<p>Cardioplegic solutions induce cardiac arrest while protecting the myocardium from ischemia reperfusion injury (IRI). Furthermore, they exert effects on organs such as the kidneys and brain, which can impact postoperative outcomes. As our understanding of the IRI-related mechanisms increases, cardioplegic solutions can be improved. Custodiol®-N has been designed on the basis of the Custodiol® cardioplegic solution. This large animal study aims to investigate the protective effects of Custodiol®-N on the myocardium, kidneys and brain. Therefore, German Landrace pigs underwent median sternotomy, cardiopulmonary bypass at 34 °C, 90 min of cardiac arrest and 120 min reperfusion. Animals were randomized for single-shot cardioplegia with either Custodiol®-N (<i>n</i> = 10) or Custodiol® (<i>n</i> = 10). Following reperfusion, tissue sampling and analysis were performed. In the Custodiol®-N group hemoglobin levels and calcium concentrations were more stable after reperfusion. In the myocardial tissue, mitochondrial respiration and integrity as well as the release and activation of hypoxia markers and apoptosis-promoting molecules were comparable. In renal tissue, the tubular structure of proximal tubules was preserved and lower amounts of pro-apoptotic molecules were released in the Custodiol®-N group. In brain tissue, Custodiol®-induced cardioplegia showed higher activation rates of hypoxia-related markers and higher expression of pro-apoptotic cytokines.</p><p>The large animal studies demonstrated that Custodiol®-N-induced cardioplegia provided improved protection against kidney injury and reduced cerebral inflammation while offering comparable myocardial protection compared to Custodiol®-induced cardioplegia.</p>