Vol. 302, Issue 1, 163-173, July 2002

A Novel N-Methyl-D-aspartate Receptor Open Channel Blocker with in Vivo Neuroprotectant Activity

Rosa Planells-Cases, Carmina Montoliu, Marc Humet, Asia M. Fernández, Carolina García-Martínez, Elvira Valera, Jaime M. Merino, Enrique Pérez-Payá, Angel Messeguer, Vicente Felipo and Antonio Ferrer-Montiel

Centro de Biología Molecular y Celular, Universidad Miguel Hernández, Alicante, Spain (R.P.-C., A.M.F., C.G.M., A.F.-M.); Laboratory of Neurobiology, Instituto de Investigaciones Citológicas, Fundación Valenciana de Investgaciones Biomédicas, Valencia, Spain (C.M., V.F.); Institut d'Investigacions Químiques i Ambientals de Barcelona, Consejo Superior de Investigaciones Cientificas, Barcelona, Spain (M.H., A.M.); Departamento de Bioquímica y Biología Molecular, Universidad de Extremadura, Badajoz, Spain (E.V., J.M.M.); and Departamento de Bioquímica y Biología Molecular, Universidad de Valencia, Valencia, Spain (E.P.-P.)

Excitotoxicity has been implicated in the etiology of ischemic stroke, chronic neurodegenerative disorders, and very recently, in glioma growth. Thus, the development of novel neuroprotectant molecules that reduce excitotoxic brain damage is vigorously pursued. We have used an ionic current block-based cellular assay to screen a synthetic combinatorial library of trimers of N-alkylglycines on the N-methyl-D-aspartate (NMDA) receptor, a well known molecular target involved in excitotoxicity. We report the identification of a family of N-alkylglycines that selectively blocked the NMDA receptor. Notably, compound 3,3-diphenylpropyl-N-glycinamide (referred to as N20C) inhibited NMDA receptor channel activity with micromolar affinity, fast on-off blockade kinetics, and strong voltage dependence. Molecule N20C did not act as a competitive glutamate or glycine antagonist. In contrast, saturation of the blocker binding site with N20C prevented dizolcipine (MK-801) blockade of the NMDA receptor, implying that both drugs bind to the same receptor site. The N-alkylglycine efficiently prevented in vitro excitotoxic neurodegeneration of cerebellar and hippocampal neurons in culture. Attenuation of neuronal glutamate/NMDA-induced Ca²⁺ overload and subsequent modulation of the glutamate-nitric oxide-cGMP pathway seems to underlie N20C neuroprotection. Noteworthy, this molecule exhibited significant in vivo neuroprotectant activity against an acute, severe, excitotoxic insult. Taken together, these findings indicate that N-alkylglycine N20C is a novel, low molecular weight, moderate-affinity NMDA receptor open channel blocker with in vitro and in vivo neuroprotective activity, which, in due turn, may become a tolerated drug for the treatment of neurodegenerative diseases and cancer.