PURPOSE: A thermodynamic analysis of the binding to rat cortex adenosine A1 receptor of N6-substituted (full agonists) and N6-substituted-deoxyribose (partial agonists) adenosine derivatives was performed. The intrinsic activity of the compounds was evaluated by measurements of the inhibition of forskolin stimulated 3', 5'-cyclic adenosine monophosphate (c-AMP) levels in isolated epididymal rat adipocytes. METHODS: The thermodynamic parameters deltaG(o) (standard free energy), deltaH(o) (standard enthalpy), and deltaS(o) (standard entropy) of the binding equilibrium were determined by means of affinity measurements carried out at different temperatures (0, 10, 20, 25, 30 degrees C). Levels of c-AMP were evaluated performing competitive protein binding assays. RESULTS: The binding of the ligands increases with temperature enhancement and, as a consequence, is totally entropy driven. Standard entropy values correlate significantly with intrinsic activity ones. CONCLUSIONS: It is proposed the data obtained by these in vitro experiments can be used to investigate the in vivo pharmacodynamic of A, full and partial agonists.
Thermodynamic in vitro studies as a method to investigate the pharmacodynamic behavior of adenosine A(1) receptor ligands
DALPIAZ, Alessandro;SCATTURIN, Angelo;PAVAN, Barbara;VARANI, Katia;BOREA, Pier Andrea
1999
Abstract
PURPOSE: A thermodynamic analysis of the binding to rat cortex adenosine A1 receptor of N6-substituted (full agonists) and N6-substituted-deoxyribose (partial agonists) adenosine derivatives was performed. The intrinsic activity of the compounds was evaluated by measurements of the inhibition of forskolin stimulated 3', 5'-cyclic adenosine monophosphate (c-AMP) levels in isolated epididymal rat adipocytes. METHODS: The thermodynamic parameters deltaG(o) (standard free energy), deltaH(o) (standard enthalpy), and deltaS(o) (standard entropy) of the binding equilibrium were determined by means of affinity measurements carried out at different temperatures (0, 10, 20, 25, 30 degrees C). Levels of c-AMP were evaluated performing competitive protein binding assays. RESULTS: The binding of the ligands increases with temperature enhancement and, as a consequence, is totally entropy driven. Standard entropy values correlate significantly with intrinsic activity ones. CONCLUSIONS: It is proposed the data obtained by these in vitro experiments can be used to investigate the in vivo pharmacodynamic of A, full and partial agonists.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
