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1Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Journal of the American Chemical Society
|March 10, 2010
Resumen
Desarrollamos nuevos inhibidores bifaciales que controlan la actividad enzimática al unirse tanto a las enzimas como a los cucurbituriles (CB[7]). Esto permite la activación y desactivación reversible de la actividad de la anidrasa carbónica bovina (BCA), a diferencia de la inhibición estable de la acetilcolinesterasa (AChE).
Área de la Ciencia:
- Química supramolecular de las moléculas.
- La inhibición de la enzima inhibición de la enzima.
- Biología Química Biología química.
Sus antecedentes:
- La modulación de la actividad enzimática es crucial para los procesos biológicos y las intervenciones terapéuticas.
- Los Cucurbiturils (CBs) son anfitriones macrocíclicos capaces de formar complejos de inclusión estables con varios huéspedes.
- El desarrollo de inhibidores controlables de enzimas sigue siendo un desafío significativo en la biología química.
Objetivo del estudio:
- Sintetizar y caracterizar nuevos inhibidores "de dos caras" que poseen tanto dominios de unión enzimática como de unión al cucurbiturilo.
- Investigar la inhibición controlable de la anidrasa carbónica bovina (BCA) y la acetilcolinesterasa (AChE) utilizando estos inhibidores y la CB[7].
- Aclarar el mecanismo de la inhibición enzimática reversible e irreversible a través de la formación de complejos supramoleculares.
Principales métodos:
- Síntesis de moléculas bifuncionales con inhibidores de enzimas y sitios de reconocimiento de cucurbiturilo.
- Análisis de actividad enzimática (UV/Vis) para monitorear las tasas catalíticas de BCA y AChE.
- Ensayos de desplazamiento de fluorescencia, espectroscopia de RMN de 1H y mediciones cinéticas (Ka, kon, koff) para caracterizar la formación y la disociación complejas.
Principales resultados:
- Se han sintetizado con éxito inhibidores bifaciales 1-5 dirigidos a BCA y AChE.
- Se ha demostrado la activación y desactivación reversible de la actividad de BCA a través de la formación y disociación de complejos ternales transitorios BCA-CB[7].
- Se logra una inhibición estable e irreversible de la AChE a través de la formación de complejos ternales AChE-CB termodinámicamente estables[7].
Conclusiones:
- La estrategia inhibidora "de dos caras" permite un control sintonizable sobre la actividad catalítica de la enzima.
- La inhibición reversible se logró con BCA, mientras que la inhibición irreversible se observó con AChE, dependiendo de la estabilidad del complejo.
- Este enfoque ofrece un mecanismo distinto para el control de la actividad enzimática en comparación con la regulación alosterica natural.
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