Functional in vitro characterization of CR 3394: A novel voltage dependent N-methyl- d-aspartate (NMDA) receptor antagonist

• Published in: Neuropharmacology Vol. 50; no. 3; pp. 277 - 285
• Main Authors: Losi, Gabriele, Lanza, Marco, Makovec, Francesco, Artusi, Roberto, Caselli, Gianfranco, Puia, Giulia
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.03.2006
• Subjects: Adamantane - analogs & derivatives; Adamantane - pharmacology; Amidines - pharmacology; Animals; Animals, Newborn; Cells, Cultured; Cerebral Cortex - cytology; Dopamine - metabolism; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Drug Interactions; Electric Stimulation - methods; Electrophysiology; Excitatory Amino Acid Antagonists - pharmacology; Humans; In Vitro Techniques; Membrane Potentials - drug effects; Membrane Potentials - physiology; Membrane Potentials - radiation effects; N-Methylaspartate - pharmacology; Neural Inhibition - drug effects; Neurodegenerative disorders; Neurons - drug effects; NMDA antagonists; NMDA receptors; Norepinephrine - metabolism; Patch-clamp; Patch-Clamp Techniques - methods; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate - antagonists & inhibitors; Receptors, N-Methyl-D-Aspartate - genetics; Receptors, N-Methyl-D-Aspartate - metabolism; Transfection - methods; Tritium - metabolism; NMDA antagonists; Electrophysiology; NMDA receptors; Patch-clamp; Neurodegenerative disorders
• ISSN: 0028-3908; 1873-7064
• DOI: 10.1016/j.neuropharm.2005.09.002

Using the patch-clamp technique, we studied the effect of two novel adamantane derivatives, N-[2-(3,5-dimethyl-1-adamantyl)ethyl] guanidine (CR 3391) and N-[2-(3,5-dimethyl-1-adamantyl) ethyl]acetamidine (CR 3394), on NMDA receptors expressed in cortical neuron cultures. Our data show that CR 3391 and CR 3394 reduce NMDA-evoked currents (IC 50 = 1.7 ± 0.6 μM and 6.7 ± 1.5 μM, respectively). This antagonism is non-competitive and is completely reversible. The effect of CR 3394, like that of memantine, was strongly voltage dependent. HEK293 cells expressing NR1a/NR2B recombinant NMDA receptors and immature neurons (DIV 8–9) were more sensitive to CR 3394 antagonism than NR1a/NR2A expressing cells and DIV 15 neurons. CR 3394 also reduced the duration and amplitude of miniature excitatory post-synaptic currents mediated exclusively by NMDA receptors (NMDA-mEPSCs). Both memantine and CR 3394 inhibited NMDA-evoked [ 3H]norepinephrine release from rat hippocampal slices in a concentration-dependent manner with similar potency. CR 3394, but not memantine, increased cathecholamine resting release at low micromolar concentrations. Moreover, in an in vitro model of neurotoxicity, CR 3394 strongly reduced glutamate- and NMDA-induced neuronal death. Taken together, our data highlight pharmacological features of CR 3394 in vitro that prompt us to further evaluate it as a candidate for the treatment of neurodegenerative disorders.