Triple helix-forming oligonucleotides conjugated to new inhibitors of topoisomerase II: synthesis and binding properties.
Triplex-forming oligonucleotides (TFOs) are among the most specific DNA ligands and represent an important tool for specific regulation of gene expression. TFOs have also been used to target DNA-modifying molecules to obtain irreversible modifications on a specific site of the genome. A number of molecules have been recognized to target topoisomerase II and stabilize double-stranded cleavage mediated by this enzyme thus determining permanent DNA damage. Among these poisons, etoposide (VP16), a 4'-demethylepipodophyllotoxin derivative, is widely used in cancer chemotherapy. In the aim to design DNA site-specific molecules, three analogues of VP16 (1, 2, and 3), recently described (Duca et al. J. Med. Chem. 2005, 48, 596-603), were attached to TFOs, together with a fourth one, of which the synthesis is reported here. Two different oligonucleotides, differing by the length (a 16-mer and a 20-mer), and two different linker arms between the oligonucleotide and the drug were used. The coupling reaction between the drug and the TFO was further improved. For the first time, we also report the synthesis of TFO conjugates bearing two molecules of inhibitor linked to the same oligonucleotide end. In total, 16 new conjugates were synthesized and evaluated for their ability to form triple helices. The loss in triplex stability due to the conjugation of the TFO to compounds that do not interact with DNA is compensated by the presence of the ethylene glycol linker arm. This stabilization effect is more pronounced at the 3' end than at the 5' end. All conjugates form a stable triplex selectively on the DNA target at 37 degrees C and pH 7.2.
Duca
Maria
M
Muséum National d'Histoire Naturelle, USM0503 MNHN, UMR5153 CNRS, U565 INSERM, 43 rue Cuvier CP26 75231 Paris Cedex 05 France.
Oussedik
Kahina
K
Ceccaldi
Alexandre
A
Halby
Ludovic
L
Guianvarc'h
Dominique
D
Dauzonne
Daniel
D
Monneret
Claude
C
Sun
Jian-Sheng
JS
Arimondo
Paola B
PB
D004247:DNA / Q000378:metabolism; D018983:DNA Footprinting; D004791:Enzyme Inhibitors / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008958:Models, Molecular; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D059005:Topoisomerase II Inhibitors
D004791:Enzyme Inhibitors; D009841:Oligonucleotides; D059005:Topoisomerase II Inhibitors; D004247:DNA