Purpose <p>Atopic dermatitis (AD) is a chronic, pruritic skin disorder usually treated with topical tacrolimus and corticosteroids, which are limited by adverse effects such as burning sensations and allergic reactions. We developed a new microemulgel formulation combining tacrolimus with pramoxine hydrochloride, an anesthetic that reduces pruritus by inhibiting voltage-gated sodium channels. This study aimed to optimize the formulation to augment skin retention and permeation of tacrolimus, potentially enhancing therapeutic efficacy and addressing current treatment limitations for AD.</p> Methods <p>The microemulgel formulation was optimized using a mixture design. The formulation was evaluated for physicochemical properties, in vitro drug release, ex vivo skin permeation and retention, cell viability (MTT assay), and efficacy in reducing inflammatory mediators (RT-PCR).</p> Results <p>The optimized formulation exhibited a favorable release profile, with a rapid initial release of pramoxine hydrochloride for immediate local anesthetic effects, followed by sustained release of tacrolimus for prolonged therapeutic action. Ex vivo permeation studies using porcine skin indicated enhanced permeation and increased drug retention compared to a marketed ointment. Cytotoxicity assays confirmed formulation safety, with cell viability above 90% at therapeutic concentrations. RT-PCR analysis showed a significant reduction in IL-1α expression. Stability studies at various temperatures demonstrated long-term stability and integrity, making the formulation suitable for practical use.</p> Conclusions <p>The successfully prepared microemulsion-based gel of tacrolimus and pramoxine hydrochloride demonstrated improved skin permeation and skin retention. This dual drug-loaded microemulgel offers a promising approach for AD treatment, providing immediate relief and sustained therapeutic effect, potentially improving patient complaints and treatment outcomes.</p> Graphical Abstract <p></p>