1600P - Phospho-kinase expression and anti-tumoral activity of the novel multikinase inhibitor EC-70124 in colon cancer

Date 28 September 2014
Event ESMO 2014
Session Poster Display session
Topics Drug Development
Colon Cancer
Translational Research
Presenter Alberto Ocana
Citation Annals of Oncology (2014) 25 (suppl_4): iv546-iv563. 10.1093/annonc/mdu358
Authors A. Ocana1, M. Cuenca1, G. Serrano2, J.C. Montero3, V. Corrales4, F. Morís5, L.E. Nuñez5, A. Pandiella3
  • 1Translational Oncology, Research Unit Clinical Hospital Albacete, 02006 - Albacete/ES
  • 2Molecular Biology, Research Unit Clinical Hospital Albacete, Albacete/ES
  • 3Cancer Research Center,, University of Salamanca, salamanca/ES
  • 4Translational Oncology, Research Unit Clinical Hospital Albacete, Albacete/ES
  • 5Entrechem Sl, Universidad de Oviedo, Oviedo/ES

Abstract

Aim

Protein kinases are widely activated in different solid tumors including colorectal cancer. Therapeutic inhibition of some of them has shown clinical benefit. In the present study we describe the anti-tumoral activity of the novel multikinase inhibitor EC-70124 based on the kinase profile observed in colon cancer samples.

Methods

phospho-kinase array kits were used for the evaluation of activated protein kinases in human samples from colon cancer patients. A panel of cell lines was used to confirm the kinase profile observed in patients including: SW48, SW620 and HT29. Western blot analyses were performed to evaluate the activated proteins. Cell proliferation and growth was measured by MTT uptake and matrigel cultures. Evaluation of apoptosis and cell cycle was performed by flow cytometry using Annexin V and propidium iodide, respectively. To evaluate if EC-70124 combined with other chemotherapy drugs was synergistic, additive, or antagonist, we used the CalcuSyn v2.0 software program.

Results

18 patients were evaluated for the expression of phospho kinases. Most frequent activated kinases included ErbB receptors, the VEGFR family and FGFR. Mediators of downstream routes included AKT, S6, STAT1 and ALK. EC-70124 inhibited components of the PI3K/AKT/mTOR including AKT and pS6 in addition to SRC. Treatment with EC-70124 reduced proliferation and cell growth in SW620 and HT29, in addition to cell migration. Flow cytometry analyses showed an arrest at the G2/M phase and an induction of apoptosis. EC-70124 at 500nM led to an induction of DNA damage measured by ɣH2AX and a processing of PARP. EC-70124 in combination with chemotherapy increased its activity through an induction of apoptosis.

Conclusions

EC-70124 shows antitumor activity in colon cancer cell lines acting on relevant activated pathways observed in human samples.

Disclosure

All authors have declared no conflicts of interest.