TY - JOUR
T1 - Discovery of anticancer agents with c-Met inhibitory potential by virtual and experimental screening of a chemical library
AU - Mortazavi, Motahareh
AU - Raufi, Elaheh
AU - Damghani, Tahereh
AU - Khoshneviszadeh, Mehdi
AU - Edraki, Najmeh
AU - Eskandari, Masoomeh
AU - Giovannetti, Elisa
AU - Peters, Godefridus J.
AU - Pirhadi, Somayeh
AU - Firuzi, Omidreza
N1 - Funding Information: This study was supported by the National Institute for Medical Research Development (NIMAD , Grant number: 957652 ). We also wish to thank the Vice-Chancellor for Research, Shiraz University of Medical Sciences. Part of this study was the Pharm. D. thesis project of Elaheh Raufi at Shiraz University of Medical Sciences (Thesis number: 1152). Funding Information: This study was supported by the National Institute for Medical Research Development (NIMAD, Grant number: 957652). We also wish to thank the Vice-Chancellor for Research, Shiraz University of Medical Sciences. Part of this study was the Pharm. D. thesis project of Elaheh Raufi at Shiraz University of Medical Sciences (Thesis number: 1152). Publisher Copyright: © 2022
PY - 2023/1/5
Y1 - 2023/1/5
N2 - c-Met receptor tyrosine kinase has recently emerged as an important target with therapeutic implications in pancreatic cancer. In this study, we carried out a docking virtual screening on an in-house library of 441 synthesized compounds and selected the compounds with the best interactions with the c-Met protein to be subjected to experimental tests. Ten compounds belonging to 3 different classes of chemical structures were selected for this purpose and their antiproliferative effects were studied against 4 pancreatic ductal adenocarcinoma (PDAC) cell lines including AsPC-1, Suit-2, Panc-1 and Mia-Paca-2 cells, primary PDAC cells and also c-Met amplified EBC-1 cell line by sulforhodamine-B assay. Apoptosis induction was examined by Hoechst 33258 staining and annexin V-FITC/propidium iodide flow cytometric assay. The best compound was also assayed in three-dimensional cultures of AsPC-1 cells and its c-Met inhibitory potential was studied by immunoblotting and a homogenous time resolved fluorescence (HTRF) assay. The compound with a phenanthrotriazine hydrazinyl scaffold bearing nitrophenyl pendant (PhTH) was the most active derivative, with IC50 values in the range of 5–8 μM. This compound exerted antiproliferative effect against AsPC-1 cells also in the presence of hepatocyte growth factor (HGF). PhTH induced apoptosis, dose-dependently inhibited spheroid growth, inhibited c-Met activity in cell-free HTRF assay and also inhibited the phosphorylation of c-Met and its downstream effector ERK1/2 in AsPC-1 cells. Molecular docking and dynamics simulation and MM-PBSA analysis confirmed close interactions of PhTH with c-Met kinase domain. Some of the tested compounds in this study seem to be potential c-Met inhibitors with promising activities against PDAC cells.
AB - c-Met receptor tyrosine kinase has recently emerged as an important target with therapeutic implications in pancreatic cancer. In this study, we carried out a docking virtual screening on an in-house library of 441 synthesized compounds and selected the compounds with the best interactions with the c-Met protein to be subjected to experimental tests. Ten compounds belonging to 3 different classes of chemical structures were selected for this purpose and their antiproliferative effects were studied against 4 pancreatic ductal adenocarcinoma (PDAC) cell lines including AsPC-1, Suit-2, Panc-1 and Mia-Paca-2 cells, primary PDAC cells and also c-Met amplified EBC-1 cell line by sulforhodamine-B assay. Apoptosis induction was examined by Hoechst 33258 staining and annexin V-FITC/propidium iodide flow cytometric assay. The best compound was also assayed in three-dimensional cultures of AsPC-1 cells and its c-Met inhibitory potential was studied by immunoblotting and a homogenous time resolved fluorescence (HTRF) assay. The compound with a phenanthrotriazine hydrazinyl scaffold bearing nitrophenyl pendant (PhTH) was the most active derivative, with IC50 values in the range of 5–8 μM. This compound exerted antiproliferative effect against AsPC-1 cells also in the presence of hepatocyte growth factor (HGF). PhTH induced apoptosis, dose-dependently inhibited spheroid growth, inhibited c-Met activity in cell-free HTRF assay and also inhibited the phosphorylation of c-Met and its downstream effector ERK1/2 in AsPC-1 cells. Molecular docking and dynamics simulation and MM-PBSA analysis confirmed close interactions of PhTH with c-Met kinase domain. Some of the tested compounds in this study seem to be potential c-Met inhibitors with promising activities against PDAC cells.
KW - Anticancer agents
KW - Computational screening
KW - Gastrointestinal tumors
KW - Kinase inhibitors
KW - Targeted therapies
UR - http://www.scopus.com/inward/record.url?scp=85143162338&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.ejphar.2022.175395
DO - https://doi.org/10.1016/j.ejphar.2022.175395
M3 - Article
C2 - 36410418
SN - 0014-2999
VL - 938
JO - European journal of pharmacology
JF - European journal of pharmacology
M1 - 175395
ER -