Design, synthesis, and biological evaluation of ACF-03: a novel anti-fibrotic and antioxidant agent targeting ROS-TGF-β1-HIF-1α axis

• Published in: Applied biological chemistry Vol. 68; no. 1; p. 44
• Main Authors: Abeysiriwardhana, Hiruni Nilshi Indeevarie, Choi, Jin-Hyuk, Anandappa, Joshua Miguel, Malla, Ayusha, Cho, Moonjae
• Format: Journal Article
• Language: English
• Published: Singapore Springer Nature Singapore 01.12.2025; Springer Nature B.V; SpringerOpen; 한국응용생명화학회
• Subjects: ACF-03; Antifibrotic therapy; antioxidant activity; Antioxidants; Applied Microbiology; Biocompatibility; biological assessment; Biological Techniques; Bioorganic Chemistry; blood proteins; Blood-brain barrier; Cell culture; Chemistry; Chemistry and Materials Science; Clinical trials; Collagen (type I); compliance; computer simulation; Connective tissue growth factor; Design analysis; disease progression; Drug development; Drug interaction; Extracellular matrix; Fibroblasts; Fibrosis; Flavonoids; Glycerol; Guaiacol; Hydrogenation; Hypoxia; Hypoxia-inducible factor 1a; In vivo methods and tests; IPF; Lung diseases; moieties; Oxidative stress; permeability; Pharmacokinetics; Pharmacology; Pulmonary fibrosis; renal clearance; ROS-TGF-β1-HIF-1α axis; Synthesis; therapeutics; Toxicity; Transforming growth factor-b1; Vascular endothelial growth factor; 농학; United States > US; Oxidative stress; Antifibrotic therapy; ROS-TGF-β1-HIF-1α axis; IPF; ACF-03
• ISSN: 2468-0842; 2468-0834; 2468-0842
• DOI: 10.1186/s13765-025-01007-8

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease characterized by excessive extracellular matrix deposition, oxidative stress, and dysregulated TGF-β1 and HIF-1α signalling. Current antifibrotic therapies, such as Nintedanib and Pirfenidone, slow disease progression but fail to reverse established fibrosis, necessitating the development of multi-targeted therapeutic agents. This study aimed to design, synthesize, and evaluate ACF-03, a flavonoid-based antifibrotic compound, targeting the ROS-TGF-β1-HIF-1α axis to mitigate oxidative stress-induced fibrotic remodelling. ACF-03 was synthesized by modifying a 4′,6,7-trimethoxyisoflavone (TMF) scaffold with a guaiacol moiety to enhance its antioxidant and antifibrotic properties. The compound’s effects on ROS levels, HIF-1α expression, and fibrosis-related markers were evaluated in TGF-β1-stimulated A549 and HLF cells, with Nintedanib as a positive control. In silico ADMET profiling was performed using SwissADME and ADMET Lab 2.0.ACF-03 exhibited potent antioxidant activity, significantly reducing intracellular ROS levels, downregulating HIF-1α, and suppressing VEGF and CTGF expression. It attenuated fibrotic markers (FN1, COL1A1, and COL3A1, α-SMA) in both cell models, with efficacy comparable to Nintedanib. Pharmacokinetic analysis confirmed compliance with Lipinski’s Rule of Five, efficient permeability, renal clearance potential, and moderate plasma protein binding (PPB < 90%), suggesting a wide therapeutic index and reduced drug-drug interactions. Additionally, CBrain/CBlood < 1 indicated an inability to cross the blood–brain barrier (BBB), minimizing CNS-related toxicity. ACF-03 exhibits potent antifibrotic, antioxidant, and HIF-1α inhibitory via promoting proteosome degradation, making it a promising candidate for IPF treatment. Its multi-targeted mechanism, favourable pharmacokinetics, and high therapeutic index justify further in vivo and preclinical investigations to confirm its clinical translatability.