NMR in drug discovery. From screening to structure-based design of antitumoral agents

摘要: Nuclear Magnetic Resonance has experienced an increasing interest in the drug discovery field that led to its wide use on nearly every stage of development. For this reason, during present thesis we propose some tools offered by NMR target various systems related with cancer. Initially intended get acquainted most outstanding methodologies for detection and characterization binding events; goal proteins involved cellular apoptosis (XIAP Bcl-XL) were used set up both ligand receptor based experiments. First developed two novel labeling selective observation Trp, which coupled receptor-based experiments provided insight XIAP-ligand complexes, as well significant advantages HTS. Later, such STD, WaterLOGSY iLOE applied screening weak binders Bcl-XL, resulting information reconstruction dual a fragment-based approach. In second same development protein-protein interaction inhibitors, particular pro-angiogenic protein VEGF; end different strategies proposed. First, small diverse library D-oligopeptides was screened ligands using former strategy. A approach consisted identification active compounds complex plant extract mixtures combination methionine schemes In latter case flavonoids identified VEGF harvested from allowed us mode type molecules. Lastly, much more classic context, decided attempt medium sized oligopeptide NMR. focused Kahalalide F marine origin depsipeptide very promising anticancer activity; although SAR profile would be useful medicinal chemistry perspective molecules still pose challenge their structural particularly stemming intrinsic flexibility. Several media explored including H2O, DMSO, membrane mimics (SDS micelles); peptide seems adopt preferred structure allowing model monomeric insertion within membrane-like environments. Such supports part peptides cytotoxic activity occurs at level is linked formation turn N-terminus section. " RESUMEN: La resonancia magnetica nuclear se utiliza en la actualidad practicamente todas las etapas del proceso de descubrimiento y caracterizacion farmacos. Durante presente tesis nos propusimos explorar estas facetas RMN tomando como modelo diferentes sistemas relacionados con cancer. En primer lugar utilizamos proteinas involucradas para implementar varias metodologias fenomenos interaccion intermolecular. Se pusieron punto esquemas marcaje selectivo triptofano evaluo su aplicabilidad procesos cribado compuestos. Otros experimentos masivo (STD, WaterLOGSY) emplearon identificar nuevos ligandos Bcl-XL; posteriormente, estrategias reconstruccion fragmentos permitieron disenar duales. Las mismas herramientas utilizaron el diseno desarrollo moleculas antiangiogenicas, inhibidoras entre sus receptores. Para este fin, plantearon dos identificacion inhibidores: pretendia un ligando peptidico afinidad elevada partiendo dipeptidos menor utilizando fragmentos. Una segunda estrategia consistia compuestos activos presentes extractos plantas VEGF, concreto metioninas. En ultimo caso consiguieron varios flavonoides capaces interaccionar modo union caracterizo informacion derivada RMN. Por llevo cabo estructural Kahalalido mediante RMN. Este depsipeptido origen marino encuentra fases clinicas contra tipos cancer, presenta enorme interes comprension accion. exploraron preferencias conformacionales peptido medios incluyendo: mimeticos membrana (micelas SDS). El adopta una estructura preferente presencia micelas SDS, lo que permiten proponer insercion monomerica entornos lipidica; esto sugiere parte actividad citotoxica ocurre nivel cual formacion giro ? seccion N-terminal tambien parece relevante. "