Characterization of the binding of two novel glycine site antagonists to cloned NMDA receptors: evidence for two pharmacological classes of antagonists

The potency of two novel glycine site antagonists, GV150,526A and GV196,771A, was assessed by their ability to inhibit the binding of [ 3 H]‐MDL105,519 to cell homogenates prepared from mammalian cells transfected with either NR1‐1a, NR1‐2a, NR1‐1a/NR2A, NR1‐1a/NR2B, NR1‐1a/NR2C or NR1‐1a/NR2D NMDA receptor clones. The inhibition constants ( K i s) for GV150,526A displacement of [ 3 H]‐MDL105,519 binding to either NR1‐1a or NR1‐2a expressed alone were not significantly different and were best fit by a one‐site binding model. GV150,526A inhibition to NR1‐1a/NR2 combinations was best fit by a two‐site model with the NR1‐1a/NR2C having an approximate 2–4 fold lower affinity compared to other NR1‐1a/NR2 receptors. The K i s for GV196,771A displacement of [ 3 H]‐MDL105,519 binding to NR1‐1a, NR1‐2a and all NR1‐1a/NR2 combinations was best fit by a two‐site binding model. There was no significant difference between the K i s for the binding to NR1‐1a and NR1‐2a; NR1‐1a/NR2A receptors had an approximate 4 fold lower affinity for GV196,771A compared to other NR1‐1a/NR2 combinations. The K i s for both GV150,526A and GV196,771A for the inhibition of [ 3 H]‐MDL105,519 binding to membranes prepared from adult rat forebrain were determined and compared to the values obtained for binding to cloned NMDA receptors. The K i s for a series of glycine site ligands with diverse chemical structures were also determined for the inhibition of [ 3 H]‐MDL105,519 binding to NR1‐1a/NR2A receptors. L689,560 displayed similar binding characteristics to GV150,526A. It is suggested that glycine site antagonists may be divided into two classes based on their ability to distinguish between NR1 and NR1/NR2 receptors with respect to binding curve characteristics.British Journal of Pharmacology (2000) 130 , 65–72; doi: 10.1038/sj.bjp.0703298