NMDA receptor subtype differential affinity as a key enabler for precision neuropsychiatry

Despite recent advances in neuropsychiatry, translational development for many targets remains challenging, particularly when receptors exist as families with complex subunit-dependent pharmacology and safety concerns. This is the case for NMDA receptor (NMDAR) antagonists. R,S-ketamine illustrates this issue: although it produces rapid antidepressant effects in the clinic, its use is limited by dissociative side effects and potential long-term cognitive impairment. Preclinical studies have also linked it to neuropathological alterations such as Olney's lesions. These limitations highlight the need for deeper mechanistic characterization beyond simple receptor inhibition. Recently, we investigated RST-01, a novel fluoroalkyl derivative of amantadine, as a potential treatment for early post-traumatic stress disorder (PTSD). In preclinical rat models, RST-01 showed efficacy without observable dissociation and without inducing Olney's lesions, an established marker of neurotoxicity unlike R,S-ketamine and memantine reported in the literature. To better understand these safety differences, we examined the target-engagement profiles of RST-01, ketamine, and memantine in preclinical systems. Specifically, we assessed their selectivity across NMDAR subtypes. Using in-vitro electrophysiological assays, we determined IC₅₀ values for human GluN1/GluN2A-D receptors and applied a multidimensional analytical approach to compare their subunit selectivity patterns. Our results revealed distinct pharmacological signatures across the four GluN2 subunits. When represented in multidimensional space, RST-01 displayed a more differentiated selectivity profile than ketamine or memantine. This subunit-specific characterization provides a mechanistic framework to interpret differences in adverse-effect profiles and safety margins observed in preclinical models, supporting the development of safer and more selective NMDAR-targeting therapies.