Combination of haloperidol with UNC9994, β-arrestin-biased analog of aripiprazole, ameliorates schizophrenia-related phenotypes induced by NMDAR deficit in mice.

Abstract

Glutamatergic system dysfunction contributes to a full spectrum of schizophrenia-like symptoms, including the cognitive and negative symptoms that are resistant to treatment with antipsychotic drugs (APDs). Aripiprazole, an atypical antipsychotic drug (APD), acts as a dopamine partial agonist and its combination with haloperidol (a typical APD) was suggested as a potential strategy to improve schizophrenia. Recently, an analog of aripiprazole - UNC9994 was developed. UNC9994 does not affect D2R-mediated Gi/o protein signaling but acts as a partial agonist for D2R/β-arrestin interactions. Hence, one of our objectives was to probe the behavioral effects of co-administrating haloperidol with UNC9994 in NMDAR mouse models of schizophrenia. The biochemical mechanisms underlying neurobiological effects of dual haloperidol x UNC9994 action are currently missing. Hence, we aimed to explore D2R- and NMDAR-dependent signaling mechanisms that could underlie the effects of dual drug treatments. NMDAR hypofunction was induced pharmacologically by acute injection of MK-801 (NMDAR pore blocker; 0.15mg/kg) and genetically by knockdown of Grin1 gene expression in mice, which have a 90% reduction in NMDAR levels (Grin1-KD). After intraperitoneal injections of vehicle, haloperidol (0.15mg/kg), UNC9994 (0.25mg/kg) or their combination mice were tested in open field, Pre-Pulse inhibition (PPI), Y-maze and Puzzle box. Biochemical effects on the phosphorylation of Akt, GSK-3 and CaMKII in the prefrontal cortex (PFC) and striatum of MK-801-treated mice were assessed by western blotting. Our findings indicate that low dose co-administration of UNC9994 and haloperidol reduces hyperactivity in MK-801-treated animals and in Grin1-KD mice. Furthermore, this dual administration effectively reverses PPI deficits, repetitive/rigid behavior in the Y-maze, and deficient executive function in the Puzzle box in both animal models. Pharmacological inhibition of NMDAR by MK-801 induced the opposite effects in the PFC and striatum on pAkt-S473 and pGSK3β-Ser9. Dual injection of haloperidol with UNC9994 reversed MK-801-induced effects on pAkt-S473 but not on pGSK3β-Ser9 in both brain structures. The dual administration of haloperidol with UNC9994 at low doses represents a promising approach to ameliorate symptoms of schizophrenia. The combined drug regimen elicits synergistic effects specifically on pAkt-S473, suggesting it as a potential biomarker for antipsychotic' actions.