A Novel Benzenediamine Derivate Rescued Mice from Experimental Sepsis by Attenuating Proinflammatory Mediators via IRAK4

Abstract

We designed and synthesized a novel benzenediamine derivate, FC-99, that was tested for its ability to protect mice from experimental sepsis. Moreover, we sought to determine whether FC-99 could control a bacterial infection and to clarify the mechanism by which FC-99 inhibited LPS-activated macrophages. The effects of FC-99 on inflammation were evaluated in two experimental sepsis models and in cultured macrophages. Microarrays and docking and molecular dynamics simulations were used to determine the target of FC-99. Surface plasmon resonance and molecular detection were performed to confirm the direct interaction of FC-99 with its target. FC-99 protected mice from experimental sepsis. The mice that received FC-99 exhibited a diminished inflammatory response, had a lower local bacterial burden, and experienced a significantly improved survival rate. Genome-wide transcriptional profiling of FC-99-treated macrophages identified IRAK4 as a drug-regulated gene involved in LPS/TLR4 signaling. A computer search and calculations indicated that IRAK4 directly interacted with FC-99. Surface plasmon resonance, IRAK4-regulated signaling pathway analysis, and gene expression profiling of proinflammatory mediators confirmed the direct interaction between FC-99 and IRAK4. FC-99 is a potential therapeutic molecule for sepsis that alleviated experimental sepsis by directly inhibiting IRAK4 activation, which represents a novel target for sepsis therapy.

FC-99 ameliorates sepsis-induced liver dysfunction by modulating monocyte/macrophage differentiation via Let-7a related monocytes apoptosis

“…In the CLP model group, mice displayed 100% mortality at 7 days, whereas, in the 10 mg/kg FC-99 treatment groups, the mortality rate was 32.5%, which indicated an effective improvement of the animal survival ( P < 0.001) (Figure 1A ). Furthermore, the bacterial colony forming unit (CFU) counts in the indicated groups of mice were measured in order to determine whether FC-99 alters the level of bacterial burden in the peritoneal lavage fluid and blood [ 31 ], which were collected 24 hours following the CLP operation. The results indicated that the bacterial CFU counts in the peritoneal lavage fluid and blood of the septic mice were significantly increased ( p < 0.005) compared with the sham-operated mice.…”

Exosomal miR-146a Contributes to the Enhanced Therapeutic Efficacy of Interleukin-1β-Primed Mesenchymal Stem Cells Against Sepsis

“…Western blotting was performed according to previously described methods . Protein concentrations were determined using a Bradford assay (Pierce, Thermo Scientific, Rockford, IL, http://www.thermoscientific.com).…”

TLR4 Signaling Pathway Modulators as Potential Therapeutics in Inflammation and Sepsis

“…In addition to the natural IRAK4 inhibitors mentioned above, the synthetic benzenediamine FC-99 ( Figure 8 ) was proven to bind to IRAK4 both in silico and in vitro (SPR assay) [ 148 ]. FC-99 protected mice in the CLP-induced polymicrobial sepsis model and significantly decreased the serum levels of TNF-α and IL-6.…”