BETMB: A Dual-Target Compound for Synergistic Suppression of Neuronal Hyperexcitability in Refractory Epilepsy via Concurrent Modulation of Nav Channels and GABAA Receptors.

AIMS: This study aims to evaluate 5-(but-1-en-1-yl)-1,2,3-trimethoxybenzene (BETMB) as a novel dual-target anti-seizure agent for refractory epilepsy and elucidate the synergistic neuroelectrophysiological mechanism between NaV channels and GABAA receptors. METHODS: Whole-cell patch-clamp recordings characterized BETMB's dual-target activity. Antiseizure efficacy was assessed in maximal electroshock (MES), pentylenetetrazole (PTZ), and kainic acid (KA) models. Cognitive function in chronic KA mice was evaluated using the Morris water maze (MWM). Histopathological, immunohistochemical, and Western blot analyses explored neuroprotection. Synergy between NaV and GABAAR modulation was systematically investigated using both an in vitro Mg2+-free model of neuronal hyperexcitability and an in silico model of cortical spreading depolarization (CSD). RESULTS: BETMB acted as a GABAAR positive allosteric modulator (EC50 = 93.2 μM) and a state-dependent NaV blocker (KI = 1.9 μM). It significantly suppressed seizures across models, improved cognition in chronic epilepsy, and modulated downstream expression of GABRA1, NR2B, and BDNF-pAKT-CREB signaling. Synergistic NaV and GABAAR modulation completely abolished ictal-like discharges in Mg2+-free cellular models and prevented CSD initiation in computational simulations. CONCLUSION: BETMB is a promising dual-target therapy for refractory epilepsy, supported by the first electrophysiological evidence that dual modulation of GABAAR and NaV synergistically suppresses neuronal hyperexcitability. Beyond epilepsy, this finding may also extend to CSD-related conditions such as stroke, traumatic brain injury, and migraine.