Rapid identification of inhibitors that interfere with poliovirus replication using a cell-based assay

Abstract

A small molecule library containing 480 known bioactive compounds was screened for antiviral activity against poliovirus (PV) using a cellular fluorescence resonance energy transfer (FRET) assay for viral protease activity. The infected reporter cells treated with the viral replication-suppressing compounds were examined via fluorescence microscope 7.5 h postinfection. Twelve molecules showed moderate to potent antiviral activity at concentrations less than 32 microM during the primary screening. Three compounds, anisomycin, linoleic acid, and lycorine, were chosen for validation. A dose-dependent cytotoxicity assay and a secondary screening using conventional plaque assay were conducted to confirm the results. The developed method can be used for rapid screening for molecules with antiviral activity.

Lycorine inhibits lipopolysaccharide-induced iNOS and COX-2 up-regulation in RAW264.7 cells through suppressing P38 and STATs activation and increases the survival rate of mice after LPS challenge

“…Previous studies have revealed that LPS-induced inflammatory mediator production by macrophages can be attenuated by isoquinoline alkaloids like berberine, and stylopine [31,32]. Lycorine is one of the most common isoquinoline alkaloids and has a variety of pharmacological activities [16][17][18][19]. Nonetheless, little information is available with respect to the molecular mechanisms underlying the anti-inflammatory effect of lycorine.…”

“…It has received much attention for its various biological effects including antitumor [16], antiviral [17], antimalarial [18], and antiinflammation [19]. Several studies have shown that lycorine induces apoptosis through regulation of the cell cycle in leukemia HL-60 cells and multiple myeloma cell line KM3 [20,21].…”

Fluoxetine Is a Potent Inhibitor of Coxsackievirus Replication

“…Lycorine hydrochloride, a plant alkaloid, was previously shown to inhibit poliovirus replication in vitro (17,43) but is known to be highly toxic to mammalian cells. Cycloheximide is also known to inhibit poliovirus replication but is a potent inhibitor of cellular translation (40).…”

Real-Time Monitoring of Human Enterovirus (HEV)-Infected Cells and Anti-HEV 3C Protease Potency by Fluorescence Resonance Energy Transfer

“…Therefore, picornaviral 3C pro is considered an attractive molecular target despite the fact that rupintrivir was not licensed following the clinical trials (8). With the development of high-throughput fluorescence microscopy and its associated image analysis system (9,15,21), this FRET-based biosensor assay represents an ideal platform adaptable for anti-3C pro drug screening. Any candidate compounds from the screening can be further validated by assaying them in HeLa-G3CwtR cells transfected with the 3C pro expression plasmid to rule out those that inhibit virus replication due to their cytotoxic properties.…”