Andrographolide and Deoxyandrographolide Inhibit Protease and IFN-Antagonist Activities of Foot-and-Mouth Disease Virus 3C(pro)

SIMPLE SUMMARY: Foot-and-Mouth disease (FMD) is a re-emerging infectious disease that poses a negative impact on livestock production and economics worldwide. It is also endemic in underdeveloped and developing countries, mostly in tropical areas. The control of this highly contagious disease requires a combination of different strategies, including the culling of infected animals, reducing animal movement, and vaccination. Although vaccination is effective, there remains a non-protective interval after immunization. Antiviral agents that can inhibit FMD virus (FMDV) could reduce the shedding of viruses in terms of quantity and duration, which could assist other control measures to contain FMD spreading. Antiviral activities of plant-based products, including andrographolides, have been demonstrated in several studies. Andrographolides are a group of phytochemical compounds derived from medicinal plants in the genus Andrographis, which are abundant in Asia, a hot spot of FMDV outbreaks. We found that andrographolides could inhibit FMDV replication by targeting a viral protease, namely 3C(pro). FMDV 3C(pro) is the main protease essential for the virus life cycle. The 3C(pro) also counteracts type I interferon, which is the frontline antiviral cytokine. We also revealed the intracellular mechanisms by which the andrographolides inhibited both protease and IFN antagonist activities of the 3C(pro). ABSTRACT: Foot-and mouth-disease (FMD) caused by the FMD virus (FMDV) is highly contagious and negatively affects livestock worldwide. The control of the disease requires a combination of measures, including vaccination; however, there is no specific treatment available. Several studies have shown that plant-derived products with antiviral properties were effective on viral diseases. Herein, antiviral activities of andrographolide (AGL), deoxyandrographolide (DAG), and neoandrographolide (NEO) against FMDV serotype A were investigated using an in vitro cell-based assay. The results showed that AGL and DAG inhibited FMDV in BHK-21 cells. The inhibitory effects of AGL and DAG were evaluated by RT-qPCR and exhibited EC50 values of 52.18 ± 0.01 µM (SI = 2.23) and 36.47 ± 0.07 µM (SI = 9.22), respectively. The intracellular protease assay revealed that AGL and DAG inhibited FMDV 3C(pro) with IC50 of 67.43 ± 0.81 and 25.58 ± 1.41 µM, respectively. Additionally, AGL and DAG significantly interfered with interferon (IFN) antagonist activity of the 3C(pro) by derepressing interferon-stimulating gene (ISGs) expression. The molecular docking confirmed that the andrographolides preferentially interacted with the 3C(pro) active site. However, NEO had no antiviral effect in any of the assays. Conclusively, AGL and DAG inhibited FMDV serotype A by interacting with the 3C(pro) and hindered its protease and IFN antagonist activities.