Abstract
Crataegus azarolus L. (Rosaceae), commonly known as Mediterranean hawthorn, has long been valued in Traditional Medicine for treating cardiovascular and inflammation-related diseases, including diabetes, cancer, and rheumatism. Pharmacological benefits of Crataegus azarolus L. are notably linked to its anti-inflammatory properties. Fupenzic acid, a pentacyclic triterpene isolated from its leaves, holds significant pharmacological potential that remains elusive. This study investigates the unexplored capacity of fupenzic acid as a promising anti-inflammatory agent. Using a multidisciplinary approach that integrates network pharmacology, molecular docking, in vitro assays, and predictive in silico analyses of drug-like properties, ADME, and toxicity, the mechanisms and properties of fupenzic acid have been elucidated. Network pharmacology analysis identified the potential targets for fupenzic acid, with enrichment analyses revealing key processes like inflammatory response, cytokine signaling, innate immune system, and MAPK cascade regulation. Transcription factors such as RELA, SP1, and NFKB1 were predicted to play crucial roles in its therapeutic effects. PPI network analysis underscored NF-κB as a central hub, linking these pathways to its anti-inflammatory effects. In vitro experiments demonstrated that fupenzic acid effectively suppressed inflammatory mediators like NOS-2 and COX-2, through the NF-κB pathway. Molecular docking further confirmed its favorable interaction with NF-κB, reinforcing its mechanism of action. Additionally, in silico ADMET profiling revealed favorable drug-like properties including pharmacokinetics and toxicity profiles, emphasizing its suitability as a drug candidate. This study represents a major step forward in understanding the therapeutic potential of fupenzic acid, establishing it as a distinctive and promising anti-inflammatory agent. The findings identified it as a pharmacological agent for clinical development targeting inflammation-driven diseases and also provide a foundation for future translational research.