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. 1991 Sep;104(1):207–221. doi: 10.1111/j.1476-5381.1991.tb12409.x

Structure-activity analysis of binding kinetics for NMDA receptor competitive antagonists: the influence of conformational restriction.

M Benveniste 1, M L Mayer 1
PMCID: PMC1908275  PMID: 1686203

Abstract

1. The kinetics of action of 17 structurally related NMDA receptor competitive antagonists were measured under voltage clamp in mouse hippocampal neurones. Analysis of the response to rapid changes in antagonist concentration during constant application of agonist was used to estimate microscopic association (kon) and dissociation (koff) rate constants for antagonist binding, assuming a two-equivalent site model for competitive antagonism. Dose-inhibition curves were analysed to estimate antagonist equilibrium dissociation constants. 2. For a series of 11 omega-phosphono, alpha-amino acids kon and koff varied 26 and 107 fold respectively. Rapid association and dissociation rate constants were obtained for flexible antagonist molecules such as D-2-amino-7-phosphonoheptanoic acid (D-AP7): kon 1.4 x 10(7) M-1 s-1; koff 20.3 s-1. For conformationally restrained molecules such as 3S,4aR,6S,8aR-6-phosphonomethyl-decahydroisoquinoline- 3-carboxylic acid (LY 235959), association and dissociation rate constants were much slower: kon 1.1 x 10(6) M-1 s-1; koff 0.2 s-1. For the D- and L-isomers of 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) estimates for kon were similar, but for the L-isomer koff was 10 fold faster than for the D-isomer. 3. For 2-amino-5-phosphonopentanoic acid (AP5) and its piperidine derivative cis-4-(phosphonomethyl)piperidine-2-carboxylic acid (CGS 19755), an increase in chain length of two methylene groups between the omega-phosphono and alpha-carboxylate moieties caused a 1.6 to 1.8 fold decrease in kon with little change in koff. In contrast, for AP5, CPP and its omega-carboxylate analogue, addition of a double bond close to the phosphonate moiety caused a 1.3 to 1.6 fold increase in kon. 4. For antagonists with an omega-tetrazole moiety, kon and koff were 2.8-4.6 times faster than for the parent omega-phosphono compounds. A similar, but smaller increase in kon and koff was observed for antagonists with an omega-carboxylate moiety. 5. The slow kinetics of action of potent NMDA receptor antagonists were not an artefact of buffered diffusion. In neurones equilibrated with 200 microM D-AP7, 2 microM LY 235959 and 10 microM NMDA, a transient agonist response was recorded following a rapid switch to D-AP7-free solution. This can only be explained by differences in the binding kinetics of AP7 and LY 235959, since at equilibrium, with these concentrations, either antagonist essentially eliminates the agonist response to 10 microM NMDA.(ABSTRACT TRUNCATED AT 400 WORDS)

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Selected References

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