TY - JOUR T1 - Inhibition of fibrosis with multi-agent therapy in pulmonary fibrosis: Results of a drug library screening JF - bioRxiv DO - 10.1101/2020.06.29.178061 SP - 2020.06.29.178061 AU - Cassandra Batzlaff Braun AU - Megan Girtman AU - Paige Jenson AU - Michael H Bourne AU - JuneMee Chae AU - Theodore Kottom AU - Andrew Limper Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/06/29/2020.06.29.178061.abstract N2 - Aims Successful management of IPF will likely require multi-drug therapy as its pathogenesis is thought to be both driven by both pro-inflammatory and pro-fibrotic pathways. We hypothesized that the available anti-fibrotic agents, pirfenidone and nintedanib, may exhibit synergy in suppressing lung fibroblast extracellular matrix protein generation when administered in combination with other orally active agents.Materials and Methods A fibroblastic cell line (AKR-2B) was stimulated with TGF-β1 and used to screen a library of over 1500 FDA approved drugs. Extracellular matrix protein generation was assessed via fibronectin ELISA assay and maintenance of cell viability confirmed with XTT assay.Results The screening revealed sixty-two drugs from the repurposed drug-screening library that were shown to significantly suppress fibronectin expression and not result in cell death. Specifically drugs within the category of NSAIDs, steroids, azole antifungal agents, and antipyrine were associated with significant suppression of fibronectin on ELISA analysis. Surprisingly, we observed anti-fibrotic activity across a number of the azole antifungal compounds. We next assessed whether combination of azoles would exhibit synergy when combined with current anti-fibrotic therapies in the stimulated fibroblasts. As proof of concept, we demonstrated in vitro synergy between oxiconazole and nintedanib in suppressing fibroblast generation of extracellular matrix fibronectin.Conclusions These results suggest an approach to identify potential combinations of therapy that may improve patient outcomes by reducing cost and potential toxicities during treatment.Competing Interest StatementThe authors have declared no competing interest.(IPF)Idiopathic pulmonary fibrosis(FVC)Forced vital capacity(TGF-β1)Transforming growth factor beta 1(NSAIDs)Non-steroidal anti-inflammatory drugs(ELISA)Enzyme linked immunosorbent assay(NAC)N-Acetyl Cysteine(FDA)Food and Drug Administration(ANOVA)Analysis of variance(FBS)fetal bovine serum(DMSO)Dimethylsulfoxide(PBS)Phosphate buffered saline(RIPA buffer)Radio immunoprecipitation assay buffer(TMB)Tetramethylbenzidine(BAL)Bronchoalveolar lavage ER -