Cannabis sativa is a chemically diverse plant species that has attracted significant attention due to its rich profile of secondary metabolites, particularly cannabinoids. It is a rich source of bioactive compounds, particularly cannabinoids, which interact with the end cannabinoid system and exhibit diverse pharmacological effects. This study was conducted to evaluate and compile the scattered findings on the active chemical components of Cannabis sativa, with particular focus on their structures and pharmacological relevance. The aim is to provide a clearer understanding of how these compounds may contribute to therapeutic development, especially where conventional treatments remain limited. This review was undertaken to provide a comprehensive and critical synthesis of recent findings (2010–2025). This manuscript also revisits the traditional uses of C. sativa in medicine and aligns them with recent mechanistic insights from organic and medicinal chemistry. This review explores the major phytoconstituents of Cannabis sativa, including cannabinoids, terpenes, and flavonoids, and their therapeutic potential. Notably, Tetrahydrocannabinol (THC) and Cannabidiol (CBD) have demonstrated promising anti-inflammatory, neuroprotective, and analgesic properties. Additionally, cannabisderived compounds have shown antimicrobial, anticancer, and immunomodulatory activities in preclinical and clinical studies. Traditional medicine has long utilized cannabis for pain relief, appetite stimulation, and neurological disorders. Recent advancements in cannabis-based therapeutics have led to its incorporation into treatments for epilepsy, multiple sclerosis, and chronic pain management. However, a critical evaluation of current literature reveals that most data are derived from preclinical or small-scale clinical studies, with large randomized trials still lacking. Despite its therapeutic potential, challenges such as psychoactive effects, regulatory concerns, and standardization of cannabis-derived compounds remain. To fully harness its medicinal value, there is an urgent need for regulatory harmonization, development of standardized formulations, and multicenter clinical trials. The review concludes by emphasizing the need for structure-activity-guided studies, advanced synthetic analogs, and robust clinical models to translate phytochemical knowledge into reproducible therapeutic applications. DOI:10.2174/0113852728395972251207093735