Binding thermodynamics at the human cannabinoid CB1 and CB2 receptors.
The thermodynamic parameters DeltaG degrees , DeltaH degrees and DeltaS degrees of the binding equilibrium of agonists and antagonists at cannabinoid CB(1) and CB(2) receptors were determined by means of affinity measurements at different temperatures and van't Hoff plots were constructed. Affinity constants were measured on CHO cells transfected with the human CB(1) and CB(2) receptors by inhibition assays of the binding of the cannabinoid receptor agonist [(3)H]-CP-55,940. van't Hoff plots were linear for agonists and antagonists in the temperature range 0-30 degrees C. The thermodynamic parameters for CB(1) receptors fall in the ranges 17< or =DeltaH degrees < or =59 kJ/mol and 213< or =DeltaS degrees < or =361 kJ/mol for agonists and -52< or =DeltaH degrees < or =-26 kJ/mol and -12< or =DeltaS degrees < or =38 kJ/mol for antagonists. The thermodynamic parameters for CB(2) receptors fall in the ranges 27< or =DeltaH degrees < or =48 kJ/mol and 234< or =DeltaS degrees < or =300 kJ/mol for agonists and -19< or =DeltaH degrees < or =-17 kJ/mol and 43< or =DeltaS degrees < or =74 kJ/mol for antagonists. Collectively, these data show that agonist binding is always totally entropy-driven while antagonist binding is enthalpy and entropy-driven, indicating that CB(1) and CB(2) receptors are thermodynamically discriminated. These data could give new details on the nature of the forces driving the CB(1) and CB(2) binding at a molecular level. Enthalpy, entropy, free energy and binding affinity for each ligand to its receptor can all be assessed and therefore the optimal binding profile discovered. Carrying out these binding investigations as early as possible in the discovery process increases the probability that a lead compound will become a successful pharmaceutical compound.
Merighi
Stefania
S
Department of Clinical and Experimental Medicine, Pharmacology Section and Interdisciplinary Center for the Study of Inflammation, University of Ferrara, Via Fossato di Mortara 17/19, 44100 Ferrara, Italy.
Simioni
Carolina
C
Gessi
Stefania
S
Varani
Katia
K
Borea
Pier Andrea
PA
D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects / Q000502:physiology; D016466:CHO Cells; D002186:Cannabinoids / Q000378:metabolism / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D043884:Receptor, Cannabinoid, CB1 / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013816:Thermodynamics
D002186:Cannabinoids; D043884:Receptor, Cannabinoid, CB1; D043885:Receptor, Cannabinoid, CB2