Background: Urinary tract infections refer to the presence of harmful microorganisms in either the urethra or bladder or the ureter and pelvis of the kidney. Bacteria that live in environments with low iron concentrations create siderophores. Objectives: This research aims to determine antimicrobial activity of siderophores on multidrug resistant bacteria, Candida albicans (C. albicans) and anticancer activity of siderophore on cancer cell lines. Methods: This study was conducted using a cross-sectional design for 100 urine samples obtained from Iraqi patients suspected of having UTI. Who visited the Baghdad teaching hospital. 45 of the samples were multidrug resistant bacteria cultured on blood agar, and 20 of the samples were Candida albicans cultured on Sabouraud dextrose agar and incubated at 37 °C for 24 hours. The diagnosis was documented using the Vitek 2 system. Siderophores were extracted from a UPEC standard strain by using ethylacetate, purified by Sephadex LH-20 gel and identification by HPLC. Siderophore was detected was done using Chrome azurol S in M9 minimal salts medium. Results: The siderophores demonstrated significant antibacterial effects against multidrug resistant bacteria on Gram positive and negative but its effect is higher in Gram positive than Gram negative. Furthermore, siderophores don't demonstrate any effect on C. albicans. The cytotoxic effects of these siderophores were tested on breast cancer cell lines to assess their potential as anticancer agents. Conclusion: Siderophores extracted and partially purified from UPEC have both anticancer activity against breast cancer and antimicrobial activity against challenging infections like those caused by multidrug resistant bacteria.