Alopecia areata (AA) is a complex autoimmune disease that has a negative impact on the psychological well-being of patients. AA is associated with T-cells activation and cytokines release leading to collapse of immune privilege of hair follicles (HF). Tofacitinib, a JAK 1&3 inhibitor, exhibited effectiveness in AA treatment. The aim of this study was to develop gelatin-coated transferosomes (GLTS) to deliver tofactinib specifically to the HF to enhance the treatment of AA. GLTS were evaluated for ex vivo skin permeation, localization in skin layers by the tape stripping technique and Confocal microscopy. Finally, GLTS gel was applied topically for the treatment of AA patients, where seven AA patients with recalcitrant lesions (5 males and 2 females) were included in this study, then they were evaluated clinically and dermoscopically to assess the efficacy of treatment. GLTS of size 223.23 ± 16.43 nm, exhibited the highest HF localization by tape stripping (7.8561 ± 0.77 μg), and the highest mean fluorescence intensity in HF (84.63 ± 7.98 rfu). Additionally, hair regrowth in all AA patients was observed after 12 weeks with up to 80 % improvement. The present work proposed effective formulations for HF targeting of tofacitinib and proved enhanced clinical efficacy in recalcitrant AA patients with positive feedback.