Antibiotic resistance has become one of the most urgently important problems facing healthcare providers. A novel series of dipicolylamine-containing carbazole amphiphiles with strong Zn2+chelating ability were synthesized, biomimicking cationic antimicrobial peptides. Effective broad-spectrum 16combined with 12.5 μg/mL Zn2+was identified as the most promising antimicrobial candidate. 16combined with 12.5 μg/mL Zn2+exhibited excellent antimicrobial activity against both Gram-positive and Gram-negative bacteria (MICs = 0.78–3.125 μg/mL), weak hemolytic activity, and low cytotoxicity. Time-kill kinetics and mechanism studies revealed 16combined with 12.5 μg/mL Zn2+had rapid bacterial killing properties, as evidenced by disruption of the integrity of bacterial cell membranes, effectively preventing bacterial resistance development. Importantly, 16combined with 12.5 μg/mL Zn2+showed excellent in vivoefficacy in a murine keratitis model caused by Staphylococcus aureusATCC29213 or Pseudomonas aeruginosaATCC9027. Therefore, 16combined with 12.5 μg/mL Zn2+could be a promising candidate for treating bacterial infections.