Tricyclic Antidepressant Structure-Related Alterations in Calcium-Dependent Inhibition and Open-Channel Block of NMDA Receptors

Abstract

N-methyl-D-aspartate receptors (NMDARs) are an essential target for the analgetic action of tricyclic antidepressants (TCAs). Their therapeutic blood concentrations achieve 0.5-1.5 μM, which, however, are insufficient to cause in vitro the open-channel block known as the only effect of TCAs on NMDARs. Whereas structures of amitriptyline (ATL), desipramine (DES), and clomipramine (CLO) are rather similar these compounds manifest different therapeutic profiles and side effects. To study structure-activity relationships of DES and CLO on NMDARs, we measured IC₅₀s as a function of extracellular calcium ([Ca²⁺]) and membrane voltage (V_m) of NMDAR currents recorded in cortical neurons. Here two components of TCA action on NMDARs are described, which could be characterized as the Ca²⁺-dependent inhibition and the open-channel block. DES demonstrated a profound Ca²⁺-dependent inhibition of NMDARs, while the CLO effect was weak. DES IC₅₀ exhibited an e-fold change with a [Ca²⁺] shift of 0.59 mM, which is consistent with ATL. The Ca²⁺ dependence of NMDAR inhibition by DES disappeared in BAPTA loaded neurons, suggesting that Ca²⁺ acts from the inside. Since CLO differs from DES and ATL by the presence of Cl-atom in the structure, most likely, this is the atom which is responsible for the loss of pronounced [Ca²⁺] dependence. As for the NMDAR open-channel block, both DES and CLO were about 5-folds more potent than ATL due to their slow rates of dissociation either from open and closed states. DES demonstrated stronger V_m-dependence than CLO, suggesting a deeper location of the DES binding site within the ion pore. Because DES and CLO differ from ATL by the nitrogen-containing tricycle, presumably this moiety of the molecules determines their high-affinity binding with the NMDAR channel, while the aliphatic chain mono-methyl amino-group of DES allows a deep permeation in the channel. Thus, different structure-activity relationships of the Ca²⁺-dependent inhibition and V_m-dependent open-channel block of NMDARs by DES and CLO suggest that these processes are independent and most likely may represent an action on different molecular targets. The proposed model of TCA action on NMDARs predicts well the experimental values of IC₅₀s at physiological [Ca²⁺] and within a wide range of V_ms.

Keywords: NMDA receptor; amitriptyline; calcium; clomipramine; desipramine; ion channel; tricyclic antidepressants.