Nicotiana genus represents a remarkable reservoir of bioactive compounds with excellent pharmacological potential. Accumulating evidences indicate that nicotine, flavonoids, terpenoids, and glycoalkaloids are among many specific metabolites of the Nicotiana species that have been evaluated for their anti-inflammatory, antibacterial, analgesic, antioxidant, and neuropharmacological properties. Flavonoid-mediated antioxidant protection against oxidative stress and cembranoid-based anti-inflammatory properties emphasize the importance of Nicotiana. The plant can exhibit outstanding antibacterial and antiparasitic effects through immunomodulation, enzyme inhibition, and membrane disruption. Although its promising applications concern about nicotine’s toxicity, bioactive compounds’ standardization and clinical validation still exist. This chapter summarizes medicinal chemistry of Nicotiana while highlighting the need of further studies. Nicotiana species provide unmet promise for developing novel therapies to address global health issues, such as antibiotic resistance and neurological illnesses, by integrating ethnobotany with contemporary pharmacology.