The ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are major causes of hospital-acquired infections worldwide, often exhibiting multidrug resistance. This resistance is a significant challenge in clinical practice and a top global public health threat, driven by excessive antibiotic use, improper prescriptions, and substandard drugs. Understanding resistance mechanisms is crucial for developing new antimicrobial agents or alternative treatments. This review summarizes the known antimicrobial resistance mechanisms of ESKAPE pathogens. Resistance mechanisms include drug inactivation by enzymes like β-lactamases, modification of drug targets such as penicillin-binding proteins, reduced intracellular drug accumulation through porin loss or efflux pumps, and biofilm formation. β-lactamases, including ESBLs and carbapenemases, are prevalent and contribute to resistance against various antibiotics. Efflux pumps, such as those in the RND family, help bacteria expel drugs, while biofilms provide a protective environment that reduces drug effectiveness. Enterococcus faecium often exhibits resistance through low-affinity penicillin-binding proteins and vancomycin resistance. Staphylococcus aureus resistance is associated with β-lactamase production and the emergence of MRSA and VISA/VRSA. Klebsiella pneumoniae and Acinetobacter baumannii show resistance through carbapenemases and other enzymes. Pseudomonas aeruginosa resistance involves AmpC production and porin changes. Enterobacter species exhibit a range of resistance mechanisms, including ESBLs and carbapenemases. These pathogens pose significant challenges due to their resistance mechanisms, which include enzymatic inactivation, target modification, altered permeability, and biofilm formation. Antimicrobial resistance in ESKAPE pathogens is a major threat to global public health and requires new therapeutic strategies, such as antivirulence approaches, bacteriophage therapy, and biofilm inhibitors. Current research focuses on developing new antibiotics and alternative treatments to combat these resistant pathogens.The ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are major causes of hospital-acquired infections worldwide, often exhibiting multidrug resistance. This resistance is a significant challenge in clinical practice and a top global public health threat, driven by excessive antibiotic use, improper prescriptions, and substandard drugs. Understanding resistance mechanisms is crucial for developing new antimicrobial agents or alternative treatments. This review summarizes the known antimicrobial resistance mechanisms of ESKAPE pathogens. Resistance mechanisms include drug inactivation by enzymes like β-lactamases, modification of drug targets such as penicillin-binding proteins, reduced intracellular drug accumulation through porin loss or efflux pumps, and biofilm formation. β-lactamases, including ESBLs and carbapenemases, are prevalent and contribute to resistance against various antibiotics. Efflux pumps, such as those in the RND family, help bacteria expel drugs, while biofilms provide a protective environment that reduces drug effectiveness. Enterococcus faecium often exhibits resistance through low-affinity penicillin-binding proteins and vancomycin resistance. Staphylococcus aureus resistance is associated with β-lactamase production and the emergence of MRSA and VISA/VRSA. Klebsiella pneumoniae and Acinetobacter baumannii show resistance through carbapenemases and other enzymes. Pseudomonas aeruginosa resistance involves AmpC production and porin changes. Enterobacter species exhibit a range of resistance mechanisms, including ESBLs and carbapenemases. These pathogens pose significant challenges due to their resistance mechanisms, which include enzymatic inactivation, target modification, altered permeability, and biofilm formation. Antimicrobial resistance in ESKAPE pathogens is a major threat to global public health and requires new therapeutic strategies, such as antivirulence approaches, bacteriophage therapy, and biofilm inhibitors. Current research focuses on developing new antibiotics and alternative treatments to combat these resistant pathogens.