The four new ligands, dialkyl esters of ( , )-propylenediamine- , '-di-(2,2'-di-(4-hydroxy-benzil))acetic acid (R - , -pddtyr·2HCl) (R = ethyl (L1), propyl (L2), butyl (L3), and pentyl (L4)) and corresponding palladium(II) complexes have been synthesized and characterized by microanalysis, infrared, H NMR and C NMR spectroscopy. cytotoxicity was evaluated using the MTT assay on four tumor cell lines, including mouse mammary (4T1) and colon (CT26), and human mammary (MDA-MD-468) and colon (HCT116), as well as non-tumor mouse mesenchymal stem cells. Using fluorescence spectroscopy were investigated the interactions of new palladium(II) complexes PdCl (R - , -pddtyr); (R = ethyl (C1), propyl (C2), butyl (C3), and pentyl (C4)) with calf thymus human serum albumin (HSA) and DNA (CT-DNA). The high values of the binding constants, , and the Stern-Volmer quenching constant, , show the good binding of all complexes for HSA and CT-DNA. The mentioned ligands and complexes were also tested on antimicrobial activity against 11 microorganisms. Testing was performed by the microdilution method, where the minimum inhibitory concentration (MMC) and the minimum microbicidal concentration (MMC) were determined.