Coralie Caron (2019)

Macrocyclic ligands for DNA abasic sites as inhibitors of DNA repair : Synthesis, biochemical and biological studies

Abstract

In the context of chemotherapy, DNA repair reduces the DNA damage induced by DNA-alkylating drugs such as temozolomide, leading to chemoresistance. One of the most important pathways of DNA repair is Base Excision Repair (BER), where a key enzyme, APE1 (AP endonuclease 1), cleaves abasic sites generated following treatment with DNA-alkylating drugs and initiates the repair of the single-strand break. The DNA repair activity of APE1 was identified as the major source of chemoresistance in certain cancers. Several studies validated the BER pathway and, particularly, the APE1 enzyme as important drug targets for improvement the efficacy of anti-cancer drugs; for this reason, several APE1 inhibitors have been developed. However, instead of direct inhibition of the enzyme, an alternative strategy can rely on targeting its substrate: the AP sites in DNA. Macrocyclic compounds, namely naphthalenophanes, show a strong and selective binding to abasic sites in the DNA. This process interferes with the recognition of the latter by APE1 and leads in vitro to two effects: inhibition of the APE1-induced DNA cleavage and macrocycle-induced DNA cleavage by a mechanism different from that of APE1, namely β-elimination. Herein, a novel serie of functionalized naphthalenophanes, composed of nine novel derivatives, has been synthesized and studied. Most ligands demonstrate a strong and selective binding to AP-sites in DNA and an inhibition of APE1 activity in vitro, with inhibitory constants from 39 nM to 25 µM. Moreover, the inhibitory activity of ligands, as characterized by Kı values, could be directly related to their affinity and selectivity to AP-sites. The molecular design of macrocycles has a crucial influence on their intrinsic AP-site cleavage activity leading either to total abolition, or to an exceptionally high AP-site cleavage activity. Interestingly, an unprecedented formation of a covalent DNA-ligand adduct with one of the ligands have been characterized. Finally, the biological activity of naphthalenophanes was assessed in the TMZ-resistant glioblastoma cell line T98G. Most compounds are highly active, with GI₅₀ values in sub-micromolar or low-micromolar range. In addition, a remarkable synergic effect upon co-treatment of TMZ or MMS with one ligand (2,7-BisNP-O4Me) was demonstrated. This ligand was found to increase the number of AP-sites and the number of double-strands break in DNA upon co-treatment with TMZ and MMS suggesting APE1 inhibition as excepted. These observations highlight the hight therapeutic interest of this compound. (Defended on October 18, 2019)