Rheumatoid arthritis (RA) presents a substantial global health challenge, impacting millions worldwide, with approximately 36 million affected individuals in India alone. This study focused on formulating nine batches of niosomes (Z1-Z9) with varying compositions of cholesterol and surfactants, aiming to develop an effective transdermal delivery system for RA treatment. Evaluation parameters such as polydispersity index (PDI), vesicle size, and entrapment efficiency were meticulously assessed. Results highlighted formulations Z6 and Z8 as demonstrating superior performance compared to other batches. The optimized formulations (Z6 and Z8) exhibited high entrapment efficiencies of 84.4±1.53% and 87.6±1.46%, with particle sizes of 356±54.71 nm and 253±37.01 nm, respectively, along with acceptable PDI values. Transdermal patches were successfully developed using PEG 400 and HPMC K-100M from the Z6 and Z8 batches. Characterization of these patches revealed uniform thickness (0.332±0.001 mm and 0.328±0.001 mm), consistent weight (351.47±0.717 mg and 345.46±0.901 mg), and minimal moisture uptake (5.26±0.16 and 4.16±0.12). The Z8 batch patch emerged as particularly promising based on comprehensive evaluation criteria, suggesting its suitability for RA treatment. Furthermore, the selected optimized transdermal patches exhibited drug release kinetics following zero order, Higuchi's kinetic, and Korsemeyer-Peppas models, further supporting their potential effectiveness in delivering RA treatment.