Antiviral Activity of Rosmarinic Acid Against Four Serotypes of Dengue Virus

Abstract

The present study was undertaken to evaluate the putative antiviral activity of Rosmarinic acid (RA) against four serotypes of dengue virus (DENV). Our previous in silico binding analysis revealed that RA binds strongly to the envelope domain III (EDIII) protein of all four DENV serotypes. We employed an in vitro Biolayer Interferometry-based OCTET™ platform to study the binding interaction of RA with EDIII protein of the four DENV serotypes. Additionally, a functional plaque assay was developed to investigate the potential inhibition of infection of the four DENV serotypes. Using OCTET™, the binding interaction of RA to DENV-EDIII protein of the four DENV serotypes demonstrates interaction which can be arranged in the following order: EDIII-DENV1 (K_off value of 1.05 s^-1) > EDIII-DENV2 (K_off value of 5.63 × 10^-01 s^-1) > EDIII-DENV3 (K_off value of 4.63 × 10^-02 s^-1) > EDIII-DENV4 (K_off value of 3.53 × 10^-02 s^-1). Subsequently, the inhibiting ability of RA using plaque assay confirmed reduction in the number of plaques for all four serotypes, indicating the ability of RA not only to bind, but also to inhibit the infection of four serotypes in cell culture, while being non-toxic at the concentrations used in the study. However, the effect of RA was variable on different serotypes, demonstrating highest effect on DENV1 (EC₅₀ = 13.73 µg/mL, SI ≥ 728) followed by DENV2 (EC₅₀ = 77.74 µg/mL, SI ≥ 129), DENV3 (EC₅₀ = 244 µg/mL, SI ≥ 41) and DENV4 (EC₅₀ = 280 µg/mL, SI ≥ 36).