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Over the past three decades, thrombolytic therapy for acute ischaemic stroke has evolved dramatically since the landmark National Institute of Neurological Disorders and Stroke (NINDS) trial in 1995, which formally established recombinant tissue plasminogen activator as an effective treatment for stroke. This evolution has occurred along three key dimensions: (1) an expanding repertoire of thrombolytic agents, (2) a progressive broadening of the therapeutic window and (3) organisational and technological innovations aimed at minimizing prehospital and in-hospital treatment delays.In this review, we summarise the major milestones in the clinical evidence supporting thrombolytic therapy over the past 30 years, with particular emphasis on large phase III randomised clinical trials. Our goal is to delineate the trajectory of progress in stroke thrombolysis and to provide clinicians and researchers with a clear and coherent framework for understanding the field’s past achievements and future directions.