Pharmacokinetic and pharmacodynamic actions of clozapine-N-oxide, clozapine, and compound 21 in DREADD-based chemogenetics in mice
Pharmacokinetic and pharmacodynamic actions of clozapine-N-oxide, clozapine, and compound 21 in DREADD-based chemogenetics in mice
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DOI：
10.1038/s41598-019-41088-2
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发表时间：
2019-03-14
影响因子：
4.6
通讯作者：
Pekcec, Anton
中科院分区：
文献类型：
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作者：
Jendryka, Martin;Palchaudhuri, Monika;Pekcec, Anton
由氯氮平N-氧化物（CNO）门控的由设计药物专门激活的毒蕈碱设计受体（DREADD）允许在体内遗传特异性细胞类型中选择性地激活G蛋白级联。在此，我们比较了CNO、氯氮平（CLZ）和化合物21（Cmpd-21）在抑制性DREADD人Gi偶联M4毒蕈碱受体（hM 4Di）处的药代动力学、脱靶效应和功效。CLZ的半数最大有效浓度（EC 50）显著低于CNO（8.1 nM）（0.42 nM）; Cmpd-21居中（2.95 nM）。CNO在小鼠体内被转化为CLZ，CLZ在脑组织中积累。然而，CNO本身也进入大脑，并且游离脑脊液（CSF）水平在直接激活hM 4Di的范围内，而游离（CSF）CLZ水平保持低于检测限。此外，直接注射的CLZ强烈转化为其非活性代谢物，去甲氯氮平。CMPD-21显示出上级的脑渗透性和持久存在性。虽然我们确定了广泛的CNO和Cmpd-21脱靶，几乎没有任何非特异性的行为影响之间的参数评估的5-choiceserial-reaction-time任务。我们的结果表明，CNO（3-5 mg/kg）和Cmpd-21（0.4-1 mg/kg）是合适的DREADD激动剂，在腹膜内应用后最迟15分钟有效，但两者都需要受试者之间的非特异性作用对照。
Muscarinic Designer Receptors Exclusively Activated by Designer Drugs (DREADD) gated by clozapineN-oxide (CNO) allow selective G-protein cascade activation in genetically specified cell-types in vivo. Here we compare the pharmacokinetics, off-target effects and efficacy of CNO, clozapine (CLZ) and compound 21 (Cmpd-21) at the inhibitory DREADD human Gi-coupled M4 muscarinic receptor (hM4Di). The half maximal effective concentration (EC50) of CLZ was substantially lower (0.42 nM) than CNO (8.1 nM); Cmpd-21 was intermediate (2.95 nM). CNO was back-converted to CLZ in mice, and CLZ accumulated in brain tissue. However, CNO itself also entered the brain, and free cerebrospinal fluid (CSF) levels were within the range to activate hM4Di directly, while free (CSF) CLZ levels remained below the detection limit. Furthermore, directly injected CLZ was strongly converted to its pharmacologically active metabolite, norclozapine. Cmpd-21 showed a superior brain penetration and long-lasting presence. Although we identified a wide range of CNO and Cmpd-21 off-targets, there was hardly any nonspecific behavioural effects among the parameters assessed by the 5-choiceserial-reaction-time task. Our results suggest that CNO (3-5 mg/kg) and Cmpd-21 (0.4-1 mg/kg) are suitable DREADD agonists, effective at latest 15 min after intraperitoneal application, but both require between-subject controls for unspecific effects.