RESUMEN
We have prepared oligonucleotides with a naphthylquinoline triplex-binding ligand covalently tethered to the 5'-end and have used UV-melting and DNase I footprinting to examine the stability of intra- and inter-molecular triplexes containing this modification. We find that covalent attachment of the ligand increases the melting temperature of intramolecular 6-mer triplexes by about 14 K, and increases the binding of 9-mer oligonucleotides to their duplex target sites by about 60-fold.
Asunto(s)
ADN/química , Conformación de Ácido Nucleico , Quinolinas/química , Animales , Bovinos , Huella de ADN , Desoxirribonucleasa I , LigandosRESUMEN
On the basis of a systematic SAR analysis of substituted quinolines, a derivative 32 was synthesized that shows half-maximal inhibition of the immunostimulatory effect of CpG-oligodeoxynucleotides in vitro at the concentration of 0.24 nM.
Asunto(s)
Aminoquinolinas/síntesis química , Aminoquinolinas/farmacología , Oligonucleótidos/antagonistas & inhibidores , Cloroquina/análogos & derivados , Islas de CpG/inmunología , Cinética , Relación Estructura-ActividadRESUMEN
We have used DNase I footprinting to examine the effect of a novel naphthylquinoline dimer, designed as a triplex-specific bis-intercalator, on the stability of intermolecular DNA triplexes. We find that this compound efficiently promotes triplex formation between the 9-mer oligonucleotide 5'-TTTTTTCTT and its oligopurine duplex target at concentrations as low as 0.1 microM, enhancing the triplex stability by at least 1000-fold. This compound, which is the first reported example of a triplex bis-intercalator, is about 30 times more potent than the simple monofunctional ligand.