The cardioprotective potential of Robinin (Rob) is hindered by its poor stability, low bioavailability, and rapid degradation. To address these challenges, we developed an Atrial Natriuretic Peptide (ANP)-conjugated, Rob-loaded poly(lactic-co-glycolic acid)-polyethylene glycol (PLGA-PEG) nanoparticle system (NpR) for prolonged and targeted delivery to cardiomyocytes. We hypothesized that ANP-tagged PLGA-PEG nanoparticle would enhance Rob’s therapeutic efficacy by providing sustained release, extended bioactivity, and reduced cytotoxicity. NpR was synthesized via a double emulsion technique and characterized using Fourier Transform Infrared Spectroscopy (FTIR), Energy Dispersive X-ray (EDX) analysis, Dynamic Light Scattering (DLS), and Scanning Electron Microscopy (SEM). Particle size, encapsulation efficiency, peptide and drug loading were measured. In vitro drug release kinetics, cellular uptake, and cytotoxicity in H9c2 cardiomyocytes were assessed. Apoptosis and pro-survival pathway analyses were performed. Characterization revealed that NpR had a particle size of 166.6 ± 2.45 nm, a polydispersity index of 0.225 ± 0.089, and a zeta potential of − 19.00 ± 1.11 mV, with spherical morphology confirmed by SEM. Encapsulation and loading efficiencies of Rob were 75.54 ± 3.11% and 10 ± 0.988% respectively, with an 81 ± 3.98% cumulative Rob release. NpR exhibited a four-fold increase in H9c2 cell uptake, mitigating ISO-induced damage over 24, 48, and 72 h by reducing ROS, apoptosis, and preserving cellular structure. Gene expression analysis showed that NpR reversed ISO-induced alterations in pro- and anti-apoptotic gene levels, while western blotting confirmed restoration of AKT/GSK3β and ERK1/2 signaling pathways. These findings demonstrate that NpR confers potent cardioprotective effects through targeted delivery, sustained release, and enhanced cellular uptake, highlighting its potential as a novel therapeutic strategy for myocardial protection.