Loxapine

Clozapine (catalog number 0444) stock: 50 mg/ml, Haloperidol (catalog number 0931) stock: 10 mg/ml, and Olanzapine (catalog number 4349) stock: 50 mg/ml were from Tocris Bioscience and dissolved in DMSO. Loxapine (catalog number L106) stock: 10 mg/ml was from Sigma-Aldrich and dissolved in 0.9% saline. All aqueous solutions were buffered to pH 6–6.5 if required. The drugs were administered intraperitoneally.
4-OH-Tamoxifen (catalog number H6278) was purchased from Sigma-Aldrich. It was prepared as described previously (Guenthner et al., 2013 (link)). Briefly, 4-OH-Tamoxifen was dissolved in ethanol at 20 mg/ml and stored at -20°C. 4-OH-Tamoxifen stock in ethanol was mixed with corn oil to achieve the final concentration of 10 mg/ml. The ethanol was evaporated using Centrivap before injection.

Mice between the ages of P30 and P40 received an intraperitoneal injection with the antipsychotic drug loxapine (100 mg/kg; Merck Sigma) dissolved in 0.9% sterile saline solution (Pfizer) and 25% DMSO (Merck Sigma). The animals were then monitored for a maximum of 1 hour, and the latency to a first tonic-clonic seizure was recorded.
In the 3 assays, animals were euthanized at the end of the experiment.

Loxapine, pregabalin, chloroquine, SLIGRL‐NH₂, β‐alanine, histamine, PGE₂, serotonin, and bradykinin were purchased from Sigma–Aldrich (Munich, Germany). All indicated concentrations refer to those of pure substances. Reagents and solvents for the synthesis of Loxapine derivatives were obtained from Acros Organics (Gel, Belgium), Alfa Aesar GmbH & Co KG (Karlsruhe, Germany), BLDPharm Inc. (NuiNan, China), Fluorochem Ltd. (Hadfield, UK), Sigma–Aldrich, and TCI Europe N.V. (Zwijndrecht, Belgium).

The main chemicals used in this study, including FGAs (chlorpromazine, chlorprothixene, clothiapine, droperidol, flupentixol, fluphenazine, haloperidol, levomepromazine, loxapine, pimozide, prochlorperazine, sulpiride, thioridazine, and trifluoperazine) and SGAs (amisulpride, aripiprazole, brexpiprazole, clozapine, lurasidone, olanzapine, paliperidone, quetiapine, risperidone, ziprasidone, and zotepine) were acquired from Sigma-Aldrich (St. Louis, Missouri, USA) and were of highest purity grade. Before usage, the compounds were dissolved in dimethyl sulfoxide (DMSO) at the proper concentrations. Human hepatoma HepaRG™ cells were purchased from Thermo Fisher Scientific (Waltham, Massachusetts, USA). For 2 weeks, frozen cells were defrosted and kept alive in Williams’ E medium (Sigma-Aldrich, St. Louis, Missouri, USA) with 10% FetalClone™ II serum (HycloneTM, GE Healthcare, Chicago, Illinois, USA), 1 × L-glutamine, 5 μg/mL human insulin, and 50 μM hydrocortisone hemisuccinate without antibiotics as supplements. To produce differentiated hepatocyte-like features, the medium was then changed to the aforementioned medium plus 2% DMSO for an additional 2 weeks. Cells were grown at 37°C in a humidified 5% CO₂ environment. P-nitrophenylphosphate (PNPP) was used in an acid phosphatase (ACP) experiment to measure cell viability (29 (link)).