The prevalence of multidrug-resistant (MDR) pathogens raises public fears of untreatable infections and represents a huge health risk. There is an urgent need to exploit novel antimicrobial agents. Due to the unique mechanisms, antimicrobial peptides (AMPs) with a low probability to achieve resistance are regarded as potential antibiotic alternatives to address this issue. Herein, we develop a panel of synthetic peptide compounds with novel structures based on the database filters technology (DFT), and the lead peptide LI14 shows potent antibacterial activity against all tested drug-resistant bacteria. LI14 exhibits rapid bactericidal activity and excellent anti-biofilm and -persisters activity, simultaneously showing a low propensity to induce resistance. Moreover, LI14 shows tolerance against pH, temperatures, and pepsin treatment, and no detectable toxicity both in vitro and in vivo. Mechanistic studies revealed that LI14 induces membrane damage by targeting bacterial-specific membrane components and dissipates the proton motive force (PMF), thereby resulting in metabolic perturbations and the accumulation of toxic metabolic products. Furthermore, LI14 sensitizes clinically relevant antibiotics against MDR bacteria. In animal models of infection, LI14 or combined with antibiotics are effective against drug-resistant pathogens. These findings suggest that LI14 is a promising antibiotic candidate to tackle MDR bacterial infections.