The current study presents a series of indomethacin-phenolic phytochemical hybrids (INDO_PP1 to INDO_PP15) using the Steglich esterification reaction to develop potential antioxidant and anti-inflammatory lead drug candidates. After the design of the hybrid candidates (INDO_PP1 to INDO_PP15), various bioinformatics and chemoinformatics tools were employed to predict the potency, toxicity, and drug-ability profiles. The AutoDock 4.2 software was used to assess binding efficacy, GROMACS-2020 for molecular dynamics simulation at 200 nanoseconds; and the binding free energy calculation with the MM/PBSA method to observe and calculate the stability of docking complexes; the SwissADME for physicochemical and pharmacokinetics; and the Molsoft for drug-likeness prediction. Based on the above computational reports, select four hybrids (INDO_PP11, INDO_PP12, INDO_PP14, and INDO_PP15) for synthesis under laboratory conditions and further confirm the structure using different spectral techniques. Cytotoxicity (HEK293, Huh7, and THP1) and antioxidant (DPPH assay) investigation reports suggested that INDO_PP12 was the potential lead, and 20 μM was the optimized therapeutic dose for further anti-inflammatory studies with inflammation (lipopolysaccharide/LPS)-induced THP1 cells. The pro-inflammatory (IL1β, NLRP3), anti-inflammatory (IL10) cytokines and COX2 expression from immunoblotting and real-time PCR suggested that INDO_PP12 showed higher potency than INDO. In summary, the umbelliferone (UMB)-INDO hybrid (INDO_PP12) could be used in place of native INDO for chronic inflammatory and autoimmune disorder management after additional pharmacological investigation.
