NADPH oxidase 2 inhibitors CPP11G and CPP11H attenuate endothelial cell inflammation & vessel dysfunction and restore mouse hind-limb flow

• Published in: Redox biology Vol. 22; p. 101143
• Main Authors: Li, Y, Cifuentes-Pagano, E, DeVallance, E R, de Jesus, D S, Sahoo, S, Meijles, D N, Koes, D, Camacho, C J, Ross, M, St Croix, C, Pagano, P J
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier 01.04.2019
• Subjects: Animals; Biomarkers; Cell Adhesion; Cell Line; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Hindlimb - blood supply; Hindlimb - metabolism; Humans; Mice; Models, Molecular; Molecular Conformation; Monocytes - drug effects; Monocytes - metabolism; NADPH Oxidase 2 - antagonists & inhibitors; NADPH Oxidase 2 - chemistry; NADPH Oxidase 2 - metabolism; NF-kappa B - metabolism; Protein Binding; Protein Interaction Domains and Motifs; Reactive Oxygen Species - metabolism; Regional Blood Flow - drug effects; Research Paper; Signal Transduction; Structure-Activity Relationship; Transcription Factor AP-1 - metabolism; Vasculitis - drug therapy; Vasculitis - etiology; Vasculitis - metabolism; Vasculitis - pathology; Endothelial dysfunction; Reactive oxygen species; Vascular inflammation; Small-molecule inhibitor; NADPH oxidase
• ISSN: 2213-2317; 2213-2317
• DOI: 10.1016/j.redox.2019.101143

First described as essential to the phagocytic activity of leukocytes, Nox2-derived ROS have emerged as mediators of a range of cellular and tissue responses across species from salubrious to deleterious consequences. Knowledge of their role in inflammation is limited, however. We postulated that TNFα-induced endothelial reactive oxygen species (ROS) generation and pro-inflammatory signaling would be ameliorated by targeting Nox2. Herein, we in silico-modelled two first-in-class Nox2 inhibitors developed in our laboratory, explored their cellular mechanism of action and tested their efficacy in in vitro and mouse in vivo models of inflammation. Our data show that these inhibitors (CPP11G and CPP11H) disrupted canonical Nox2 organizing factor, p47 , translocation to Nox2 in the plasma membrane; and abolished ROS production, markedly attenuated stress-responsive MAPK signaling and downstream AP-1 and NFκB nuclear translocation in human cells. Consequently, cell adhesion molecule expression and monocyte adherence were significantly inhibited by both inhibitors. In vivo, TNFα-induced ROS and inflammation were ameliorated by targeted Nox2 inhibition, which, in turn, improved hind-limb blood flow. These studies identify a proximal role for Nox2 in propagated inflammatory signaling and support therapeutic value of Nox2 inhibitors in inflammatory disease.