cgp-39653 and arcaine

This study investigated the binding of [3H]CGP 39653, a novel high-affinity antagonist of the N-methyl-D-aspartate (NMDA) recognition site of the NMDA receptor complex. [3H]CGP 39653 bound to the NMDA receptor in well washed rat brain membranes with an affinity of about 15 nM. Other NMDA site drugs inhibited [3H]CGP 39653 binding with the following order of potency: DL-(tetrazol-5-yl)glycine > glutamate > CGS 19755 > DL-2-amino-5-phosphonovalerate (DL-AP5) > NMDA. Glycine and 5,7-dichlorokynurenate partially inhibited binding. The polyamines spermine and spermidine increased [3H]CGP 39653 binding (EC50 values of 10 and 22 microM, respectively). This effect was mimicked by arcaine, 1,5-diethylaminopiperidine, diaminodecane, diethylenetriamine, and Mg2+. The increase in [3H]CGP 39653 was a result of an increased affinity of the binding site for the ligand with very little effect on binding site density. Spermine and Mg2+ also increased the affinity of the antagonists DL-AP5 and CGS 19755, but had only minor effects on the affinity of glutamate and NMDA. Arcaine did not reverse the enhancement of [3H]CGP 39653 binding by spermine, spermidine, or Mg2+. Channel-blocking dissociative anesthetics, including dizocilpine and ketamine, did not alter basal or Mg(2+)-stimulated [3H]CGP 39653 binding. Spermine did not alter either the enhancement of [3H]dizocilpine by glutamate or the inhibition of [3H]dizocilpine by DL-AP5 or CGS 19755. These studies show that polyamines and divalent cations selectively enhance the affinity of antagonists for the agonist binding site on the NMDA receptor complex. However, this effect is mediated by a site independent of the primary polyamine site defined using [3H]dizocilpine binding.

Topics: 2-Amino-5-phosphonovalerate; Animals; Biguanides; Binding Sites; Binding, Competitive; Brain; Cell Membrane; Glycine; Kinetics; Magnesium; Polyamines; Rats; Receptors, N-Methyl-D-Aspartate; Spermidine; Spermine; Tritium

7-Chlorokynurenate, an antagonist at the glycine recognition site of the NMDA receptor complex, increases the binding of the competitive NMDA receptor antagonist [3H]CGP39653 ([3H]D,L-(E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid) to well washed rat brain membranes but only in the presence of 100 microM spermine. Conversely, spermine only increases [3H]CGP39653 binding in the presence of 10 microM 7-chlorokynurenate, through a mechanism insensitive to the putative polyamine antagonists ifenprodil, arcaine or putrescine. Thus, the effects of glycine antagonists and polyamines on the binding of competitive NMDA receptor antagonists may depend on the residual glycine and polyamine content of the membrane preparation or the state of the glycine recognition site. These data further attest to the complexity of interactions between spermine and the glycine and glutamate recognition sites of the NMDA receptor.

Topics: 2-Amino-5-phosphonovalerate; Allosteric Regulation; Animals; Biguanides; Binding Sites; Glycine; In Vitro Techniques; Kynurenic Acid; Male; Piperidines; Prosencephalon; Putrescine; Radioligand Assay; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Spermine