A novel inhibitor, 16-hydroxy-cleroda-3,13-dien-16,15-olide, blocks the autophosphorylation site of focal adhesion kinase (Y397) by molecular docking

• Published in: Biochimica et biophysica acta Vol. 1830; no. 8; pp. 4091 - 4101
• Main Authors: Thiyagarajan, Varadharajan, Lin, Shih-Hung, Chia, Yi-Chen, Weng, Ching-Feng
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.08.2013
• Subjects: 16-hydroxy-cleroda-3,13-dien-16,15-olide; Animals; Anti-tumorigenesis; Antineoplastic Agents - pharmacology; apoptosis; Apoptosis - drug effects; atomic force microscopy; catechin; Cell Cycle - drug effects; Cell Line, Tumor; cell movement; cell viability; curcumin; Diterpenes - pharmacology; energy; Epithelial mesenchymal transition; Filopodia; Focal adhesion kinase; Focal Adhesion Protein-Tyrosine Kinases - antagonists & inhibitors; Mice; Models, Molecular; Molecular Docking Simulation; molecular models; non-specific protein-tyrosine kinase; Phosphorylation; pro-apoptotic proteins; Protein Kinase Inhibitors - pharmacology; proteins; pseudopodia; quercetin; Rats; signal transduction; Signal Transduction - drug effects; tyrosine; Anti-tumorigenesis; Focal adhesion kinase; Epithelial mesenchymal transition; 16-hydroxy-cleroda-3,13-dien-16,15-olide; Filopodia
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2013.04.027

Focal adhesion kinase (FAK) is a nonreceptor protein tyrosine plays an important role in a number of cell signaling pathways, including cell migration, proliferation, and cell survival. This study was aimed to identify novel and specific inhibitors from natural compounds via molecular docking of FAK (Y397). The 3D structure of FAK (PDB ID: 2AL6) was used for docking 109 natural compounds. Based on high affinity and energy interaction, four of ten candidate compounds, 16-hydroxy-cleroda-3,13-dien-16,15-olide (HCD), curcumin, quercetin, and catechin hydrate, were hit, and the inhibitory activity against FAK was validated in these compounds in C6 glioma and N18 neuroblastoma cell lines. HCD showed a potential effect on cell viability by MTT assay and cell arrest in the G0–G1 phase, and a TUNEL assay confirmed further apoptosis. Treatment with HCD decreased anti-apoptotic proteins and increased pro-apoptotic proteins. Atomic force microscopy data depicted that the formation of filopodia on the intracellular surface decreased in treated cells compared with the control. Zymography showed that HCD inhibited the activity of MMP-2 and MMP-9. The protein levels of FAK, pFAK, Rac1 and Cdc42, which are the key regulators for the formation of filopodia, were decreased. Additionally, HCD regulated the expression of epithelial mesenchymal transition proteins. HCD effectively interacted at the autophosphorylation site of FAK and interaction analysis indicated an H-bond with the Arg 86 and Arg 125 residues. This study suggests that HCD could be a potential inhibitor of FAK and could be used for anti-tumorigenesis and anti-metastasis treatments. •HCD blocked the autophosphorylation site of FAK.•HCD regulated the expression of epithelial mesenchymal transition proteins.•HCD decreased the formation of filopodia and induce the apoptosis.