Liposomal Nanoformulation of a Copper-Based Complex Associated With Parpi Modulates Angiogenic Processes in Endothelial Cells

Angiogenesis is a highly regulated process and essential for the progression of solid tumors, as it ensures the continuous supply of oxygen and nutrients to metabolically dysregulated cells. This process involves degradation of the basement membrane, activation, proliferation, and migration of endothelial cells and is controlled by a balance between pro- and antiangiogenic factors, which is frequently disrupted in cancer. In this context, the tumor microenvironment plays a decisive role, as it integrates tumor, inflammatory, stromal, and endothelial cells that cooperate to induce neovascularization. Copper-based metal complexes have gained prominence due to their antitumor and antiangiogenic potential, acting through the induction of DNA damage, modulation of cell migration, and interference with molecular pathways associated with angiogenesis. The copper complex CBP-01N, combined with a PARP inhibitor (PARPi), has previously demonstrated synergistic antitumor effects, enhanced by its incorporation into a nanocolloidal liposomal system (LUV/CBP-01N-PARPi). Liposomal systems stand out as drug delivery carriers due to their biocompatibility, lipophilic nature, and compound stability, thereby optimizing biological efficacy. However, the effects of this nanoformulation on angiogenesis have not yet been described. Thus, the aim of this study was to characterize the antiangiogenic effects of the nanocolloidal system LUV/CBP-01N-PARPi by evaluating its influence on key processes, such as endothelial cell migration and neovascularization, using in vitro and ex vivo models. ECV 304 endothelial cells were used to assess cell viability by resazurin reduction and to determine the IC50. Functional assays of horizontal migration (wound healing) were performed to evaluate cell motility, while in vitro angiogenesis was investigated using the Matrigel tube formation assay. Neovascularization was also analyzed in an ex vivo murine aortic ring model. Based on the resazurin reduction assay, it was possible to determine subtoxic concentrations that did not promote significant changes in cell viability, which were then used in subsequent assays. At subtoxic concentrations, LUV/CBP-01N-PARPi significantly inhibited horizontal migration of endothelial cells and reduced in vitro tubular structure formation by approximately 40%. In the ex vivo murine aortic ring model, treatment with the nanoformulation significantly inhibited neovascularization, reducing the area occupied by vascular sprouts by approximately 80%. These findings consistently demonstrate the antiangiogenic activity of the nanoformulation, indicating that LUV/CBP-01N-PARPi presents promising antiangiogenic potential that warrants further investigation as a therapeutic strategy for tumor treatment, particularly for aggressive tumors in which angiogenesis is significantly activated.