The discovery of bioactive natural compounds from microbes holds promise for regenerative medicine. In this study, we identified and characterized a steroid-like compound, 3,12-dioxochola-4,6-dien-24-oic acid (DOCDA), from a crude extract of Rhodococcus sp. DOCDA significantly promoted wound healing by enhancing HaCaT cell invasion and migration. It upregulated key growth factors (EGF, VEGF-A, IGF, TGF-β, and HGF), indicating the activation of regenerative signaling. Additionally, DOCDA increased the expression of genes related to focal adhesion and cytoskeletal regulation (ITGB1, ITGA4, FAK, SRC, RHOA, CDC42, RAC1, and paxillin), supporting enhanced cellular motility and remodeling. Notably, DOCDA promoted stem-like properties in HaCaT cells, as shown by increased spheroid formation and elevated levels of the stemness markers ALDH1 and CD44. Target prediction and molecular docking identified the glucocorticoid receptor (GR) as the primary target of DOCDA, with a docking score of −7.7 kcal/mol. Network and pathway enrichment analysis revealed that GR-linked pathways were significantly associated with wound healing, including steroid hormone signaling, inflammation, immune responses, and cell migration. In vivo, the topical application of DOCDA led to over 70% wound closure in mice by day 5. These findings suggest that DOCDA is a steroid-like compound that accelerates wound healing and may serve as a potential agent in regenerative therapy.

Objective: This study aimed to isolate and identify bioactive secondary metabolites, including a new phenazine derivative, from a marine-derived Streptomyces sp. CNQ-031 belonging to the MAR4 clade, and to evaluate their antibacterial activity. Methods: The marine-derived Streptomyces sp. was cultivated and extracted using ethyl acetate. The crude extracts were subjected to chromatographic separation to afford a new natural product, pontophenazine ( 1 ), along with known compounds, 1- methoxyphenazine ( 2 ), 5,7-dihydroxy-2-(1-methylpropyl)-4H-chromen-4-one ( 3 ), and 5,7-dihydroxy-2-isopropyl-4H-chromen-4-one ( 4 ). The chemical structure of 1 was elucidated through analysis of nuclear magnetic resonance (NMR) and mass spectrometry (MS) data, while the chemical structures of 2 − 4 were identified by comparing their NMR data with those of previously reported compounds. Antibacterial activity was assessed using the broth microdilution method. Results: Four compounds were isolated, including a new phenazine derivative, pontophenazine ( 1 ) from a marine-derived Streptomyces sp. CNQ-031. This compound exhibited antibacterial activity against B. subtilis KCTC 1021, with a minimum inhibitory concentration (MIC) value of 64 μg/mL, showed significant biofilm inhibition towards A. tumefaciens SND195, and P. aeruginosa SNC165 (35.4 ± 1.1%, 19.7 ± 0.8%, respectively), and displayed weak inhibitoryactivity against M. luteus and S. aureus in the quorum-sensing assay. Conclusion: The discovery of pontophenazine ( 1 ) expands the structural diversity of marine-derived phenazine derivatives and highlights the MAR4 Streptomyces clade as a valuable source of new antibacterial compounds.