New 3-Aryl-2-(2-thienyl)acrylonitriles with High Activity Against Hepatoma Cells

Abstract

New 2-(thien-2-yl)-acrylonitriles with putative kinase inhibitory activity were prepared and tested for their antineoplastic efficacy in hepatoma models. Four out of the 14 derivatives were shown to inhibit hepatoma cell proliferation at (sub-)micromolar concentrations with IC50 values below that of the clinically relevant multikinase inhibitor sorafenib, which served as a reference. Colony formation assays as well as primary in vivo examinations of hepatoma tumors grown on the chorioallantoic membrane of fertilized chicken eggs (CAM assay) confirmed the excellent antineoplastic efficacy of the new derivatives. Their mode of action included an induction of apoptotic capsase-3 activity, while no contribution of unspecific cytotoxic effects was observed in LDH-release measurements. Kinase profiling of cancer relevant protein kinases identified the two 3-aryl-2-(thien-2-yl)acrylonitrile derivatives 1b and 1c as (multi-)kinase inhibitors with a preferential activity against the VEGFR-2 tyrosine kinase. Additional bioinformatic analysis of the VEGFR-2 binding modes by docking and molecular dynamics calculations supported the experimental findings and indicated that the hydroxy group of 1c might be crucial for its distinct inhibitory potency against VEGFR-2. Forthcoming studies will further unveil the underlying mode of action of the promising new derivatives as well as their suitability as an urgently needed novel approach in HCC treatment.

Anticancer Activity and Mechanisms of Action of New Chimeric EGFR/HDAC-Inhibitors

“…To exclude unspecific cytotoxicity as the driving mode of action for antiproliferative effects, the release of lactate dehydrogenase (LDH) from DU145 and Hep-G2 cells was determined after 3 h and 24 h of treatment using the Cytotoxicity Detection Kit PLUS LDH (Roche Diagnostics GmbH, Mannheim, Germany). The assay was performed as described earlier [37]. Adherent non-treated cells, lysed with 2% Triton X-100 media for 10 min, served as reference values for maximum LDH release.…”

“…Changes in caspase-3 activity were measured by the cleavage of the fluorogenic substrate AC-DEVD-AMC (EMD Millipore, Billerica, MA, USA), as described previously [37]. After incubation with 3BrQuin-SAHA, 3ClQuin-SAHA, SAHA, or gefitinib (1-10 µM) for 6-48 h, the cells were harvested and lysed with lysis buffer.…”

Insights into the chemistry and therapeutic potential of acrylonitrile derivatives

“…The authors argued that the formation of a stable complex of 15 and VEGFR‐2 as a result of the molecular docking and dynamic studies supported their biological results. [ 121 ]…”

Improved Anticancer Activities of a New Pentafluorothio-Substituted Vorinostat-Type Histone Deacetylase Inhibitor

“…To exclude unspecific cytotoxicity as the driving mode of action for antiproliferative effects, the release of lactate dehydrogenase (LDH) from DU145 cells was determined after 3 h and 24 h of treatment using the Cytotoxicity Detection Kit PLUS LDH (Roche Diagnostics GmbH, Mannheim, Germany). The assay was performed as described earlier [43]. Cytotoxicity was determined by subtracting the percentage of LDH release into the supernatant under control conditions of those from treated samples.…”

“…Changes in caspase-3 activity were measured by the cleavage of the fluorogenic substrate AC-DEVD-AMC (EMD Millipore, Billerica, MA, USA), as described previously [43]. After incubation with SF5-SAHA or SAHA (1-10 µM) for 24 h the cells were harvested and lysed with lysis buffer.…”