Targeting inflammation through inhibition of COX-2 by substituted 4-thiazolidinone analogues: in-vitro, in-vivo and in-silico studies

Abstract

The prolonged or chronic use of anti-inflammatory agents that are being used clinically is highly unsafe. This flaw or disadvantage has motivated scientists to create new heterocyclic compounds with minimal toxicity and possible anti-inflammatory effects. The design and synthesis of ten 4-thiazolidinone containing pyridine derivatives are reported in the current study article. After conducting an in vitro study, it was discovered that they had anti-inflammatory potential. The compounds were analysed for the COX-2 inhibitory activity. VSA6 found to be most potent candidate, screened further. Also. VSA6 emerged as most potent inhibitor against COX-2 with IC50 of 17.32±1.06µM. Further the antioxidant potential of the candidates was analysed in-vitro data suggested that, synthesized compounds showed the anti-oxidant activity with VSA6, and VSA10 being remarkably potent compounds with IC50 values of 34.39 and 37.32 µM respectively. In RAW 264.7 cells, VSA6 demonstrated dose-dependent suppression of mitochondrial superoxide generation at concentrations of 10 and 20 µM. It decreased intracellular expression of the LPS induced reactive oxygen species; mitochondria induced superoxide, MAPK7 and NF-κβ. The molecule also diminished the expression of extracellular of TNF-α, IL-6, MAPK7, NF-κβ and COX-2 and enhanced the expression of anti-oxidant enzymes i.e. GSH and SOD2. To further validate the COX-2 inhibitory activity of VSA6, gene and protein level expression study was conducted; VSA6 decreased the LPS induced COX-2 expression analysed by flow cytometer, immunofluorescence assay and RT-qPCR. The most efficient compound showed encouraging anti-inflammatory action when tested for in vivo anti-inflammatory potential in carrageenan induced model. To find out how this developed compound interacted with COX-2, an anti-inflammatory target, molecular docking analysis was used. According to the findings, the majority of the compounds could make good candidates for the creation of brand-new anti-inflammatory drugs.