LCC-09, a Novel Salicylanilide Derivative, Exerts Anti-Inflammatory Effect in Vascular Endothelial Cells

Abstract

Ramcharan Singh Angom,1,* Jian Zhu,1,2,* Alexander TH Wu,3 Maryam Rachmawati Sumitra,4 Victoria Pham,1 Shamit Dutta,1 Enfeng Wang,1 Vijay Sagar Madamsetty,1 Gabriel D Perez-Cordero,1 Hsu-Shan Huang,4 Debabrata Mukhopadhyay,1 Ying Wang5,6 1Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Jacksonville, FL, 32224, USA; 2Department of Cardiology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, 233004, People’s Republic of China; 3The Ph.D. Program for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan; 4Graduate Institute for Cancer Biology & Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei, 110, Taiwan; 5Department of Cardiovascular Medicine, College of Medicine and Science, Mayo Clinic, Rochester, MN, USA; 6Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Rochester, MN, 55905, USA*These authors contributed equally to this workCorrespondence: Ying WangDepartment of Cardiovascular Medicine, Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine and Science, 200 1st ST SW, Rochester, MN, 55905, USAEmail [email protected]: Endothelial cell (EC) activation facilitates leukocyte adhesion to vascular walls, which is implicated in a variety of cardiovascular diseases and is a target for prevention and treatment. Despite the development of anti-inflammatory medications, cost-effective therapies with significant anti-inflammatory effects and lower organ toxicity remain elusive. The goal of this study is to identify novel synthetic compounds that inhibit EC inflammatory response with minimal organ toxicity.Methods and Results: In this study, we discovered LCC-09, a salicylanilide derivative consisting of the functional fragment of magnolol, 2,4-difluorophenyl, and paeonol moiety of salicylate, as a novel anti-inflammatory compound in cultured ECs and zebrafish model. LCC-09 was shown to inhibit pro-inflammatory cytokine tumor necrosis factor-α (TNFα)-induced expression of adhesion molecules and inflammatory cytokines, leading to reduced leukocyte adhesion to ECs. Mechanistically, LCC-09 inhibits the phosphorylation of signal transducer and activator of transcription 1 (STAT1), TNFα-induced degradation of NF-κ-B Inhibitor-α (IκBα) and phosphorylation of NFκB p65, resulting in reduced NFκB transactivation activity and binding to E-selectin promoter. Additionally, LCC-09 attenuated TNFα-induced generation of reactive oxygen species in ECs. Molecular docking models suggest the binding of LCC-09 to NFκB essential modulator (NEMO) and Janus tyrosine kinase (JAK) may lead to dual inhibition of NFκB and STAT1. Furthermore, the anti-inflammatory effect of LCC-09 was validated in the lipopolysaccharides (LPS)-induced inflammation model in zebrafish. Our results demonstrated that LCC-09 significantly reduced the LPS-induced leukocyte recruitment and mortality of zebrafish embryos. Finally, LCC-09 was administered to cultured ECs and zebrafish embryos and showed minimal toxicities.Conclusion: Our results support that LCC-09 inhibits EC inflammatory response but does not elicit significant toxicity.Keywords: endothelial cells, inflammation, salicylanilide derivative, tumor necrosis factor-α, lipopolysaccharides, toxicity

Keywords

• our results support that lcc-09 inhibits ec inflammatory response but does not elicit significant toxicity.