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The study was aimed to develop a flower-based cream of Tropaeolum majus L. (Tropaeolaceae) and to evaluate its in vitro effect on Trichophyton rubrum (Arthrodermataceae) strains. The standardized extract of the flowers was obtained, then the minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined; the cream formulation was designed and elaborated and its antifungal in vitro effect was evaluated, using isoconazole 1% cream as standard. To determine significant difference in both creams, we compared the obtained inhibition halos by Student's t-test with a level of statistical significance of 0.05. The MIC was 1.8 g/mL and the MFC was 1.9 g/mL; in the study of preformulation of excipients with the standardized extract of Tropaeolum majus there were no changes. The organoleptic, physicochemical and microbiological quality controls of the antifungal cream matched with the established specifications. We could develop the antifungal cream with pharmaceutical quality; the evaluation of the in vitro effect on Trichophyton rubrum strains, with inbibition halos of 33.15 + 3.26 mm, shows that it has an antifungal effect similar with the standard cream isoconazole 1%. [ABSTRACT FROM AUTHOR]