GluN2C selective inhibition is a target to develop new antiepileptic compounds

Abstract Objective Many early‐onset epilepsies present as developmental and epileptic encephalopathy associated with refractory seizures, altered psychomotor development, and disorganized interictal cortical activity. Abnormal upregulation of specific N ‐methyl‐ d ‐aspartate receptor (NMDA‐R) subunits is being disentangled as one of the mechanisms of severe early‐onset epilepsies. In tuberous sclerosis complex (TSC), upregulation of the GluN2C subunit of the NMDA‐R with slow deactivation kinetic results in increased neuronal excitation and synchronization. Methods Starting from an available GluN2C/D antagonist, NMDA‐R–modulating compounds were developed and screened using a patch clamp on neuronal culture to select those with the strongest inhibitory effect on glutamatergic NMDA currents. For these selected compounds, blood pharmacokinetics and passage through the blood–brain barrier were studied. We tested the effect of the most promising compounds on epileptic activity in Tsc1 + /− mice brain slices with multielectrode array, and then in vivo at postnatal ages P14–P17, comparable with the usual age at epilepsy onset in human TSC. Results Using a double‐electrode voltage clamp on isolated NMDA currents, we identified the most prominent antagonists of the GluN2C subunit with no effect on GluN2A as a means of preventing side effects. The best compound passing through the blood–brain barrier was selected. Applied in vivo in six Tsc1 + /− mice at P14–P17, this compound reduced or completely stopped spontaneous seizures in four of them, and decreased the background activity disorganization. Furthermore, ictal‐like discharges stopped on a human brain sample from an infant with epilepsy due to TSC. Interpretation Subunit‐selective inhibition is a valuable target for developing drugs for severe epilepsies resulting from an upregulation of NMDA‐R subunit–mediated transmission.