Caffeine is a naturally occurring alkaloid compound which is widely used alone or in combination in the treatment of migraine. The short elimination half life of caffeine (3-5 h) and the relationship between its absorption from gastrointestinal tract and gastric emptying are the major obstacles toward its effective oral delivery. To surmount such limitations, transdermal proniosomal systems of caffeine were developed. A full 32 factorial design was employed using Design-Expert (R) software to study the effect of different parameters and to select the optimal proniosomal system (PNS-4). Skin irritation study and in vivo histopathological examination confirmed the safety of transdermal application of PNS-4. Radioiodination of caffeine using iodine-131 (I-131) was performed via direct electrophilic substitution reaction. Insilco docking results showed almost the same binding affnity of caffeine and I-131-Caffeine against adenosine A2A receptor. Biodistribution results showed that, transdermal I-131-Caffeine loaded PNS-4 (patch) significantly increased the residence of I-131-Caffeine in the blood with higher brain targeting than oral suspension. The obtained results proved that, PNS-4 represents a promising transdermal drug delivery system capable of overcoming challenges facing oral delivery of caffeine.