Discovery of a Novel Class of Bioactive Molecules Suppressing Pathogenic Fibroblast Activation through Hyou1 Targeting

Fibroblasts play an important role in fibrosis, chronic inflammatory diseases (CIDs) and cancer. Targeting their pathogenicity is crucial, considering the limitations and side effects of existing biologic therapies that focus on inflammatory mediators (TNFα/IL-6). Herein, a multidisciplinary approach utilizing activated primary fibroblasts (PFs) and macrophages (MFs), which are key regulators in inflammation and fibrosis, is presented. The research encompasses medicinal chemistry, molecular phenotyping, mechanism of action studies, RNA-sequencing analysis and ADMET/PK/in vivo evaluation. A novel anti-inflammatory pharmacophore acting through Hypoxia up-regulated protein 1 (Hyou1) was discovered, thus emerging the potential of Hyou1 down-regulation/inhibition in fibroblast-related disorders. The study highlights the need for further investigation into the effects of Hyou1 activity on fibroblasts while providing the first reported Hyou1 small molecule inhibitor leads for drug discovery. To investigate the deregulated genes on activated fibroblasts upon compound 33 treatment, we performed 3prime UTR sequencing comparing compound treated hTNFtg synovial fibroblasts versus the vehicle (dmso) treated controls