Introduction: The emergence of resistant strains of viruses, recurrence of diseases, ineffectiveness of known drugs against the latent form of infection, encourage scientists to seek new means of prevention and treatment of such infection. Incorporation of the fluorine atom into compounds can impact their solubility and lipophilicity, and affect their biological potency. The aim of this study was to elucidate the impact of fluorine-containing thiosugars on herpetic and adenoviral infections. Methodology: The toxicity of trifluoromethylthiolane derivatives: 2-hydroxy-2-trifluoromethylthiolane (10S52) or 3-hydroxy-2-(hydroxymethyl)-trifluoromethylthiolane (SBIO6) was evaluated using changes in cell viability and metabolic activity. The antiviral activity against herpes simplex virus type 1 (HSV-1) and human adenovirus type 2 (HAdV-2) was assessed using a virus yield reduction assay. Results: Some cytotoxic effect of SBIO6 and 10S52 compounds on BHK-21and Hep-2 cells was detected, CC₅₀ values were in ranging 161-670 µg/ml. It was found that compound 10S52 significantly inhibited HSV-1 reproduction, reducing the virus titer obtained de novo by 1.7 lg. Such activity indicates that virus offspring are formed but virus particles are not complete and they are not able to cause an infection process. Whereas for adenoviral infection such effect was not detected. Conclusion: Thus, the 2-hydroxy-2-trifluoromethylthiolane may be a potential tool for the development of agents for the treatment of HSV-1 infections. Funding: This work was supported by scientific project of NAS of Ukraine “Patterns of influence of chemical, natural and physical virus-inactivating agents on biological processes in the virus-cell system”.