Ionic liquids (ILs) exhibit cytotoxic effects on prokaryotic and eukaryotic cells. In this study, the antibacterial and antiproliferative activities of tetrahexylammonium bromide-based ILs were investigated. In order to evaluate the therapeutic potential of these ionic liquids, firstly microbiological assay using both Gram-positive and Gram-negative bacteria were conducted by employing Disk-Diffusion and 2,3,5-triphenyltetrazolium chlorine (TTC) methods to assess the antimicrobial effects. Likewise, the antitumor effects on 2D and 3D cell culture systems were assessed using the human colon cancer Caco-2 cell line and cytotoxic activity was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Alamar blue assays. Results obtained showed that [CH(CH)]N(Br) possesses an antibacterial potential, particularly in the case of two bacteria, (Gram) and (Gram) Preliminary screening of antiproliferative activity showed moderate activity, except for the concentration of 10 mM. Furthermore, regarding the effect of [CH(CH)]N(Br) on tumor cell aggregation, positive outcomes were noted. [CH(CH)]N(Br) presents promising and under-explored potential to improve antibacterial or anticancer therapies.