Deschloroclozapine, a potent and selective chemogenetic actuator enables rapid neuronal and behavioral modulations in mice and monkeys

Abstract

The chemogenetic technology Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) affords remotely reversible control of cellular signaling, neuronal activity and behavior. Although the combination of muscarinic-based DREADDs with clozapine-Noxide (CNO) has been widely used, sluggish kinetics, metabolic liabilities, and potential offtarget effects of CNO represent areas for improvement. Here we provide a new high affinity and selective agonist deschloroclozapine (DCZ) for muscarinic-based DREADDs. Positron emission tomography revealed that DCZ selectively bound to and occupied DREADDs in both mice and monkeys. Systemic delivery of low doses of DCZ (1 or 3 μg/kg) enhanced neuronal activity via hM3Dq within minutes in mice and monkeys. Intramuscular injections of DCZ (100 μg/kg) reversibly induced spatial working memory deficits in monkeys expressing hM4Di in the prefrontal cortex. DCZ represents the most potent, selective, metabolically stable and fast-acting DREADD agonist reported with utility in both mice and non-human primates for a variety of applications.

Effect of chemogenetic actuator drugs on prefrontal cortex-dependent working memory in nonhuman primates

“…Both actuators are FDA-approved which could facilitate their application for chemogenetic technologies in a clinical setting. Deschloroclozapine is a dual dopamine and serotonin receptor antagonist with some selectivity for D 1 ( K i = 200 n m ), 5-HT 2A ( K i = 87 n m ), and M 5 receptors ( K i = 55 n m ) [ 18 ]. A dose of 0.1 mg/kg i.m.…”

“…A dose of 0.1 mg/kg i.m. of deschloroclozapine is effective at activating DREADD receptors in vivo and reversibly inducing behavioral effects in monkeys [ 18 ].…”

“…Three of these new actuators—low-dose clozapine, low-dose olanzapine, and deschloroclozapine—possess better brain penetrance and higher affinity and potency for DREADD receptors than CNO. All of these actuators have been tested in rodent systems [ 8 , 13 , 16 ] and low-dose clozapine [ 17 ] and deschloroclozapine [ 18 ] have been tested in nonhuman primates. Clozapine and olanzapine are atypical antipsychotics designed for clinical use, and deschloroclozapine is a metabolite of clozapine; each has high affinity for DREADD receptors [ 8 , 13 , 18 ].…”

“…All of these actuators have been tested in rodent systems [ 8 , 13 , 16 ] and low-dose clozapine [ 17 ] and deschloroclozapine [ 18 ] have been tested in nonhuman primates. Clozapine and olanzapine are atypical antipsychotics designed for clinical use, and deschloroclozapine is a metabolite of clozapine; each has high affinity for DREADD receptors [ 8 , 13 , 18 ]. Importantly, these drugs can activate DREADD receptors at doses lower than those associated with their therapeutic effects [ 19 – 22 ].…”

Imaging α-synuclein pathologies in animal models and patients with Parkinson’s and related diseases

“…A recombinant adeno associated virus (AAV) was prepared in HEK293T cells by polyethylenimine mediated co-transfection of AAV transfer vector encoding mCherry with rat Synapsin promoter and AAV serotype DJ packaging plasmids, pHelper and pRC-DJ (Cell Biolabs Inc.), as described previously (Nagai et al, 2020). 48 hours after transfection, cells were harvested and lysed in 20 mM HEPES-NaOH, pH 8.0, 150mM NaCl buffer supplemented with 0.5% sodium deoxyholate and 50 units/mL benzonase nuclease (Sigma).…”