A Guanidine-Based Synthetic Compound Suppresses Angiogenesis via Inhibition of Acid Ceramidase

Abstract

Angiogenesis generates new blood vessels from pre-existing vessels. Tumors induce the formation of new blood vessels to ensure sufficient oxygen and nutrients for their growth. Normally, angiogenesis is induced by various pro-angiogenesis factors, including vascular endothelial growth factor (VEGF). Inhibition of VEGF is a promising approach to cancer treatment. A guanidine-based synthetic compound, E2, was identified as a potent hit from 68 guanidine-based derivatives by screening for angiogenesis inhibitors showing antiproliferative activity in human umbilical vein endothelial cells (HUVECs). To explore the mode of action of E2, target proteins were investigated using phage display biopanning, and acid ceramidase 1 (ASAH1) was identified as an E2-binding protein. Drug affinity responsive target stability (DARTS) and ASAH1 activity assays revealed the direct binding of E2 to ASAH1. Moreover, siRNA knockdown of ASAH1 demonstrated its role as an angiogenesis factor. Consequently, E2 inhibited chemoinvasion and tube formation of HUVECs in a dose-dependent manner. E2 also potently suppressed neo-vascularization of chorioallantoic membranes in vivo. Collectively, these data suggest that E2 is a novel angiogenesis inhibitor and ASAH1 is proposed to be a new antiangiogenesis target.

Profiling the Protein Targets of Unmodified Bio‐Active Molecules with Drug Affinity Responsive Target Stability and Liquid Chromatography/Tandem Mass Spectrometry

“…From phage display biopanning, the E2 binding protein acid ceramidase 1 (ASAH1) was identified. The DARTS assay revealed that E2 increased the stability of ASAH1 against proteolysis with pronase . The binding of E2 to ASAH1 leads to suppression of vascular endothelial growth factor through the Akt signaling pathway, thus leading to inhibition of angiogenesis.…”

“…DARTS assays of unmodified drugs can be combined with immunoblotting to validate the DARTS and LC‐MS/MS results and investigate the biophysical interactions of bioactive small molecules with target proteins . Alteration in the stability of the target protein by a bioactive small molecule after treatment with pronase indicates that there was a direct biophysical interaction between the two molecules.…”

Reactions of thiourea oxides with amines, aminoacids and proteins

“…It should be noted in conclusion, that chemists continue to use thiourea oxides in the synthesis of guanidine-based pharmaceuticals. Thus, in 2019 Cho and coworkers have received guanidine-based compound which suppresses angiogenesis in human umbilical vein ebdothelial cells [26]. In the intermediate steps of the synthesis of this compound they used thiourea trioxide.…”

Elusive Roles of the Different Ceramidases in Human Health, Pathophysiology, and Tissue Regeneration

“…ACDase also regulates cancer cell motility through the activation of the ITGαVβ5/FAK signaling cascade [ 63 ]. Additionally, the significant roles of ACDase in angiogenesis, chronic inflammation, and tumorigenesis may contribute to cancer development and progression [ 66 , 67 ]. ACDase affects multiple factors in cancer pathogenicity, which adds to the complexity of the enzyme in the diagnosis and treatment of the disease.…”