PEG-Polymeric Nanocarriers Alleviate the Immunosuppressive Effects of Free 4-Thiazolidinone-Based Chemotherapeutics on T Lymphocyte Function and Cytokine Production

Jana Tulinska,1,* Lesya Kobylinska,2,* Miroslava Lehotska Mikusova,1 Julia Babincova,3 Natalia Mitina,4 Eva Rollerova,1 Aurelia Liskova,1 Nikola Madrova,1 Radka Alacova,3 Alexander Zaichenko,4 Roman Lesyk,5 Mira Horvathova,1 Michaela Szabova,1 Norbert Lukan,1 Sandor Vari6 1Faculty of Medicine, Slovak Medical University, Bratislava, Slovakia; 2Department of Biochemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine; 3Faculty of Public Health, Slovak Medical University, Bratislava, Slovakia; 4Department of Organic Chemistry, Lviv Polytechnic National University, Lviv, Ukraine; 5Department of Pharmaceutical, Organic and BioOrganic Chemistry Danylo Halytsky Lviv National Medical University, Lviv, Ukraine; 6International Research and Innovation in Medicine Program, Cedars - Sinai Medical Center, Los Angeles, CA, USA*These authors contributed equally to this workCorrespondence: Lesya Kobylinska, Department of Biochemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine, Email [email protected] Norbert Lukan, Faculty of Medicine, Slovak Medical University, Bratislava, Slovakia, Email [email protected]: Our study aimed to assess the effects of anticancer 4-thiazolidinone-based free water-insoluble therapeutics Les-3288 and Les-3833 and their waterborne complexes with branched PEG-containing polymeric carriers (A24-PEG550 and A24-PEG750) on immune response.Methods: Human peripheral blood was used to study in vitro lymphocyte proliferative function, leukocyte phagocytic activity and respiratory burst, and cytokine production.Results: The binding of the polymer to the anticancer drug Les-3288, which is intended to mitigate the immunosuppressive effects of the free drug on the proliferative activity of T lymphocytes and T-dependent B cells, demonstrated comparable efficacy for both A24-PEG750 and A24-PEG550 nanocarriers. Furthermore, it was observed that the drug-polymer complex significantly increased the reduced levels of IFN-γ and TNF-α resulting from free Les-3288. Conversely, the reduced levels of IL-6 and IL-4 remained unchanged. Administration of either form of Les-3288 had no effect on the phagocytic activity of monocytes, granulocytes or the respiratory burst of granulocytes. Due to the reduced cell viability and increased cytotoxicity associated with Les-3833, tenfold lower doses were selected for the immune assays. The effects of free Les-3833 on lymphocyte proliferative function resulted in significant stimulation of T-dependent B cells. The binding of Les-3833 to the smaller carrier, A24-PEG550 was found to maintain the stimulatory effect on B lymphocytes. While no effect of free Les-3833 on the granulocyte phagocytic activity was observed, binding of Les-3833 to both polymeric carriers resulted in a decrease in granulocyte phagocytic activity and respiratory burst, with no observable effect on monocytes. Monitoring of cytokine production showed no significant effect of either form of Les-3833 on the production of IFN-γ and IL-6. In the context of TNF-α and IL-4, the positive effect of polymer binding on restoring suppressed cytokine levels induced by the Les-3833 free drug was slightly more favorable for A24-PEG750.Conclusion: The drug complexation with novel PEGylated carriers is a promising way for efficient therapeutic development. Keywords: anticancer compounds, lymphocytes, phagocytic activity, respiratory burst, cytokines