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

Other compounds that inhibit known cellular targets have no effect on virus replication. (A) Cells were infected with CV-B3 or mock infected and treated with the indicated concentrations of salmeterol. At 8 h p.i., virus titers were determined by endpoint titration. Cell viability was measured by using the MTS assay. (B) Cells were infected with CV-B3 and treated with the indicated concentrations of the β2-AR antagonist ICI-118,551 (ICI) in the absence (DMSO) or presence of 5 μM formoterol (form). At 8 h p.i., virus titers were determined by a limiting-dilution assay. (C) Cells were infected with CV-B3–Rluc or mock infected and treated with various concentrations of the indicated compounds. At 6 h p.i., Renilla luciferase activity was measured. Cell viability was determined by using the MTT assay.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Other compounds that inhibit known cellular targets have no effect on virus replication. (A) Cells were infected with CV-B3 or mock infected and treated with the indicated concentrations of salmeterol. At 8 h p.i., virus titers were determined by endpoint titration. Cell viability was measured by using the MTS assay. (B) Cells were infected with CV-B3 and treated with the indicated concentrations of the β2-AR antagonist ICI-118,551 (ICI) in the absence (DMSO) or presence of 5 μM formoterol (form). At 8 h p.i., virus titers were determined by a limiting-dilution assay. (C) Cells were infected with CV-B3–Rluc or mock infected and treated with various concentrations of the indicated compounds. At 6 h p.i., Renilla luciferase activity was measured. Cell viability was determined by using the MTT assay.

Mentions: To test if the described targets of the compounds are responsible for the observed antiviral activity, we tested whether alternative compounds that act on these targets also possess antiviral activity. Cells were infected with CV-B3(-Rluc) and treated with various concentrations of the indicated compounds. Salmeterol, like formoterol, a long-acting agonist of β2 adrenergic receptors, did not inhibit virus replication up to a concentration of 33 μM, considerably higher than the EC50 of formoterol (Fig. 5A). Also, ICI-118,551, a potent antagonist of β2 adrenergic receptors, did not counteract the antiviral activity of formoterol, further supporting the idea that activation of β2 adrenergic receptors is not the mechanism of enterovirus inhibition (Fig. 5B).


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)

Other compounds that inhibit known cellular targets have no effect on virus replication. (A) Cells were infected with CV-B3 or mock infected and treated with the indicated concentrations of salmeterol. At 8 h p.i., virus titers were determined by endpoint titration. Cell viability was measured by using the MTS assay. (B) Cells were infected with CV-B3 and treated with the indicated concentrations of the β2-AR antagonist ICI-118,551 (ICI) in the absence (DMSO) or presence of 5 μM formoterol (form). At 8 h p.i., virus titers were determined by a limiting-dilution assay. (C) Cells were infected with CV-B3–Rluc or mock infected and treated with various concentrations of the indicated compounds. At 6 h p.i., Renilla luciferase activity was measured. Cell viability was determined by using the MTT assay.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Other compounds that inhibit known cellular targets have no effect on virus replication. (A) Cells were infected with CV-B3 or mock infected and treated with the indicated concentrations of salmeterol. At 8 h p.i., virus titers were determined by endpoint titration. Cell viability was measured by using the MTS assay. (B) Cells were infected with CV-B3 and treated with the indicated concentrations of the β2-AR antagonist ICI-118,551 (ICI) in the absence (DMSO) or presence of 5 μM formoterol (form). At 8 h p.i., virus titers were determined by a limiting-dilution assay. (C) Cells were infected with CV-B3–Rluc or mock infected and treated with various concentrations of the indicated compounds. At 6 h p.i., Renilla luciferase activity was measured. Cell viability was determined by using the MTT assay.
Mentions: To test if the described targets of the compounds are responsible for the observed antiviral activity, we tested whether alternative compounds that act on these targets also possess antiviral activity. Cells were infected with CV-B3(-Rluc) and treated with various concentrations of the indicated compounds. Salmeterol, like formoterol, a long-acting agonist of β2 adrenergic receptors, did not inhibit virus replication up to a concentration of 33 μM, considerably higher than the EC50 of formoterol (Fig. 5A). Also, ICI-118,551, a potent antagonist of β2 adrenergic receptors, did not counteract the antiviral activity of formoterol, further supporting the idea that activation of β2 adrenergic receptors is not the mechanism of enterovirus inhibition (Fig. 5B).

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