Artemisia vulgaris L. (Mugwort or Afsantin) has been used to treat various diseases since ancient times by the inhabitants of Himalayan region-Pakistan. Methanolic fractions (HA1-HA9) obtained from the aerial parts of A. vulgaris were evaluated for their antioxidant, antimicrobial and brine shrimp cytotoxic activities. Fraction HA8 showed substantial phenolics content with value of 26.29±1.4μgEQ/mg and DPPH scavenging (82.84±3.01%). Conversely, total flavonoids content of 7.32±0.07μgEQ/mg was determined in HA1 fraction. Fraction HA1 also showed significant cytotoxic effect with the value LD50 of 144.94μg/mL. Fractions HA7 and HA9 depicted maximum total antioxidant activity and ferric ion reduction (96.25±3.29 and AAE/mg and 176.91±8, respectively). All fractions showed encouraging results against bacterial strains Bordetella bronchiseptica and Micrococcus luteus, while HA2 fraction showed the highest percentage inhibition Mucor species with zone of inhibition of 13.25±0.35mm. A total of 7 fractions showed significant antileishmanial activity with survival percentage ranging 0.00 to 19. To sum up, results of the current study indicated that the plant can be further explored for isolation of antileishmanial and antimicrobial compounds, which could be used for drug development.