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 TGF complex, likely limiting binding of TGF to TGF 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 TNF, Interleukin 1, CHOP, CCL2, CCL7, and CXCL2.

Uncontrolled proliferative diseases, such as fibrosis or cancer, can be fatal. We previously found that a compound containing the chromone scaffold (CS), ONG41008, had potent antifibrogenic effects associated with EMT or cell-cycle control resembling tumorigenesis. We investigated the effects of ONG41008 on tumor cells and compared these effects with those in pathogenic myofibroblasts. Stimulation of A549 (lung carcinoma epithelial cells) or PANC1 (pancreatic ductal carcinoma cells) with ONG41008 resulted in robust cellular senescence, indicating that dysregulated cell proliferation is common to fibrotic cells and tumor cells. The senescence was followed by multinucleation, a manifestation of mitotic slippage. There was significant upregulation of expression and rapid nuclear translocation of p-TP53 and p16 in the treated cancer cells, which thereafter died after 72 h confirmed by 6 day live imaging. ONG41008 exhibited a comparable senogenic potential to that of dasatinib. Interestingly, ONG41008 was only able to activate caspase-3, 7 in comparison with quercetin and fisetin, also containing CS in PANC1. ONG41008 did not seem to be essentially toxic to normal human lung fibroblasts or primary prostate epithelial cells, suggesting ONG41008 can distinguish the intracellular microenvironment between normal cells and aged or diseased cells. This effect might occur as a result of the increased NAD/NADH ratio, because ONG41008 restored this important metabolic ratio in cancer cells. Taken together, this is the first study to demonstrate that a small molecule can arrest uncontrolled proliferation during fibrogenesis or tumorigenesis via both senogenic and senolytic potential. ONG41008 could be a potential drug for a broad range of fibrotic or tumorigenic diseases.

In brain ischemia, oxidative stress induces neuronal apoptosis, which is mediated by increased activity of the voltage-gated K+ channel Kv2.1 and results in an efflux of intracellular K+. The molecular mechanisms underlying the regulation of Kv2.1 and its activity during brain ischemia are not yet fully understood. Here this study provides evidence that oxidant-induced apoptosis resulting from brain ischemia promotes rapid tyrosine phosphorylation of Kv2.1. When the tyrosine phosphorylation sites Y124, Y686, and Y810 on the Kv2.1 channel are mutated to non-phosphorylatable residues, PARP-1 cleavage levels decrease, indicating suppression of neuronal cell death. The tyrosine residue Y810 on Kv2.1 was a major phosphorylation site. In fact, cells mutated Y810 were more viable in our study than were wild-type cells, suggesting an important role for this site during ischemic neuronal injury. In an animal model, tyrosine phosphorylation of Kv2.1 increased after ischemic brain injury, with an observable sustained increase for at least 2 h after reperfusion. These results demonstrate that tyrosine phosphorylation of the Kv2.1 channel in the brain may play a critical role in regulating neuronal ischemia and is therefore a potential therapeutic target in patients with brain ischemia.