Abstract

Objective Oral squamous cell carcinoma (OSCC), a common subtype of head and neck cancer (HNSC), impairs essential functions such as breathing, swallowing, and speech. This study aimed to evaluate the therapeutic potential of targeting mitochondrial pathways via inhibition of Voltage-Dependent Anionic Channel 1 (VDAC1) using itraconazole (ITRA) and to assess the synergistic effect of combining VDAC1 inhibition with autophagy inhibition using hydroxychloroquine (HCQ).Methods The effects of ITRA, HCQ, and their combinations with cisplatin were tested in various HNSC models, including monolayer cultures, hypoxic conditions, 3D spheroids, and an in vivo xenograft model. The study also examined the role of the VDAC1-Hexokinase-II complex and the impact of autophagy under metabolic stress.Results ITRA alone showed limited cytotoxicity, but its combination with HCQ significantly enhanced antitumoral activity. Synergistic effects were observed with HCQ + ITRA, HCQ + cisplatin, and ITRA + cisplatin in all in vitro models. In vivo, HCQ alone and in combination with ITRA reduced tumor growth without adverse effects.Conclusions Taken together, these findings support itraconazole as a promising candidate for further in vivo and clinical investigation in selected oral cancers, including HNSCC and OSCC. Nevertheless, the definition of clinical treatment schedules will require additional comprehensive pharmacokinetic, toxicological, and clinical studies.