Like other antimycotic imidazoles, clotrimazole interferes in the lipid synthesis of fungi and thus causes an alteration of the permeability of the cell walls. Especially when high doses are applied, there seem to be other significant mechanisms of action that are also based on enzyme inhibition. The spectrum of its efficacy includes all human pathogenic non-invasive fungi: dermatophytes (species of microsporum, trichophyton, and epidermophyton) and yeasts (candida group and Malassezia furfur). Clotrimazole is also active against Gram-positive bacteria and trichomonas. So far, there have been no indications for the development of a resistance.

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