Integrated in vitro cytotoxicity and in silico safety profiling of glycerol-derived ethers based deep eutectic systems

Toxicity studies of Deep Eutectic Systems (DES) are of particular importance for their application in various fields, such as biomedicine and pharmaceuticals. This study evaluated the cytotoxicity of 14 DESs composed of choline chloride (ChCl) and N,N,N-triethyl-N-(2,3-dihydroxypropyl)ammonium chloride (N00Cl) as HBA, paired with structurally modified glycerol-derived ethers as HBD. Cell viability was assessed using Prestoblue and Crystal Violet assays, and protein quantification was also performed when these systems were exposed to HaCat, HepG2, and HK-2 cell lines. Furthermore, in silico ADMET predictions were performed for the individual HBA and HBD components using SwissADME and ProTox. The results obtained show that there is a dose-dependent relationship. Furthermore, analysis of the different families studied reveals that the toxicity of DESs increases with the length of the HBD chain in the case of the ChCl family, whilst an odd–even effect is observed in the N00Cl family. Additionally, the correlation between EC 50 values and XLogP3, Log Kp (sking permeation) and polarizability values was investigated, yielding direct correlations. The in silico predictions provided preliminary information for the individual components but did not fully reproduce the experimental cytotoxicity pattern of the complete DES. These findings highlight the need to assess each DES experimentally and indicate that component-based computational predictions should be interpreted as complementary to, rather thatn predictive of, the cytotoxicity of the complete euctecit systems.