Background: Melanoma, an aggressive cancer with a poor prognosis, is difficult for early diagnosis, and there are limited drug treatments. Biologically active molecules, especially polyphenols and flavonoids, have a great therapeutic potential; however, their applications are limited by low aqueous solubility and bioavailability. Research Design and Methods: The mixture of usnic acid and curcumin was loaded into the polymer matrices based on hyaluronic acid, with the following polymeric film casting. The anticancer activity of the dual-molecule-loaded polymeric films was evaluated against lightly pigmented human melanoma SK-MEL 28 and unpigmented melanoma CVCL-7036 in comparison with the immortalized human keratinocytes HaCaT. Results: Usnic acid/curcumin-loaded biopolymer matrices demonstrated a high selective antitumor toxicity against melanoma SK-MEL 28 and CVCL-7036 cell lines with high biocompatibility with immortalized human keratinocytes HaCaT. Conclusions: Results highlight the potential of the obtained dual-molecule-loaded thin films based on hyaluronic acid as topical and safe antitumor therapy systems for local administration for the melanoma treatment. Moreover, due to the intrinsic properties of usnic acid and curcumin, and the biological activity of native hyaluronic acid, it is supposed that the obtained matrices possess the anti-inflammatory, antioxidant, antibacterial, and wound-healing activities, which are planned to be confirmed in further investigations.