ABSTRACT
Severe acute respiratory syndrome (SARS) is an infectious disease with a strong potential for transmission upon close personal contact and is caused by the SARS-coronavirus (CoV). However, there are no natural or synthetic compounds currently available that can inhibit SARS-CoV. We examined the inhibitory effects of 64 purified natural compounds against the activity of SARS helicase, nsP13, and the hepatitis C virus (HCV) helicase, NS3h, by conducting fluorescence resonance energy transfer (FRET)-based double-strand (ds) DNA unwinding assay or by using a colorimetry-based ATP hydrolysis assay. While none of the compounds, examined in our study inhibited the DNA unwinding activity or ATPase activity of human HCV helicase protein, we found that myricetin and scutellarein potently inhibit the SARS-CoV helicase protein in vitro by affecting the ATPase activity, but not the unwinding activity, nsP13. In addition, we observed that myricetin and scutellarein did not exhibit cytotoxicity against normal breast epithelial MCF10A cells. Our study demonstrates for the first time that selected naturally-occurring flavonoids, including myricetin and scultellarein might serve as SARS-CoV chemical inhibitors.
Subject(s)
Antiviral Agents/pharmacology , Apigenin/pharmacology , DNA Helicases/antagonists & inhibitors , Flavonoids/pharmacology , Methyltransferases/antagonists & inhibitors , Severe acute respiratory syndrome-related coronavirus/drug effects , Viral Proteins/antagonists & inhibitors , Adenosine Triphosphate/metabolism , Antiviral Agents/chemistry , Apigenin/chemistry , Breast/cytology , Breast/drug effects , Cell Line , Cell Proliferation/drug effects , Colorimetry , DNA/chemistry , DNA Helicases/chemistry , DNA Helicases/metabolism , Epithelial Cells/cytology , Epithelial Cells/drug effects , Female , Flavonoids/chemistry , Fluorescence Resonance Energy Transfer , Hepacivirus/drug effects , Hepacivirus/enzymology , Humans , Hydrolysis , Inhibitory Concentration 50 , Kinetics , RNA Helicases , Severe acute respiratory syndrome-related coronavirus/enzymology , Species Specificity , Viral Nonstructural Proteins , Viral Proteins/chemistry , Viral Proteins/metabolismABSTRACT
In this study, as a bioisosteric alternative scaffold of the antiviral aryl diketoacids (ADKs), we used 5-hydroxychromone on which two arylmethyloxy substituents were installed. The 5-hydroxychromones (5b-5g) thus prepared showed anti-HCV activity and, depending on the aromatic substituents on the 2-arylmethyloxy moiety, some of the derivatives (5b-5f) were also active against SCV. In addition, unlike the ADKs which showed selective inhibition against the helicase activity of the SCV NTPase/helicase, the 5-hydroxychromones (5b-5f) were active against both NTPase and helicase activities of the target enzyme. Among those, 3-iodobenzyloxy-substituted derivative 5e showed the most potent activity against HCV (EC(50) = 4 µM) as well as SCV (IC(50) = 4 µM for ATPase activity, 11 µM for helicase activity) and this might be used as a platform structure for future development of the multi-target or broad-spectrum antivirals.
