Liposomal nanoformulations of novel copper-based complexes exhibiting antimelanoma activity – In vitro and in vivo validation

Abstract

Melanoma stands as the most aggressive form of skin cancer. The lack of effective and safe therapies has led to the investigation of innovative strategies. The present work validates the in vitro and in vivo antimelanoma activity of new copper complexes of 8-hydroxyquinoline (8HQ) derivatives in free or liposomal forms. Firstly, the cytotoxic properties of several copper-based complexes were screened towards human (A375) and murine (B16F10) melanoma cell lines and human dermal fibroblasts or keratinocytes (HaCaT) cell lines. All the complexes presented lower IC50 values (<20 μM) than dacarbazine (DTIC) and temozolomide (TMZ), the positive controls (>80 μM). Aiming to solve low specificity against tumor cells and enhance its targetability to affected sites three metal-based complexes were selected, based on their antiproliferative properties, and incorporated in long blood circulating liposomes. One of them, di-2-(((2-morpholinoethyl)imino)methyl)quinolin-8-olCopper(II), designated as LCR35, was selected for further studies due to the highest incorporation parameters and cytotoxic properties observed. The antiproliferative activity of LCR35 was preserved after its association to liposomes. Moreover, in B16F10 cells this effect was potentiated. Furthermore, cell cycle analysis studies in A375 and B16F10 cell lines were performed to elucidate the mechanism of action of copper-based complex formulations. A cell cycle arrest at G2/M and G0/G1 phases in A375 and B16F10 cells, respectively, both in free and liposomal forms were observed. To validate the therapeutic potential of LCR35 two murine melanoma models were carried out: subcutaneous and metastatic. Pre-clinical studies demonstrated the high therapeutic effect of LCR35, especially after incorporation in liposomes, compared to control group or animals that received LCR35 Free and DTIC. Overall, in vitro and in vivo studies highlight the potential antimelanoma properties of the copper-based complex, LCR35.