Novel Synthesized N-Ethyl-Piperazinyl-Amides of C2-Substituted Oleanonic and Ursonic Acids Exhibit Cytotoxic Effects through Apoptotic Cell Death Regulation.

Oxana Kazakova,George Drăghici,Irina Smirnova,Marius Mioc,Adrian Voicu,Cristina Dehelean,Alexandra Mioc,Ştefana Avram,Lavinia Vlaia,Irina Baikova,Codruța Şoica,Ioana Macașoi

doi:10.3390/ijms222010967

Abstract

A series of novel hybrid chalcone N-ethyl-piperazinyl amide derivatives of oleanonic and ursonic acids were synthesized, and their cytotoxic potential was evaluated in vitro against the NCI-60 cancer cell line panel. Compounds 4 and 6 exhibited the highest overall anticancer activity, with GI50 values in some cases reaching nanomolar values. Thus, the two compounds were further assessed in detail in order to identify a possible apoptosis- and antiangiogenic-based mechanism of action induced by the assessed compounds. DAPI staining revealed that both compounds induced nuclei condensation and overall cell morphological changes consistent with apoptotic cell death. rtPCR analysis showed that up-regulation of pro-apoptotic Bak gene combined with the down-regulation of the pro-survival Bcl-XL and Bcl-2 genes caused altered ratios between the pro-apoptotic and anti-apoptotic proteins’ levels, leading to overall induced apoptosis. Molecular docking analysis revealed that both compounds exhibited high scores for Bcl-XL inhibition, suggesting that compounds may induce apoptotic cell death through targeted anti-apoptotic protein inhibition, as well. Ex vivo determinations showed that both compounds did not significantly alter the angiogenesis process on the tested cell lines.

Highlights

The present study reports the anticancer biological assessment of a series of novel synthesized ursonic and oleanonic acids’ derivatives
These compounds were screened against the NCI-60 cancer cell line panel
By means of DAPI staining, it was determined that the tested compounds affected overall cell and nuclei morphology

Summary

Introduction

Malignant diseases have always been present in the history of mankind; the name “cancer” is related to the Greek physician Hippocrates (460–370 BC), who used the term “carcinoma” to describe malignant tumors, a term translated by the Roman physician. Celsus (28–50 BC) into the Latin word “cancer”. Both terms mean crab, probably due to the ramified aspect of tumors [1]. The incidence of cancer cases has increased in the modern age, most likely due to the significant increase of life expectancy [2]. Nowadays, it counts as the second leading cause of death globally [3].

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Conclusion