Modulation of Endothelial Inflammatory Signature by an Imidazo-Pyrazolyl Urea Derivative in a Dynamic In Vitro Model of Hypertension

Hypertension is a multifactorial condition in which inflammation plays a pivotal role. Current preclinical models fail to fully capture the complexity of the underlying mechanisms, limiting therapeutic discovery, while compounds targeting endothelial inflammatory pathways may offer promising alternatives. Methods: An advanced in vitro model of hypertension, consisting of a bioreactor for endothelial cell culture coupled with a peristaltic pump to mimic blood circulation, and a pressure modulator as mechanical stimulus (Live-Pa System) to reproduce hemodynamic stimuli, was employed to investigate the effects of the imidazo-pyrazolyl urea (IPU) 3l, known for its chemotaxis inhibition properties. The effects of 3l were investigated on hypertension-related inflammatory/vasoconstrictor markers, alone or in combination with hypertensive stimuli (ANGII and/or Live-Pa). Results: IPU 3l effectively counteracted ANGII-induced inflammation by significantly reducing NF-κB activation across all experimental conditions (static, dynamic, and Live-Pa) and IL-8 secretion under static and dynamic conditions. Conclusions: IPU 3l exhibits a consistent anti-inflammatory profile, primarily through inhibition of ANGII-induced NF-κB activation across all experimental conditions, with additional context-dependent effects on IL-8 secretion. This study introduces a new strategy for drug discovery by distinguishing biochemical from mechanical stress, providing a clearer framework to interpret condition-specific pharmacological responses. Such insights, difficult to obtain with conventional in vitro or in vivo models, are essential for developing more effective cardiovascular therapies.