Anxiety disorders are general and psychological problems that are also linked to symptoms of depression. This study aimed to investigate the anxiolytic-like effects of Urena lobata L. (MEUL) methanolic extract in different behavioral paradigms in Swiss albino mice. For this, after an oral acute toxicity study, adult male mice were treated with MEUL (250 and 500 mg/kg, p.o.) and/or diazepam (2 mg/kg, i.p.), and subjected to a number of behavioral studies. In the open-field test, the number of square field cross, grooming, and rearing, was counted, while in the light/dark and swing test, the time spent in the dark portion and number of swings was calculated, respectively. Additionally, the phytochemical analysis was also done. Results reveal that the MEUL possesses alkaloids, glycosides, flavonoids, phenols, saponins, terpenes (including triterpenes), gums, and reducing sugars. MEUL showed a significant (p < 0.05) anxiolytic-like effect in experimental animals, where it’s dose-dependently modulated the test parameters in an open-field test. The MEUL also increased the light residence time and the number of swings in a dose-dependent manner. A dose of 500 mg/kg of MEUL caused the highest calming effect when combined with the experimental animals’ diazepam group. Taken together, findings expand an understanding of the impact of U. lobata on the central nervous system and show that this plant may be useful for the treatment of disorders associated with anxiety.