Infections caused by methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) continue to endanger public health. Here, we report the synthesis of neolignan isomagnolone (<b>I</b>) and its isomer <b>II</b>, and the preparation of a series of novel neolignan-antimicrobial peptide (AMP) mimic conjugates. Notably, conjugates <b>III5</b> and <b>III15</b> exhibit potent anti-MRSA activity <i>in vitro</i> and <i>in vivo</i>, comparable to that of vancomycin, a current effective treatment for MRSA. Moreover, <b>III5</b> and <b>III15</b> display not only fast-killing kinetics and low resistance frequency but also low toxicity as well as effects on bacterial biofilms. Mechanism studies reveal that <b>III5</b> and <b>III15</b> exhibit rapid bactericidal effects through binding to the phosphatidylglycerol (PG) and cardiolipin (CL) of the bacterial membrane, thereby disrupting the cell membranes and allowing increased reactive oxygen species (ROS) as well as protein and DNA leakage. The results indicate that these neolignan-AMP mimic conjugates could be promising antimicrobial candidates for combating MRSA infections.