Application of the pseudovirus-based neutralization assay in the search for new antiviral drugs

In recent years, significant progress has been made in the field of drug development, particularly due to the use of computer modeling methods. One of the key stages in the development of new antiviral drugs is testing the efficacy of promising candidates in in vitro experiments using target viruses. The application of new technologies for conducting primary screening with pseudotyped viruses simplifies research, increases its efficiency and ensures the biosafety of the conducted studies. The aim of this review is to analyze previous studies that have demonstrated the successful use of pseudovirus technology for the search of new chemotherapeutic agents against a range of RNA-containing viruses. The analysis involved the literature presented in the PubMed, Scopus, Elsevier, and Google Scholar databases as of March 1, 2025. For the search, the following keywords were used: pseudovirus, virus inhibition, antiviral drugs, RNA viruses. Pseudotyped viruses are recombinant viral particles that have the core proteins of one virus and the surface proteins of another, studied virus. The advantages of pseudovirus technology are its safety, high level of reproducibility of results, and the possibility of standardization. The lentivirus-based system was one of the first to be developed and remains one of the most in-demand. Using pseudoviruses, candidate molecules for infections caused by RNA-containing viruses, such as HIV-1, hepatitis C virus, tick-borne encephalitis virus, avian influenza viruses, and SARS-CoV-2, have been selected and studied. Most of the selected drugs act at the initial stage of the virus entry into the target cell. The examples provided illustrate the significant contribution of pseudovirus technology in dealing with serious socially significant diseases caused by RNA-containing viruses.