Abstract
Fibrosis is caused by the proliferation of pathogenic myofibroblasts and the deposition of massive amounts of soluble collagen, altering the homeostasis of extracellular matrix (ECM) biogenesis and resulting in tissue scarring. Because a chromone scaffold (CS)-containing small molecule called eupatilin was shown to curb lung fibrosis, a new CS-containing analog, ONG41008, was generated. Plasma exposure was significantly increased. Orally-administered ONG41008 was more potent than pirfenidone at ameliorating fibrosis in a bleomycin-induced lung fibrosis model (BLM). ONG41008 also completely inhibited the trans-differentiation to myofibroblasts of ONGHEPA1, being a primary hepatic stellate cells (HSC) cell line, and of primary diseased human lung fibroblasts (DHLFs) derived from patients with idiopathic pulmonary fibrosis. ONG41008 inhibited the expression of LTBP1 and LAP, dismantling the latent TGFβ complex, likely limiting binding of TGFβ to TGFβ receptors I and II. ONG41008 also markedly inhibited the phosphorylation of SMAD2 and SMAD3, the induction of NADPH oxidase 4 (NOX4) in both cell types, and the production of reactive oxygen species. ONG41008 also completely inhibited the induction of chemokine ligands 2 (Ccl2) and 7 (Ccl7) and induced robust autophagy, suggesting that ONG41008 could curb liver inflammation and fibrosis. STAM mice model was orally administered with 50 mg/kg (mpk) ONG41008 exhibited high nonalcoholic fatty liver disease scores, suggesting that ONG41008, together with anti-diabetic drugs like GLP1 and peroxisome proliferator-activated receptor agonists, could reduce the development of nonalcoholic steatohepatitis (NASH).
Treatment of macrophages with ONG41008 and lipopolysaccharide (LPS) markedly inhibited the expression of TNFα, Interleukin 1β, CHOP, CCL2, CCL7, and CXCL2. ONG41008 also efficiently blocked the endocytosis of CD14, a LPS coreceptor, and retained its expression on the plasma membrane, but had no effect on the cellular distribution of MD-2, TLR4 and Myd88. Taken together, these findings showed that ONG41008 is a bifunctional agent, with both anti-inflammatory and anti-fibrotic activities. This drug may be powerful and safe in the treatment of fibrosis and fibrotic diseases, including NASH, as well as in the treatment of autoimmune, inflammatory, and neuroinflammatory diseases, including rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, and sepsis.
