Barleria prionitis is a famous perennial plant commonly known as porcupine flower or Vajradanti. It is a shrub with yellow flowers, inhabit most parts of India. Various parts of the plant such as leaves, roots, aerial parts, flowers, and stems are used in the traditional system of medicine. The objective of the study was undertaken to investigate the antimicrobial and wound healing activity of Barleria prionitis leaves extract . The leaves of Barleria prionitis were collected, processed & standardized as per official methods. The extracts of Barleria prionitis leaves were extracted and studied to detect the chemical compounds present by defattying with Petroleum-ether60-800C then successive extraction was done by Ethylacetate & Methanol. Qualitative phytochemical screening revealed the presence of carbohydrates, proteins, flavonoids, tannins, alkaloids & steroids. Total polyphenol content & total flavonoid content was determined using UV-Visible spectrophotometer. DPPH scavenging assay were performed to evaluate the antioxidant capacity of all extracts & they showed maximum antioxidant activity. Then these extracts used to evaluate acute oral toxicity, antioxidant, antimicrobial and wound healing activity. The evaluation of in-vitro antimicrobial and in vivo wound healing activity in rodents was done using various experimental models. Excision model methods were performed to evaluate the wound healing activity by using 1% as well as 2% methanolic extracts ointments and 1% as well as 2% ethyl acetate extracts ointments , in both the extract ointments the rats showed the decreased epithelial size of wound on post wounding days for both 1% as well as 2% methanolic extract and 1% as well as 2% ethyl acetate leaves extract compared to standard group. Along with this the antimicrobial activity was done by using both extracts. In the evaluation of antimicrobial activity , it was found that the Barleria prionitis leaves extracts exhibit significant Antibacterial activity on microorganisms like Bacillus substillus, Staphylococcus aureus & E.coli. The present study shows that Barleria prionitis leaves extract have better wound healing and antimicrobial activity.