Bacterial quorum sensing (QS) and bacterial extracellular polymeric substances (EPS) are the major challenges in treating biofilm chronic infection. Herein, we proposed targeting intervention phase separation of EPS by nano-antibiotics (Ir(ppy)2dmb-PAZ-HA) as an effective strategy for combating bacterial biofilm infection, in which Iridium complex (Ir(ppy)2dmb-CHO) and antibiotic (Pazufloxacin, PAZ) for synergistic antibacterial, and hyaluronic acid (HA) was used to enhance Ir(ppy)2dmb-PAZ release on-demand in biofilm microenvironment. In vitro results showed that Ir(ppy)2dmb-PAZ-HA holds excellent antibacterial activity and antibiofilm capacity. Importantly, Ir(ppy)2dmb-PAZ-HA can be target to regulate phase separation of EPS, and then interfere with biofilm formation, which is a competitive mechanism between Ir(ppy)2dmb-PAZ-HA and Methicillin-resistant Staphylococcus aureus (MRSA) itself. In addition, in vivo results confirmed that Ir(ppy)2dmb-PAZ-HA was able to accelerate biofilm chronic wound healing, and effectively alleviate bacterial acute lung infection. Accordingly, this study provides a new antibiofilm target, and provides a novel idea for the design of small molecular metal antibacterial drugs and antibiotic reuse.