title: EW-7197 inhibits hepatic, renal, and pulmonary fibrosis by blocking TGF-β/Smad and ROS signaling.
abstract: Fibrosis is an inherent response to chronic damage upon immense apoptosis or necrosis. Transforming growth factor-beta1 (TGF-β1) signaling plays a key role in the fibrotic response to chronic liver injury. To develop anti-fibrotic therapeutics, we synthesized a novel small-molecule inhibitor of the TGF-β type I receptor kinase (ALK5), EW-7197, and evaluated its therapeutic potential in carbon tetrachloride (CCl4) mouse, bile duct ligation (BDL) rat, bleomycin (BLM) mouse, and unilateral ureteral obstruction (UUO) mouse models. Western blot, immunofluorescence, siRNA, and ChIP analysis were carried out to characterize EW-7197 as a TGF-β/Smad signaling inhibitor in LX-2, Hepa1c1c7, NRK52E, and MRC5 cells. In vivo anti-fibrotic activities of EW-7197 were examined by microarray, immunohistochemistry, western blotting, and a survival study in the animal models. EW-7197 decreased the expression of collagen, α-smooth muscle actin (α-SMA), fibronectin, 4-hydroxy-2, 3-nonenal, and integrins in the livers of CCl4 mice and BDL rats, in the lungs of BLM mice, and in the kidneys of UUO mice. Furthermore, EW-7197 extended the lifespan of CCl4 mice, BDL rats, and BLM mice. EW-7197 blocked the TGF-β1-stimulated production of reactive oxygen species (ROS), collagen, and α-SMA in LX-2 cells and hepatic stellate cells (HSCs) isolated from mice. Moreover, EW-7197 attenuated TGF-β- and ROS-induced HSCs activation to myofibroblasts as well as extracellular matrix accumulation. The mechanism of EW-7197 appeared to be blockade of both TGF-β1/Smad2/3 and ROS signaling to exert an anti-fibrotic activity. This study shows that EW-7197 has a strong potential as an anti-fibrosis therapeutic agent via inhibition of TGF-β-/Smad2/3 and ROS signaling.
authors.affiliation: College of Pharmacy, Ewha Womans University, Seodaemun-gu, Seoul, 120-750, South Korea.
authors.forename: Sang-A
authors.initials: SA
authors.lastname: Park
authors.forename: Min-Jin
authors.initials: MJ
authors.lastname: Kim
authors.forename: So-Yeon
authors.initials: SY
authors.lastname: Park
authors.forename: Jung-Shin
authors.initials: JS
authors.lastname: Kim
authors.forename: Seon-Joo
authors.initials: SJ
authors.lastname: Lee
authors.forename: Hyun Ae
authors.initials: HA
authors.lastname: Woo
authors.forename: Dae-Kee
authors.initials: DK
authors.lastname: Kim
authors.forename: Jeong-Seok
authors.initials: JS
authors.lastname: Nam
authors.forename: Yhun Yhong
authors.initials: YY
authors.lastname: Sheen
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology*; D000818:Animals; D001761:Bleomycin; D015153:Blotting, Western; D002251:Carbon Tetrachloride; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers / Q000235:genetics; D005355:Fibrosis / Q000517:prevention & control*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008168:Lung / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051785:Smad Proteins / Q000378:metabolism; D014230:Triazoles / Q000494:pharmacology*
chemical_list: D000814:Aniline Compounds; D017931:DNA Primers; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D018125:Receptors, Transforming Growth Factor beta; D051785:Smad Proteins; D014230:Triazoles; D001761:Bleomycin; C000590371:vactosertib; D002251:Carbon Tetrachloride; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human; C000625842:Tgfbr1 protein, mouse; C000625843:Tgfbr1 protein, rat