Synthesis, Characterization, and Anti‐Inflammatory Profiling of Ibuprofen–Chalcone Conjugate With RP‐HPLC Method Development for Quantification

Inflammation is a major pathological condition associated with various chronic diseases, including arthritis, cardiovascular disorders, and neurodegenerative diseases. Non‐steroidal anti‐inflammatory drugs, like ibuprofen, are widely used to manage inflammation but are often associated with adverse‐effects, including gastrointestinal toxicity. Herein, a novel chloro‐substituted ibuprofen–chalcone conjugate (Compound 6 ) was synthesized to combine anti‐inflammatory potency of ibuprofen with multitargeted activity of chalcones, potentially reducing side‐effects and enhancing therapeutic efficacy. The conjugate's structure was confirmed through various spectroscopic techniques. Conjugate was evaluated for in‐vitro cyclooxygenases‐1/‐2 inhibitory activity along with in‐vivo anti‐inflammatory, analgesic, and ulcerogenic properties at doses of 0.12 and 0.24 mmol/kg. A validated reverse phase‐HPLC method, adhering to International Council on Harmonization Q2 (R1) guidelines, was developed for its quantification, covering parameters like accuracy, precision, linearity, limit of detection and quantitation, and solution stability. In‐vitro cyclooxygenase inhibition assay depicted that Compound 6 exhibited enhanced cyclooxygenase‐1 inhibition than ibuprofen while retaining a comparable selectivity profile. In‐vivo anti‐inflammatory activity using carrageenan‐induced paw edema model showed that conjugate significantly reduced inflammation compared to ibuprofen, with highest dose displaying superior efficacy. Analgesic activity assessed by acetic acid‐induced writhing and hot plate tests revealed dose‐dependent enhancement, with 0.24 mmol/kg dose outperforming ibuprofen. Importantly, ulcerogenic studies demonstrated markedly lower ulcer scores across all prodrug doses. Even at higher dose, the conjugate maintained a better safety profile than ibuprofen, reinforcing its potential for clinical application in managing inflammation with reduced gastrointestinal side‐effects. Further, free drug release after hydrolysis was assessed using the proposed chromatographic method, which confirmed that hydrolysis releases the parent drugs primarily at intestinal pH, minimizing gastric exposure. Overall, ibuprofen–chalcone conjugate ( 6 ) shows promise as a multi‐functional hybrid molecule with improved anti‐inflammatory and analgesic efficacy and reduced gastrointestinal toxicity, making it a strong candidate for further development as a safer alternative against inflammation.