Inhibidor altamente selectivo y reversible identificado por la quimioproteómica comparativa Modula la actividad de la diacilglicerol lipasa en las neuronas
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Marc P Baggelaar1, Pascal J P Chameau2, Vasudev Kantae3
1†Department of Bioorganic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden 2300 RA, The Netherlands.
Journal of the American Chemical Society
|June 18, 2015
Resumen
Los investigadores desarrollaron un nuevo inhibidor selectivo, LEI105, para las enzimas diacilglicerol lipasa (DAGL) -α y -β. Este descubrimiento ayuda en el estudio de la señalización endocannabinoide y proporciona evidencia de 2-AG.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- La neurociencia es la neurociencia.
- Farmacología Farmacología.
Sus antecedentes:
- La diacilglicerol lipasa (DAGL) -α y -β son enzimas clave en la síntesis del 2-arachidonoylglycerol (2-AG), un endocannabinoide crucial.
- Se necesitan inhibidores selectivos y reversibles para investigar la función de DAGL en las células neuronales.
- Los inhibidores actuales carecen de la selectividad y el control temporal requeridos para los estudios agudos.
Objetivo del estudio:
- Identificar y caracterizar un inhibidor altamente selectivo para DAGL-α y DAGL-β.
- Para utilizar el diseño guiado por la estructura y la quimioproteómica para el descubrimiento de inhibidores y perfiles de selectividad.
- Investigar el papel del 2-AG en la señalización retrógrada a través de la inhibición de DAGL.
Principales métodos:
- Empleó una estrategia de quimioproteomía que combina el perfil de proteínas basado en la actividad (ABPP) comparativo y competitivo.
- Utilizó fluorofosfonato de amplio espectro y sondas específicas de β-lactona para identificar inhibidores en proteomas complejos.
- Selectividad de inhibidor validada contra otras enzimas metabolizadoras de endocannabinoides y receptores cannabinoides.
Principales resultados:
- Descubrió LEI105, un α-cetoheterociclo, como un potente, selectivo y reversible doble inhibidor de DAGL-α/DAGL-β.
- LEI105 no demostró inhibición de las enzimas relacionadas (ABHD6, ABHD12, MAGL, FAAH) o afinidad por los receptores CB1.
- LEI105 redujo los niveles de 2-AG en las células Neuro2A e inhibió la plasticidad sináptica mediada por el receptor CB1 en rebanadas hipocampales de ratón.
Conclusiones:
- LEI105 es una valiosa herramienta farmacológica para estudiar la función de DAGL y la señalización endocannabinoide.
- Los hallazgos apoyan la hipótesis de la biosíntesis de 2-AG "bajo demanda" en la señalización retrógrada.
- Este estudio pone de relieve el poder de ABPP en el descubrimiento de inhibidores selectivos de enzimas dentro de los sistemas biológicos nativos.
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