Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors
Excessive Ca<sup>2+</sup> currents via N-methyl-D-aspartate receptors (NMDARs) have been implicated in many disorders. Uncompetitive NMDAR channel blockers are an emerging class of drugs in clinical use for major depressive disorder (MDD) and other neuropsychiatric diseases. The pharmaco...
| Main Authors: | , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-08-01
|
| Series: | Pharmaceuticals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1424-8247/15/8/997 |
| _version_ | 1797408454398181376 |
|---|---|
| author | Ezio Bettini Stephen M. Stahl Sara De Martin Andrea Mattarei Jacopo Sgrignani Corrado Carignani Selena Nola Patrizia Locatelli Marco Pappagallo Charles E. Inturrisi Francesco Bifari Andrea Cavalli Andrea Alimonti Luca Pani Maurizio Fava Sergio Traversa Franco Folli Paolo L. Manfredi |
| author_facet | Ezio Bettini Stephen M. Stahl Sara De Martin Andrea Mattarei Jacopo Sgrignani Corrado Carignani Selena Nola Patrizia Locatelli Marco Pappagallo Charles E. Inturrisi Francesco Bifari Andrea Cavalli Andrea Alimonti Luca Pani Maurizio Fava Sergio Traversa Franco Folli Paolo L. Manfredi |
| author_sort | Ezio Bettini |
| collection | DOAJ |
| description | Excessive Ca<sup>2+</sup> currents via N-methyl-D-aspartate receptors (NMDARs) have been implicated in many disorders. Uncompetitive NMDAR channel blockers are an emerging class of drugs in clinical use for major depressive disorder (MDD) and other neuropsychiatric diseases. The pharmacological characterization of uncompetitive NMDAR blockers in clinical use may improve our understanding of NMDAR function in physiology and pathology. REL-1017 (esmethadone-HCl), a novel uncompetitive NMDAR channel blocker in Phase 3 trials for the treatment of MDD, was characterized together with dextromethorphan, memantine, (±)-ketamine, and MK-801 in cell lines over-expressing NMDAR subtypes using fluorometric imaging plate reader (FLIPR), automated patch-clamp, and manual patch-clamp electrophysiology. In the absence of Mg<sup>2+</sup>, NMDAR subtypes NR1-2D were most sensitive to low, sub-μM glutamate concentrations in FLIPR experiments. FLIPR Ca<sup>2+</sup> determination demonstrated low μM affinity of REL-1017 at NMDARs with minimal subtype preference. In automated and manual patch-clamp electrophysiological experiments, REL-1017 exhibited preference for the NR1-2D NMDAR subtype in the presence of 1 mM Mg<sup>2+</sup> and 1 μM L-glutamate. Tau off and trapping characteristics were similar for (±)-ketamine and REL-1017. Results of radioligand binding assays in rat cortical neurons correlated with the estimated affinities obtained in FLIPR assays and in automated and manual patch-clamp assays. In silico studies of NMDARs in closed and open conformation indicate that REL-1017 has a higher preference for docking and undocking the open-channel conformation compared to ketamine. In conclusion, the pharmacological characteristics of REL-1017 at NMDARs, including relatively low affinity at the NMDAR, NR1-2D subtype preference in the presence of 1 mM Mg<sup>2+</sup>, tau off and degree of trapping similar to (±)-ketamine, and preferential docking and undocking of the open NMDAR, could all be important variables for understanding the rapid-onset antidepressant effects of REL-1017 without psychotomimetic side effects. |
| first_indexed | 2024-03-09T03:58:45Z |
| format | Article |
| id | doaj.art-a9489daf59de4062bfa561e5de1336f9 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-09T03:58:45Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-a9489daf59de4062bfa561e5de1336f92023-12-03T14:16:28ZengMDPI AGPharmaceuticals1424-82472022-08-0115899710.3390/ph15080997Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate ReceptorsEzio Bettini0Stephen M. Stahl1Sara De Martin2Andrea Mattarei3Jacopo Sgrignani4Corrado Carignani5Selena Nola6Patrizia Locatelli7Marco Pappagallo8Charles E. Inturrisi9Francesco Bifari10Andrea Cavalli11Andrea Alimonti12Luca Pani13Maurizio Fava14Sergio Traversa15Franco Folli16Paolo L. Manfredi17In Vitro Pharmacology Department, Aptuit, An Evotec Company, 37135 Verona, ItalyDepartment of Psychiatry, VAMC (SD), University of California, San Diego, CA 92093, USADepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35122 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35122 Padua, ItalyInstitute for Research in Biomedicine (IRB), Università della Svizzera Italiana (USI), 6500 Bellinzona, SwitzerlandIn Vitro Pharmacology Department, Aptuit, An Evotec Company, 37135 Verona, ItalyIn Vitro Pharmacology Department, Aptuit, An Evotec Company, 37135 Verona, ItalyInstitute for Research in Biomedicine (IRB), Università della Svizzera Italiana (USI), 6500 Bellinzona, SwitzerlandDepartment of Anesthesiology, Albert Einstein College of Medicine, Bronx, NY 10461, USARelmada Therapeutics, Coral Gables, FL 33134, USADepartment of Medical Biotechnology and Translational Medicine, University of Milan, 20122 Milan, ItalyInstitute for Research in Biomedicine (IRB), Università della Svizzera Italiana (USI), 6500 Bellinzona, SwitzerlandDepartment of Health Sciences and Technology, ETH Zurich, 8092 Zurich, SwitzerlandRelmada Therapeutics, Coral Gables, FL 33134, USADepartment of Psychiatry, Massachusetts General Hospital, Boston, MA 02114, USARelmada Therapeutics, Coral Gables, FL 33134, USADepartment of Health Sciences, University of Milan, 20122 Milan, ItalyRelmada Therapeutics, Coral Gables, FL 33134, USAExcessive Ca<sup>2+</sup> currents via N-methyl-D-aspartate receptors (NMDARs) have been implicated in many disorders. Uncompetitive NMDAR channel blockers are an emerging class of drugs in clinical use for major depressive disorder (MDD) and other neuropsychiatric diseases. The pharmacological characterization of uncompetitive NMDAR blockers in clinical use may improve our understanding of NMDAR function in physiology and pathology. REL-1017 (esmethadone-HCl), a novel uncompetitive NMDAR channel blocker in Phase 3 trials for the treatment of MDD, was characterized together with dextromethorphan, memantine, (±)-ketamine, and MK-801 in cell lines over-expressing NMDAR subtypes using fluorometric imaging plate reader (FLIPR), automated patch-clamp, and manual patch-clamp electrophysiology. In the absence of Mg<sup>2+</sup>, NMDAR subtypes NR1-2D were most sensitive to low, sub-μM glutamate concentrations in FLIPR experiments. FLIPR Ca<sup>2+</sup> determination demonstrated low μM affinity of REL-1017 at NMDARs with minimal subtype preference. In automated and manual patch-clamp electrophysiological experiments, REL-1017 exhibited preference for the NR1-2D NMDAR subtype in the presence of 1 mM Mg<sup>2+</sup> and 1 μM L-glutamate. Tau off and trapping characteristics were similar for (±)-ketamine and REL-1017. Results of radioligand binding assays in rat cortical neurons correlated with the estimated affinities obtained in FLIPR assays and in automated and manual patch-clamp assays. In silico studies of NMDARs in closed and open conformation indicate that REL-1017 has a higher preference for docking and undocking the open-channel conformation compared to ketamine. In conclusion, the pharmacological characteristics of REL-1017 at NMDARs, including relatively low affinity at the NMDAR, NR1-2D subtype preference in the presence of 1 mM Mg<sup>2+</sup>, tau off and degree of trapping similar to (±)-ketamine, and preferential docking and undocking of the open NMDAR, could all be important variables for understanding the rapid-onset antidepressant effects of REL-1017 without psychotomimetic side effects.https://www.mdpi.com/1424-8247/15/8/997REL-1017esmethadone HCLN-methyl-D-aspartate receptor (NMDAR)dextromethadoned-methadoneketamine |
| spellingShingle | Ezio Bettini Stephen M. Stahl Sara De Martin Andrea Mattarei Jacopo Sgrignani Corrado Carignani Selena Nola Patrizia Locatelli Marco Pappagallo Charles E. Inturrisi Francesco Bifari Andrea Cavalli Andrea Alimonti Luca Pani Maurizio Fava Sergio Traversa Franco Folli Paolo L. Manfredi Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors Pharmaceuticals REL-1017 esmethadone HCL N-methyl-D-aspartate receptor (NMDAR) dextromethadone d-methadone ketamine |
| title | Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors |
| title_full | Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors |
| title_fullStr | Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors |
| title_full_unstemmed | Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors |
| title_short | Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors |
| title_sort | pharmacological comparative characterization of rel 1017 esmethadone hcl and other nmdar channel blockers in human heterodimeric n methyl d aspartate receptors |
| topic | REL-1017 esmethadone HCL N-methyl-D-aspartate receptor (NMDAR) dextromethadone d-methadone ketamine |
| url | https://www.mdpi.com/1424-8247/15/8/997 |
| work_keys_str_mv | AT eziobettini pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT stephenmstahl pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT sarademartin pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT andreamattarei pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT jacoposgrignani pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT corradocarignani pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT selenanola pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT patrizialocatelli pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT marcopappagallo pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT charleseinturrisi pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT francescobifari pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT andreacavalli pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT andreaalimonti pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT lucapani pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT mauriziofava pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT sergiotraversa pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT francofolli pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors AT paololmanfredi pharmacologicalcomparativecharacterizationofrel1017esmethadonehclandothernmdarchannelblockersinhumanheterodimericnmethyldaspartatereceptors |