034 | Characterization of a ribavirin-based drug delivery system against viral nervous necrosis virus

Marianna Costa^1|2, Rosamaria Pennisi¹, Paola Trischitta¹, Federica Mastrolembo Barnà^1|2, Giuseppe Nocito³, Antonino Mazzaglia³, Maria Teresa Sciortino¹ | ¹Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Italy; ²Department of Chemistry, Biology and Biotechnology, University of Perugia, Italy; ³CNR-ISMN URT of Messina at Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Italy.

Viral nervous necrosis virus (NNV), belonging to the Nodaviridae family, is one of the major viral pathogens in marine aquaculture, especially affecting high-value commercial species such as groupers (Epinephelus spp.), causing damage to the central nervous system, behavioral alterations, and high mortality in juveniles. Despite increasing interest in pharmacological strategies to control infection, the effectiveness of antiviral agents, including ribavirin, is often limited by issues of bioavailability and tissue distribution [1,2]. In this context, drug delivery systems could be exploited to enhance the pharmacokinetic and pharmacodynamic profiles of antiviral compounds. In the present study, SSN-1 cells, which are highly permissive to NNV, were used to investigate the effects of ribavirin during NNV replication when administered as a free drug and in complex with a quaternary ammonium β-cyclodextrin soluble polymer (QABCDPS) used as a drug delivery system. We began investigating the ability of the β-cyclodextrin–based complex (QABCDPS/Ribavirin) to deliver ribavirin into SSN1 cells. To this end, we labelled the complex with a rhodamine derivative (QABCDPS/Rhod/Ribavirin) and internalization was monitored at different exposure time points by fluorescence microscopy. Moreover, selective inhibitors of the major endocytic pathways were used to study the cellular entry mechanisms used by the system. Finally, the antiviral effect of the QABCDPS/Ribavirin complex was assessed in NNV-infected SSN1 cells by RTqPCR. Altogether, results indicate that the cyclodextrinbased carrier is able to promote the ribavirin release and the following uptake within SSN1 cells, supporting its potential as an effective drugdelivery strategy to improve ribavirin performance against NNV infection.