Molecular pharmacology of the AMPA agonist, ( S)-2-amino-3-(3-hydroxy-5-phenyl-4-isoxazolyl)propionic acid [( S)-APPA] and the AMPA antagonist, ( R)-APPA

Abstract

The heterocyclic analogue of ( S)-glutamic acid, ( S)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)propionic acid [( S)-AMPA] is a potent and selective AMPA receptor agonist, whereas the enantiomeric compound, ( R)-AMPA, is virtually inactive. We have previously characterized ( RS)-2-amino-3-(3-hydroxy-5-phenyl-4-isoxazolyl)propionic acid [( RS)-APPA] as a partial AMPA receptor agonist showing about 60% of the efficacy of ( RS)-AMPA. This partial agonism produced by ( RS)-APPA is, however, only apparent, since resolution of ( RS)-APPA has now been shown to provide the full AMPA receptor agonist, ( S)-APPA, whereas ( R)-APPA is a non-N- methyl- d-aspartic acid (non-NMDA) receptor antagonist showing preferential AMPA blocking effects. In agreement with classical theories for competitive interaction between agonists and antagonists, the efficacy of depolarizations produced by ( S)-APPA in the rat cortical wedge preparation was shown to be progressively reduced with increasing molar ratios of ( R)-APPA/( S)-APPA. These compounds and the competitive antagonists ( RS)-2-amino-3-(3-carboxymethoxy-5-methyl-4-isoxazolyl)propionic acid [( RS)-AMOA], 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX) and 6-nitro-7-sulfamoylbenzo( f)quinoxalin-2,3-dione (NBQX) were also tested in [ 3H]AMPA and [ 3H]CNQX binding systems, the latter ligand being used in the absence or presence of thiocyanate ions. On the basis of these studies it is suggested that ( RS)-AMPA and the AMPA agonist ( S)-APPA interact with a high-affinity receptor conformation, whereas the competitive antagonists ( RS)-AMOA and ( R)-APPA, derived from these agonists, preferentially bind to a low-affinity AMPA receptor conformation. The competitive antagonists, CNQX and NBQX which are structurally unrelated to ( RS)-AMPA or ( RS)-APPA, do not seem to discriminate between these two AMPA receptor conformations. The modified [ 3H]CNQX binding assay containing thiocyanate ions was shown to provide receptor affinity data for AMPA receptor agonists as well as antagonists, which correlate with the potencies of these compounds in the cortical wedge preparation. Using autoradiographic techniques, ( S)- and ( R)-APPA were shown to exhibit significantly different absolute potencies as inhibitors of [ 3H]AMPA binding in a number of regions of the rat brain.