Tetroxoprim is primarily utilized as an antibacterial agent, particularly in the treatment of urinary tract infections (UTIs). It functions by inhibiting dihydrofolate reductase, an enzyme crucial for bacterial DNA synthesis, thereby preventing the growth and reproduction of bacteria. This makes it effective against a range of Gram-positive and Gram-negative bacteria. Tetroxoprim is often combined with sulfonamides to enhance its efficacy, as the combination targets different stages of bacterial folate metabolism, leading to a synergistic antibacterial effect. Its application is particularly valuable in cases where other antibiotics may be less effective or where resistance is a concern.