A novel cathepsin L inhibitor prevents the progression of idiopathic pulmonary fibrosis

[Display omitted] •Compound 3 showed 1.5-fold more potent activity than that of control against Cat L.•Compound 3 presented new interaction modes with cathepsin by its B-ring moiety.•MD simulation showed that 3 and Cat L kept stable interaction in the binding site.•The anti-fibrotic ability of 3 was confirmed by in vitro and in vivo study.•It is the first report that asperphenamate analog showed the anti-fibrotic ability. In previous work, the target of asperphenamate as a natural product was successfully determined as cathepsin by the natural product consensus pharmacophore strategy. In order to develop accurate SAR (structure-activity relationship) of asperphenamate-type cathepsin inhibitor, we chose several novel analogs with heterocyclic moiety to perform further study. The molecular simulation showed that 4-pyridyl derivative 3 with the greatest cathepsin inhibitory activity presented new interaction modes with protein utilizing its B-ring moiety. And then molecular dynamics (MD) simulation further revealed that 3 and cathepsin kept stable interaction in the binding site, which validated the molecular docking results. In view that cathepsins play an important role in fibrosis and some cathepsin inhibitors display the therapeutic ability for fibrosis, we investigated the anti-fibrotic effect of 3in vitro and in vivo. The results indicated that 3 displayed the strongest inhibitory effect on the formation of α-SMA and collagen I as the protein markers of fibrosis among all tested derivatives. Further in vivo assay confirmed that 3 indeed showed significant inhibitory ability against pulmonary fibrosis by the method of H&E and Masson staining as well as immunohistochemical staining for characteristic α-SMA proteins.