Computational and Preclinical Analysis of 2-(4-Methyl)benzylidene-4,7-dimethyl Indan-1-one (IPX-18): A Novel Arylidene Indanone Small Molecule with Anti-Inflammatory Activity via NF-κB and Nrf2 Signaling

Abstract

Background: The adverse effects of anti-inflammatory drugs urges the search for new anti-inflammatory agents. This study aims at the preclinical analysis of the in-house synthesized small molecule IPX-18. Human whole blood (HWB), peripheral blood mononuclear cells (PBMCs), and neutrophils were used. Rat basophil cells (RBL-2H3) were used to assess degranulation. Binding stability to NF-κB-p50 was predicted using computational docking and molecular dynamic simulations. Essential signaling proteins were evaluated through flow cytometry. Results: IPX-18 inhibited the release of TNF-α with an IC50 value of 298.8 nM and 96.29 nM in the HWB and PBMCs, respectively. The compound depicted an IC50 value of 217.6 nM in the HWB and of 103.7 nM in the PBMCs for IFN-γ inhibition. IL-2 release and IL-8 release were inhibited by IPX-18 in the HWB and PBMCs. The compound controlled the migration of and the elastase in the activated neutrophils. The IC50 value for basophil activation through the FcεRI receptor assay was found to be 91.63 nM. IPX-18 inhibited RBL-2H3-degranulation with an IC50 value of 98.52 nM. The computational docking analysis predicted that IPX-18 would effectively bind NF-κB-p50. NF-κB-phosphorylation in the activated RBL-2H3 cells was decreased, and the levels of nuclear factor erythroid 2-related factor 2 (Nrf2) were increased with IPX-18 treatment. Conclusions: IPX-18 demonstrated efficacy in mediating the effector cells’ inflammatory responses through NF-κB/Nrf2 signaling.

Approaches, Strategies and Procedures for Identifying Anti-Inflammatory Drug Lead Molecules from Natural Products

“…Paprocka et al investigated the anti-inflammatory activity of five natural SMs using LPS- LPS-stimulated PBMC, which showed that all derivatives significantly inhibited the production of pro-inflammatory cytokines, TNF and IFN-γ [ 261 ]. In another study, IPX-18 suppressed the production of IFN-γ, TNF, IL-8 and IL-2, with an IC 50 value of 96.29, 103.7, 122.9 and 105.2 nM by PBMC induced with 50 ng/mL of phorbol-12-myristate-13-acetate (PMA) [ 262 ]. In the IBD clinical setting, TNF increases effector T cells at the site of inflammation, while IFN-γ and IL-2 stimulate immune cells, and IL-8 recruits neutrophils and other immune cells to the site of inflammation [ 263 , 264 , 265 ].…”

“…In the IBD clinical setting, TNF increases effector T cells at the site of inflammation, while IFN-γ and IL-2 stimulate immune cells, and IL-8 recruits neutrophils and other immune cells to the site of inflammation [ 263 , 264 , 265 ]. Hence, the ability of IPX-18 to reduce the production of pro-inflammatory cytokines signifies its anti-inflammatory potency [ 262 ]. Despite being widely used for anti-inflammatory screening, the PBMC assay has shown limitations or phenotypic differences between immune cells and those in the intestinal mucosa [ 237 , 257 ].…”