Antitumorigenic Effects of Inhibiting Ephrin Receptor Kinase Signaling by GLPG1790 against Colorectal Cancer Cell Lines In Vitro and In Vivo

Abstract

Erythropoietin-producing hepatocellular receptors (Eph) promote the onset and sustain the progression of cancers such as colorectal cancer (CRC), in which the A2 subtype of Eph receptor expression has been shown to correlate with a poor prognosis and has been identified as a promising therapeutic target. Herein, we investigated, in vitro and in vivo, the effects of treatment with GLPG1790, a potent pan-Eph inhibitor. The small molecule has selective activity against the EphA2 isoform in human HCT116 and HCT15 CRC cell lines expressing a constitutively active form of RAS concurrently with a wild-type or mutant form of p53, respectively. GLPG1790 reduced EPHA2 phosphorylation/activation and induced G1/S cell-cycle growth arrest by downregulating the expression of cyclin E and PCNA, while upregulating p21Waf1/Cip1 and p27Cip/Kip. The inhibition of ephrin signaling induced quiescence in HCT15 and senescence in HCT116 cells. While investigating the role of CRC-related, pro-oncogenic p53 and RAS pathways, we found that GLPG1790 upregulated p53 expression and that silencing p53 or inhibiting RAS (human rat sarcoma)/ERKs (extracellular signal-regulated kinase) signaling restrained the ability of GLPG1790 to induce senescence in HCT116 cells. On the other hand, HCT15 silencing of p53 predisposed cells to GLPG1790-induced senescence, whilst no effects of ERK inhibition were observed. Finally, GLPG1790 hindered the epithelial-mesenchymal transition, reduced the migratory capacities of CRC, and affected tumor formation in xenograft models in vivo more efficiently using HCT116 than HCT15 for xenografts. Taken together, our data suggest the therapeutic potential of GLPG1790 as a signal transduction-based therapeutic strategy in to treat CRC.

Optimization of the Lead Compound NVP‐BHG712 as a Colorectal Cancer Inhibitor

“…However, shifting the residues to the 4-position resulted in significantly less EPHA2 affine derivatives (15)(16)(17)(18), and by introduction of larger residues like morpholine (23) and COOEt (24) the affinities were even further reduced. In contrast to this finding, derivatives featuring the optimized disubstituted Chemistry-A European Journal phenylbenzamide substructures from Nilotinib and Ponatinib were well tolerated, showing high binding affinities towards EPHA2 (8, K D = 33 nM and 9, K D = 5 nM).…”

“…Initial studies in CRC models showed promising effects of EPHA2 inhibition by small molecules. Specifically, the potent pan‐EPH inhibitor GLPG1790 was shown to selectively target EPHA2 by reducing its phosphorylation/activation in human CRC cells and inducing cell‐cycle arrest [24] . Furthermore, ALW‐II‐41‐27, a specific EPHA2 inhibitor, was shown to revert acquired resistance to the anti‐EGFR antibody Cetuximab in human CRC models [25] …”

“…Specifically, the potent pan-EPH inhibitor GLPG1790 was shown to selectively target EPHA2 by reducing its phosphorylation/activation in human CRC cells and inducing cell-cycle arrest. [24] Furthermore, ALW-II-41-27, a specific EPHA2 inhibitor, was shown to revert acquired resistance to the anti-EGFR antibody Cetuximab in human CRC models. [25] Previously, we performed a broad off-target analysis of preclinically and clinically investigated kinase inhibitors using chemical proteomics (Kinobeads technology), NMR-based conformational dynamics and crystal structures of the EPHA2inhibitor complexes.…”

Metabolic Reprogramming in Metastatic Melanoma with Acquired Resistance to Targeted Therapies: Integrative Metabolomic and Proteomic Analysis

“…As EPHA2 is a modulator of the MAPK pathway, its early inhibition may delay the development of drug resistance. Such inhibitors have already shown an antitumoral effect in vitro and in vivo in colorectal and lung cancers [64,65].…”

The Importance of Tumor Stem Cells in Glioblastoma Resistance to Therapy

“…As for other cancers [ 43 ], Eph (erythropoietin-producing hepatocellular carcinoma) receptor family members are reported as key molecules in GBM development. These receptors are primarily expressed in early development and are crucial for embryonic development and regulating processes such as cell adhesion and migration [ 44 ].…”