Two subtypes of
cannabinoid receptors are currently recognized, CB(1), found in brain and neuronal cells, and CB(2), found in spleen and immune cells. We have characterized 1-(2-chlorophenyl)-4-cyano-5-(4-methoxyphenyl)-1H-pyrazole-3-carboxyl ic
acid phenylamide (CP-272871) as a novel aryl
pyrazole antagonist for the CB(1) receptor.
CP-272871 competed for binding of the
cannabinoid agonist (3)H-labeled (-)-3-[2-hydroxy-4-(1, 1-dimethylheptyl)-phenyl]-4-[3-hydroxypropyl]cyclohexan-1-ol ([(3)H]
CP-55940) at the CB(1) receptor in rat brain membranes with a K(d) value 20-fold greater than that of N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide HCl (
SR141716A).
CP-272871 also competed for binding with the aminoalkylindole agonist (3)H-labeled (R)-(+)-[2, 3-dihydro-5-methyl-3-[(4-morpholinyl)methyl]pyrrolo[1,2,3-de]1, 4-benzoxazin-6-yl](1-naphthyl)methanone ([(3)H]
WIN-55212-2), as well as the aryl
pyrazole antagonist [(3)H]
SR141716A. Inverse agonist as well as antagonist properties were observed for both
SR141716A and
CP-272871 in signal transduction assays in
biological preparations in which the CB(1) receptor is endogenously expressed.
SR141716A augmented
secretin-stimulated
cyclic AMP (cAMP) accumulation in intact N18TG2
neuroblastoma cells, and this response was reversed by the agonist
desacetyllevonantradol.
CP-272871 antagonized
desacetyllevonantradol-mediated inhibition of
adenylyl cyclase in N18TG2 membranes, and increased
adenylyl cyclase activity in the absence of agonist.
SR141716A and
CP-272871 antagonized
desacetyllevonantradol-stimulated (35)S-labeled guanosine-5'-O-(gamma-thio)-triphosphate ([(35)S]
GTPgammaS) binding to brain membrane
G-proteins, and decreased basal [(35)S]
GTPgammaS binding to
G-proteins. K(+) enhanced
CP-272871 and
SR141716A inverse agonist activity compared with Na(+) or NMDG(+) in the assay. These results demonstrated that the aryl
pyrazoles SR141716A and
CP-272871 behave as antagonists and as inverse agonists in
G-protein-mediated signal transduction in preparations of endogenously expressed CB(1) receptors.