2-Butanone 4-glucopyranoside 6'-O-gallate isolated from Memecylon scutellatum exhibits anti-inflammatory activity via modulation of the NF-κB and MAPK signaling pathways in LPS-stimulated RAW 264.7 macrophages.

Inflammation is a major contributor to the development of chronic diseases, including diabetes, rheumatoid arthritis, and cancer. The Memecylon genus has been demonstrated for its diverse biological activities such as antioxidant, antidiabetic, anticancer and antimicrobial effects. Nevertheless, the anti-inflammatory potential of Memecylon scutellatum (Lour.) Hook. & Arn. remains unknown. This study aimed to elucidate the detailed molecular mechanism of 2-butanone 4-glucopyranoside 6'-O-gallate (2-BGG), a bioactive compound isolated from the leaves of Memecylon scutellatum, using lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages as a model. Anti-inflammatory functional outcomes (NO, TNF-α, IL-6) were assessed by Griess and ELISA. The gene expression (RT-qPCR) and protein signaling cascade activation (Western blotting) were employed to define the molecular targets, focusing on the NF-κB and MAPK pathways. Molecular docking and molecular dynamics simulations were performed to investigate the interaction of 2-BGG with IKKβ and MAPK, while physicochemical, pharmacokinetic, and toxicity profiles were predicted using ADMETlab 3.0. The compound was confirmed to be non-cytotoxic at concentrations exhibiting biological activity, where it significantly attenuated the LPS-induced overproduction of NO, TNF-α, and IL-6. This effect was closely associated with the dose-dependent suppression of pro-inflammatory gene products, iNOS and COX-2, at the transcriptional and translational levels. Our investigation revealed that the compound exerts its action by dual modulation of core signaling cascades: it strongly suppressed the phosphorylation and degradation of IκB-α and the activation of NF-κB p65, while simultaneously inhibiting the phosphorylation of p38, ERK1/2, and JNK. In silico analyses supported these findings, suggesting that 2-BGG may bind to the ATP-binding pockets of IKKβ and MAPK in an ATP-competitive manner, and acceptable ADMET properties with low predicted toxicity. These findings support 2-BGG as a potential anti-inflammatory agent that modulates both NF-κB and MAPK signaling pathways. Furthermore, ADMET predictions indicated favorable pharmacokinetic properties and low predicted toxicity. This study provides strong molecular evidence supporting the therapeutic potential of this natural gallate derivative.