With the increasing development of metallopharmaceuticals, coordination compounds become viable alternatives for therapeutic uses. Despite the importance of platinum derivatives in this area, first-row transition metals complexes are welcome due to their characteristics. Vanadium is a promising metal in this context, as it has a range of compounds with different biological applications, including anticancer therapeutic effects. In this effort, the study of interactions between coordination compounds with deoxyribonucleic acid and with human serum albumin is fundamental. In this way, ten iminic ligands were synthesized by condensing p-substituted aromatic benzohydrazides (OH, CH3, H, NO2, and NH2) with salicylaldehyde (L1As-L5As) or pyridoxal hydrochloride (L1P-L5P). These ligands have characteristics that allow the tridentate coordination of vanadium cations, leading to the formation of ten vanadium(V) complexes (C1As-C5As and C1P-C5P) with different structural features, all characterized by single-crystal X-ray diffraction, UV–Vis and infrared spectroscopies, and cyclic voltammetry. In addition, the complexes were tested for their interactions with calf thymus deoxyribonucleic acid and human serum albumin by spectroscopic assays and molecular docking calculations. These new results can contribute to further research and provide different ways to design new vanadium complexes with biological applications.