Background: The present study was undertaken to analyze the phytochemical content and biological activity of CICHORIUM INTYBUS seeds traditionally used in Charsadda, Pakistan against multidrug resistant (MDR) bacterial pathogens. Objectives: This study explored the qualitative and quantitative antibacterial potential of C. INTYBUS. Further qualitative analysis of phytochemical content was performed. Methods: CICHORIUM INTYBUS seed extracts were prepared in aqueous, chloroform, ethanol, and hexane separately. Results: All the extracts of C. INTYBUS seeds were screened for antibacterial activity and phytochemical content. CICHORIUM INTYBUS seed extract showed considerable activity against MDRpathogenic bacteria. In the well diffusion method, aqueous extracts showed a higher zone of inhibition against Pseudomonas aeruginosa (16 mm 0. 7 mm) and Acinetobacter baumannii (13 mm 0. 5 mm), whereas chloroform, ethanol, and hexane extracts showed activity against P. aeruginosa (11 mm 0. 3 mm, 12 mm 0. 5 mm, and 11 mm 0 mm, respectively) as compared to Imipenem, a broad spectrum antibiotic. Minimum inhibitory concentration and minimum bactericidal concentration values for aqueous and ethanol extracts indicate that they were more effective against MDR bacteria. Phytochemical analysis revealed that aqueous and ethanol extracts were rich in alkaloids, carbohydrates, gallotannins, and triterpenoids, whereas chloroform and hexane extracts were more concentrated with phenolics, pseudotannins, saponins, and tannins. CICHORIUM INTYBUS seed extract demonstrated potential activity against MDR human pathogenic bacteria. Conclusions: The undertaken study has for the first time reported the effects of C. INTYBUS seed extracts against MDR bacterial pathogens. Findings of the current study will be helpful for further elucidation of bioactive molecules for therapeutic use against MDR bacterial pathogens.