fig. 3A), and EMCV (Fig. 3B) was not affected by either compound at the concentrations tested. In addition to CV-B3, formoterol inhibited the replication of all other enteroviruses tested, i.e., EV-A71, RV-A2, and RV-B14 (Fig. 3A), but not the cardiovirus EMCV (Fig. 3B). This indicates that formoterol is a panenterovirus inhibitor.


Screening of a Library of FDA-Approved Drugs Identifies Several Enterovirus Replication Inhibitors That Target Viral Protein 2C.

Ulferts R, de Boer SM, van der Linden L, Bauer L, Lyoo HR, Maté MJ, Lichière J, Canard B, Lelieveld D, Omta W, Egan D, Coutard B, van Kuppeveld FJ - Antimicrob. Agents Chemother. (2016)

Effects of dibucaine, pirlindole, and formoterol on selected picornaviruses in a single-cycle assay. The antipicornaviral spectra of dibucaine, pirlindole, and formoterol were analyzed in a single-cycle assay using a panel of representative picornaviruses. HeLa R19 (EV-D68, RV-A2, and RV-B14) or BGM (all other viruses) cells were infected with the indicated virus at an MOI of 10 CCID50 per cell and treated with various concentrations of dibucaine or pirlindole (0.3, 1, 3, 10, or 30 μM) or formoterol (0.1, 0.3, 1, 3, 10, or 30 μM) or mock treated. Viruses were harvested at 8 h p.i., and virus titers were determined by endpoint titration. Shown are the means and standard deviations of data from three replicates. (A) Enteroviruses; (B) cardiovirus; (C) calculated EC50 values (micromolar).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4862474&req=5

Figure 3: Effects of dibucaine, pirlindole, and formoterol on selected picornaviruses in a single-cycle assay. The antipicornaviral spectra of dibucaine, pirlindole, and formoterol were analyzed in a single-cycle assay using a panel of representative picornaviruses. HeLa R19 (EV-D68, RV-A2, and RV-B14) or BGM (all other viruses) cells were infected with the indicated virus at an MOI of 10 CCID50 per cell and treated with various concentrations of dibucaine or pirlindole (0.3, 1, 3, 10, or 30 μM) or formoterol (0.1, 0.3, 1, 3, 10, or 30 μM) or mock treated. Viruses were harvested at 8 h p.i., and virus titers were determined by endpoint titration. Shown are the means and standard deviations of data from three replicates. (A) Enteroviruses; (B) cardiovirus; (C) calculated EC50 values (micromolar).
Mentions: To analyze whether the compounds act on other related viruses, we tested their activity against representative serotypes of different enterovirus species in a single-cycle assay. The panel of viruses consisted of EV-A71 as a representative of enterovirus species A (EV-A), CV-B3 for EV-B, poliovirus Sabin 1 (EV-C), EV-D68 (EV-D), rhinovirus (RV) A2 (RV-A), and RV-B14 (RV-B). We also included encephalomyocarditis virus (EMCV) from the genus Cardiovirus. Cells were infected with the indicated viruses and treated with different concentrations of dibucaine, pirlindole, and formoterol. Due to the scarcity of zuclopenthixol, this compound could not be included in this analysis. Virus titers were determined at 8 h p.i. Pirlindole efficiently inhibited EV-D68 and CV-B3 and showed moderate activity against EV-A71 but not against any of the other viruses (Fig. 3A and B). Dibucaine was similarly active against CV-B3 and EV-D68 and was also active against EV-A71 albeit with lower potency than against CV-B3 and EV-D68 (Fig. 3A). Replication of PV, RV-A2, RV-B14 (Fig. 3A), and EMCV (Fig. 3B) was not affected by either compound at the concentrations tested. In addition to CV-B3, formoterol inhibited the replication of all other enteroviruses tested, i.e., EV-A71, RV-A2, and RV-B14 (Fig. 3A), but not the cardiovirus EMCV (Fig. 3B). This indicates that formoterol is a panenterovirus inhibitor.

Bottom Line: All compounds acted through the inhibition of genome replication.Mutations in the coding sequence of the coxsackievirus B3 (CV-B3) 2C protein conferred resistance to dibucaine, pirlindole, and zuclopenthixol but not formoterol, suggesting that 2C is the target for this set of compounds.Importantly, dibucaine bound to CV-B3 protein 2C in vitro, whereas binding to a 2C protein carrying the resistance mutations was reduced, providing an explanation for how resistance is acquired.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Diseases and Immunology, Virology Division, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.

No MeSH data available.


Related in: MedlinePlus