RESUMEN
The synthesis and evaluation for telomerase-inhibitory and quadruplex DNA binding properties of three related series of rationally designed trisubstituted acridine derivatives are described. These are substituted on the acridine ring at the 2,6,9; 2,7,9; and 3,6,9 positions. The ability of several of the most potent compounds to interact with and stabilize an intramolecular G-quadruplex DNA was evaluated by surface plasmon resonance methods, and affinities were found to correlate with potency in a telomerase assay. The interactions of a number of compounds with a parallel quadruplex DNA structure were simulated by molecular modeling methods. The calculated interaction energies were compared with telomerase activity and showed generally consistent correlations between quadruplex affinity and telomerase inhibition. These data support a model for the action of these compounds that involves the stabilization of intermediate quadruplex structures that inhibit the elongation of telomeric DNA by telomerase in tumor cells.
Asunto(s)
Acridinas/síntesis química , Acridinas/farmacología , Inhibidores de la Transcriptasa Inversa/síntesis química , Inhibidores de la Transcriptasa Inversa/farmacología , Telomerasa/antagonistas & inhibidores , Cristalografía por Rayos X , ADN/efectos de los fármacos , ADN/metabolismo , ADN de Neoplasias/biosíntesis , ADN de Neoplasias/metabolismo , Humanos , Cinética , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Conformación Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rodaminas/química , Relación Estructura-Actividad , Resonancia por Plasmón de Superficie , Células Tumorales CultivadasRESUMEN
[reaction: see text] A novel high-yielding method for the solid-phase synthesis of 3,6-bispeptide-acridone conjugates is reported. It involves initial coupling of bifunctionalized acridone to a resin-bound peptide followed by an on-bead site-site reaction to couple the second peptide. This method leads to clean symmetrical bispeptide derivatives and appears to be general. This strategy will enable the generation of a library of 3,6-bispeptide-acridones to be screened for selective binding to telomeric G-quadruplex DNA.
Asunto(s)
Acridinas/síntesis química , Técnicas Químicas Combinatorias/métodos , Dipéptidos/síntesis química , Acridinas/química , Acridonas , Antineoplásicos/síntesis química , Antineoplásicos/química , ADN/antagonistas & inhibidores , Dipéptidos/química , Diseño de Fármacos , G-Cuádruplex , Ligandos , Imitación Molecular , Biblioteca de Péptidos , Telómero/químicaRESUMEN
The synthesis and evaluation of a group of 2,6-, 2,7- and 3,6-bis-aminoalkylamido acridones are reported, which show a similar level of activity against telomerase in vitro compared to their acridine counterparts. Computer modelling and calculations of relative binding energies suggest an equivalent binding mode to human intramolecular G-quadruplex DNA, but with significantly reduced affinity, as a result of the limited delocalisation of the acridone chromophore compared to the acridine system. Thermal melting studies on acridone and acridine quadruplex complexes using a FRET approach support these predictions. Long-term cell proliferation studies at sub-cytotoxic doses with two representative acridones using the SKOV3 cell line, show that neither compound produces growth arrest, in contrast with the effects produced by the tri-substituted acridine compound BRACO-19. It is concluded that telomerase inhibitory activity is a necessary though by itself insufficient property in order for cellular growth arrest to occur at sub-toxic concentrations, and that tight quadruplex binding is also required.