The metal complex copper diethyldithiocarbamate (CuET) induces cancer cell death by inhibiting protein degradation and induces proteotoxic stress, making CuET a promising cancer therapeutic. However, no clinical formulation of CuET exists to date as the drug is insoluble in water and exhibits poor bioavailability. To develop a scalable formulation, nanoliposomal (LP) CuET was synthesized using ethanol injection as a facile one-step method that is suitable for large-scale manufacturing. The nanoparticles are monodispersed, colloidally stable, and approximately 100 nm in diameter with an encapsulation efficiency of over 80%. LP-CuET demonstrates excellent stability in plasma, minimal size change, and little drug release after six-month storage at various temperatures. Additionally, melanoma cell lines exhibit significant sensitivity to LP-CuET and cellular uptake occurs predominantly through endocytosis in YUMM 1.7 cancer cells. Intracellular drug delivery is mediated by vesicle acidification with more nanoparticles being internalized by melanoma cells compared with RAW 264.7 macrophages. Additionally, the nanoparticles preferentially accumulate in YUMM 1.7 tumors where they induce cancer cell death in vivo. The development and characterization of a stable and scalable CuET formulation illustrated in this study fulfils the requirements needed for a potent clinical grade formulation.

Czech Advanced Technology and Research Institute Palacky University Olomouc Krizkovskeho 511 8 77900 Olomouc Czech Republic

Department of Biomedical Engineering Faculty of Medicine and Health Sciences McGill University 3775 Rue University Montreal QC H3A 2B6 Canada

Division of Dermatology Department of Medicine Jewish General Hospital McGill University 3755 Cote Ste Catherine Montreal QC H3T 1E2 Canada

Division of Dermatology Department of Medicine The Ottawa Hospital University of Ottawa Ottawa ON K1H 8M5 Canada

Division of Experimental Medicine Faculty of Medicine and Health Sciences McGill University 1001 Decarie Blvd Montreal QC H4A 3J1 Canada

Faculty of Dentistry and Oral Health Sciences McGill University 3640 Rue University Montreal QC H3A 0C7 Canada

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University Olomouc Hnevotinska 1333 5 77900 Olomouc Czech Republic

Research Institute of the McGill University Health Centre 1001 Decarie Blvd Montreal QC H4A 3J1 Canada

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