Medicinal plants remain central to global healthcare systems, particularly in countries such as India, which is recognized for its rich diversity of therapeutic botanicals. The genus Barleria, belonging to the family Acanthaceae, comprises over 300 species and represents the third largest genus within the family. Among these, Barleria prionitis Linn., Barleria cristata Linn, and Barleria lupulina Lindl are extensively studied due to their diverse phytochemical profiles and notable medicinal potential. This structured review synthesizes literature published between 1990 and 2025 on the medicinal relevance of the selected Barleria species. A comprehensive search was conducted using major scientific databases, including PubMed, Scopus, Web of Science, and Google Scholar. Keywords such as "phytochemistry," "traditional uses," and "pharmacological activities" guided article selection, focusing on peer-reviewed studies addressing phytochemical composition and biological activities. Phytochemical investigations reveal that these species are rich in terpenoids, iridoid glycosides, flavonoids, phenolic acids, and essential oils. These bioactive constituents demonstrate broad pharmacological properties, including antibacterial, anti-inflammatory, antioxidant, antifungal, antidiabetic, hepatoprotective, neuroprotective, immunomodulatory, cardioprotective, and anticancer effects. The diverse phytochemical spectrum underpins the wide-ranging therapeutic activities of these plants, validating their traditional uses and supporting their relevance in contemporary pharmacological research. The review highlights the ethnopharmacological importance of Barleria species and underscores their promise as sources for novel drug development, providing a foundation for future scientific and pharmaceutical exploration.