Synthesis, functional proteomics and biological evaluation of new 5-pyrazolyl ureas as potential anti-angiogenic compounds

• Published in: European journal of medicinal chemistry Vol. 226; p. 113872
• Main Authors: Morretta, Elva, Sidibè, Adama, Spallarossa, Andrea, Petrella, Antonello, Meta, Elda, Bruno, Olga, Monti, Maria Chiara, Brullo, Chiara
• Format: Journal Article
• Language: English
• Published: France Elsevier Masson SAS 15-12-2021
• Subjects: 5-Pyrazolyl ureas; Angiogenesis; Angiogenesis Inhibitors - chemical synthesis; Angiogenesis Inhibitors - chemistry; Angiogenesis Inhibitors - pharmacology; Dose-Response Relationship, Drug; Drug affinity responsive target stability; Functional proteomics; Humans; Limited proteolysis; Molecular Structure; Neovascularization, Physiologic - drug effects; Protein Phosphatase 1 - antagonists & inhibitors; Protein Phosphatase 1 - metabolism; Protein-ligand interactions; Proteolysis - drug effects; Pyrazoles - chemical synthesis; Pyrazoles - chemistry; Pyrazoles - pharmacology; Serine/threonine-protein phosphatase PP1γ; Structure-Activity Relationship; Urea - analogs & derivatives; Urea - chemistry; Urea - pharmacology; Angiogenesis; Protein-ligand interactions; 5-Pyrazolyl ureas; Functional proteomics; Serine/threonine-protein phosphatase PP1γ; Drug affinity responsive target stability; Limited proteolysis
• ISSN: 0223-5234; 1768-3254
• DOI: 10.1016/j.ejmech.2021.113872

Based on biological results of previous synthesized pyrazolyl ureas able to interfere with angiogenesis process, we planned and synthesized the new benzyl-urea derivatives 2–4; some of them showed an interesting anti-proliferative profile and particularly 4e potently inhibited HUVEC proliferation. To shed light on the mechanism of action of 4e, its interactome has been deeply inspected to identify the most prominent protein partners, mainly taking into account kinome and phosphatome, through drug affinity responsive target stability experiments, followed by targeted limited proteolysis analysis. From these studies, PP1γ emerged as the most reliable 4e potential target in HUVEC. Molecular docking simulations on PP1γ were carried out to predict 4e binding mode. To assess its potential anti-angiogenic effect, 4e was tested in vitro to verify interference on kinase and phosphate activities. Overall, our results evidenced for 4e an interesting anti-angiogenic action, probably due to its action at intracellular level on PP1γ signalling pathways.
[Display omitted]
•New derivatives 2–4 were designed and synthesized, showing good proliferation inhibition of cancer and HUVEC cells.•The interactome of 4e was investigated by DARTS and t-LiP experiments to identify its target(s).•PP1γ emerged as possible target protein involved in intracellular mechanism of action of 4e in HUVEC cells.•Molecular docking simulation on PP1γ was performed to verify a possible interaction.•Compound 4e showed potential anti-angiogenic action through PP1γ signalling pathways.