Anti-inflammatory activity of the piperazino-enaminone JODI-19 in vitro and in vivo

Abstract

Abstract Data to characterize novel anti-inflammatory agent enaminone E121 have shown a marked decrease in the release of pro-inflammatory cytokines in macrophages stimulated with lipopolysaccharide (LPS) compared to the control (DMSO) group. Literature has described that when an N-alkylated piperazine motif is added to the terminal end of the lead compound, it is able to increase the CCR2 antagonistic activity. We hypothesized that incorporating the N-alkylated piperazine motif into the existing E-121 enaminone pharmacophore could lead to a ligand with dual functionality as a cytokine inhibitor and chemokine receptor antagonist. The resultant series (JODI) were studied in vitro, and data suggest they can suppress TNF-alpha and IL-6 in a dose-dependent manner, superior to that of dexamethasone. Additionally, macrophages stimulated with chemokine MCP-1 and treated with JODI compounds, (but not E121), are able to block CCR2 signaling as evidenced by decreased total ERK1/2. To determine if JODI-19 has activity in vivo, it was utilized in a mouse model of hemarthrosis to assess decreased inflammation as evidenced by local cytokine suppression. In a pilot study, FIX−/−mice were subjected to hemarthrosis induction with or without JODI administration. At different timepoints from four hours to four weeks after hemarthrosis, synovial fluid was collected by lavage for multiple cytokine analyses. Administration of JODI-19 significantly decreased TNFα, IL-1α, MCP-1 and MIP-1α levels in the synovial fluid. As such, JODI-19 may be a novel approach to better manage hemophilia arthropathy and its associated bone damage.