Mechanism of inhibition of the GluR2 AMPA receptor channel opening by 2,3-benzodiazepine derivatives.

2,3-Benzodiazepine derivatives are drug candidates synthesized for potential treatment of various neurodegenerative diseases involving the excessive activity of AMPA receptors. Here we describe a rapid kinetic investigation of the mechanism of inhibition of the GluR2Qflip AMPA receptor channel opening by two 2,3-benzodiazepine derivatives that are structurally similar (BDZ-2 and BDZ-3). Using a laser-pulse photolysis technique with a time resolution of approximately 60 mus, we measured the effects of these inhibitors on both the channel opening rate and the whole-cell current amplitude. We found that both compounds preferably inhibit the open-channel state, although BDZ-2 is a more potent inhibitor in that it inhibits the open-channel state approximately 5-fold stronger than BDZ-3 does. Both compounds bind to the same noncompetitive site. Binding of an inhibitor to the receptor involves the formation of a loose, partially conducting channel intermediate, which rapidly isomerizes to a tighter complex. The isomerization reaction is identified as the main step at which the receptor distinguishes the structural difference between the two compounds. These results suggest that addition of a bulky group at the N-3 position on the diazepine ring, as in BDZ-3, does not alter the mechanism of action, or the site of binding, but does lower the inhibitory potency, possibly due to an unfavorable interaction of a bulky group at the N-3 position with the receptor site. The new mechanistic revelation about the structure-reactivity relationship is useful in designing conformation-specific, more potent noncompetitive inhibitors for the GluR2 AMPA receptor.