Clozapine N-oxide (CNO)
(Synonyms: 氯氮平N-氧化物) 目录号 : GC10822
Clozapine N-oxide（CNO）是一种强效的多巴胺拮抗剂，也是一种选择性肌动蛋白M4受体（EC50=11 nM）激动剂。
Cas No.:34233-69-7
Sample solution is provided at 25 µL, 10mM.
Clozapine N-oxide(CNO) is a potent dopamine antagonist and also a potent and selective muscarinic M4 receptor (EC50=11 nM) agonist[5，6].Clozapine N-oxide also is a major metabolite of Clozapine and a human muscarinic designer receptors (DREADDs) agonist[1,2]. Clozapine N-oxide activates the DREADD receptor hM3Dq and hM4Di. Clozapine N-oxide(CNO) is a pharmacologically inert molecule that lacks a significant (<1 μm) affinity for the receptor[3]. Clozapine N-oxide(CNO) is highly bioavailable in rodents and humans[4]
Clozapine N-oxide(CNO) significantly reduces the density of the 5-HT2 receptor in rat cortical cells, A significant decrease was found in primary cortical cells for 5-HT(2) receptor density and 5-HT(2A) receptor mRNA levels[7].
In both β-R-q and β-R-s Tg mice, Clozapine N-oxide(CNO) treatment resulted in a dose-dependent decrease in blood glucose levels and increased plasma insulin concentrations, demonstrating that the degree of β cell G protein signaling could be titrated according to the Clozapine N-oxide(CNO) dose administered[8].Oral Clozapine N-oxide(CNO) administration via drinking water to mice expressing rM3Dq in pancreatic β-cells led to robust metabolic phenotypes, indicating that significant amounts of Clozapine N-oxide(CNO) are absorbed from the gastrointestinal tract[9].It created transgenic mice expressing an evolved G protein-coupled receptor (hM3Dq) selectively activated by the pharmacologically inert, orally bioavailable drug Clozapine N-oxide (CNO).Local field potential and single-neuron recordings revealed that peripheral administration of Clozapine N-oxide(CNO) activated hippocampal neurons selectively in hM3Dq-expressing mice. Behavioral correlates of neuronal activation included increased locomotion, stereotypy, and limbic seizures[10].
References:
[1]: Wess J, Nakajima K, et,al. Novel designer receptors to probe GPCR signaling and physiology. Trends Pharmacol Sci. 2013 Jul;34(7):385-92. doi: 10.1016/j.tips.2013.04.006. Epub 2013 Jun 13. PMID: 23769625; PMCID: PMC3758874.
[2]: Manvich DF, Webster KA, et,al. The DREADD agonist clozapine N-oxide (CNO) is reverse-metabolized to clozapine and produces clozapine-like interoceptive stimulus effects in rats and mice. Sci Rep. 2018 Mar 1;8(1):3840. doi: 10.1038/s41598-018-22116-z. PMID: 29497149; PMCID: PMC5832819.
[3]: Weiner DM, Meltzer HY, et,al. The role of M1 muscarinic receptor agonism of N-desmethylclozapine in the unique clinical effects of clozapine. Psychopharmacology (Berl). 2004 Dec;177(1-2):207-16. doi: 10.1007/s00213-004-1940-5. Epub 2004 Jul 16. PMID: 15258717.
[4]: Bender D, Holschbach M, et,al. Synthesis of n.c.a. carbon-11 labelled clozapine and its major metabolite clozapine-N-oxide and comparison of their biodistribution in mice. Nucl Med Biol. 1994 Oct;21(7):921-5. doi: 10.1016/0969-8051(94)90080-9. PMID: 9234345.
[5]: Silva RR, Parreiras-E-Silva LT, et,al. Evaluation of Functional Selectivity of Haloperidol, Clozapine, and LASSBio-579, an Experimental Compound With Antipsychotic-Like Actions in Rodents, at G Protein and Arrestin Signaling Downstream of the Dopamine D2 Receptor. Front Pharmacol. 2019 Jun 4;10:628. doi: 10.3389/fphar.2019.00628. PMID: 31214037; PMCID: PMC6558205.
[6]: Zorn SH, Jones SB, et,al. Clozapine is a potent and selective muscarinic M4 receptor agonist. Eur J Pharmacol. 1994 Nov 15;269(3):R1-2. doi: 10.1016/0922-4106(94)90047-7. PMID: 7895765.
[7]: Heiser P, Schulte E, et,al. Effects of clozapine and its metabolites on the 5-HT2 receptor system in cortical and hippocampal cells in vitro. Prog Neuropsychopharmacol Biol Psychiatry. 2004 Mar;28(2):297-302. doi: 10.1016/j.pnpbp.2003.10.008. PMID: 14751426.
[8]: Guettier JM, Gautam D, et,al. A chemical-genetic approach to study G protein regulation of beta cell function in vivo. Proc Natl Acad Sci U S A. 2009 Nov 10;106(45):19197-202. doi: 10.1073/pnas.0906593106. Epub 2009 Oct 26. PMID: 19858481; PMCID: PMC2767362.
[9]: Jain S, Ruiz de Azua I, et,al. Chronic activation of a designer G(q)-coupled receptor improves β cell function. J Clin Invest. 2013 Apr;123(4):1750-62. doi: 10.1172/JCI66432. Epub 2013 Mar 8. PMID: 23478411; PMCID: PMC3613926.
[10]:Alexander GM, Rogan SC, et,al. Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled receptors. Neuron. 2009 Jul 16;63(1):27-39. doi: 10.1016/j.neuron.2009.06.014. PMID: 19607790; PMCID: PMC2751885.
| Cell experiment [1]: | |
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Cell lines |
Primary rat cortical cells |
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Preparation Method |
Receptor parameters (density affinity and mRNA levels) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA levels in rat primary cortical cells after 24 h of Clozapine N-oxide(CNO) (200/400 ng/ml) treatment under in vitro conditions |
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Reaction Conditions |
Clozapine N-oxide(CNO) (200/400 ng/ml)for 24h |
|
Applications |
A significant decrease was found in primary cortical cells for 5-HT(2) receptor density (Clozapine N-oxide(CNO) 200/ 400/ 200 and Cloz-N-oxide 200 vs. control) and 5-HT(2A) receptor mRNA levels (Clozapine N-oxide(CNO) 200 vs. control). |
| Animal experiment [2]: | |
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Animal models |
β-RQ and β-RS Tg mice |
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Preparation Method |
To determine the in vivo effects of acute activation of β-cell G q /11 or G S signaling on glucose homeostasis, mice were injected with increased doses of Clozapine N-oxide(CNO) (0.0001-10 mg/kg) and blood glucose levels were monitored for 2 h. Before the experiment began, all mice were free to eat and were given. |
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Dosage form |
0.0001-10 mg/kg Clozapine N-oxide(CNO) for 2 h |
|
Applications |
In both β-R-q and β-R-s Tg mice, Clozapine N-oxide(CNO) treatment resulted in a dose-dependent decrease in blood glucose levels and increased plasma insulin concentrations, demonstrating that the degree of β cell G protein signaling could be titrated according to the Clozapine N-oxide(CNO) dose administered. |
|
References: [1]: Heiser P, Schulte E, et,al. Effects of clozapine and its metabolites on the 5-HT2 receptor system in cortical and hippocampal cells in vitro. Prog Neuropsychopharmacol Biol Psychiatry. 2004 Mar;28(2):297-302. doi: 10.1016/j.pnpbp.2003.10.008. PMID: 14751426. |
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| Cas No. | 34233-69-7 | SDF | |
| 别名 | 氯氮平N-氧化物 | ||
| 化学名 | 3-chloro-6-(4-methyl-4-oxidopiperazin-4-ium-1-yl)-5H-benzo[b][1,4]benzodiazepine | ||
| Canonical SMILES | C[N+]1(CCN(CC1)C2=C3C=CC=CC3=NC4=C(N2)C=C(C=C4)Cl)[O-] | ||
| 分子式 | C18H19ClN4O | 分子量 | 342.82 |
| 溶解度 | ≥ 17.15 mg/mL in DMSO, ≥ 49 mg/mL in Water with gentle warming This product is unstable in solution and it is recommended to prepare and use it immediately. |
储存条件 | Store at -20°C |
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.917 mL | 14.5849 mL | 29.1698 mL |
| 5 mM | 0.5834 mL | 2.917 mL | 5.834 mL |
| 10 mM | 0.2917 mL | 1.4585 mL | 2.917 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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Related Biological Data
Specific manipulations with the VTA DA neuron activity alter depression-like behaviors The immobility time of the AAV-DIO-hM4D-mCherryinjected (C) and the AAV-DIO-mCherry-injected (D) mice in the TST (i) or the FST (ii) were measured before and 15 min after the administration of CNO.
One week after the injection of AAV-DIO-hM4D(Gi)-mCherry/AAV-DIO-hM3D-mCherry or AAV-DIO-mCherry and before the behavioral tests, clozapine N-oxide, CNO (3.3 mg kg-1, i.p.)(GLPBIO) was scheduled 15 minutes prior to the test.
Cell reports 37.5 (2021): 109936. PMID: 34731609 IF: 9.424 -
Related Biological Data
Chemogenetic activation of MCH neurons induced anxiety-like behaviors in mice.(C) Effects of 0.3 mg/kg and 0.15 mg/kg CNO on OFT, EPM and MBT in wild type mice. (n = 6–8 mice per group).
On the experiment day, CNO (0.15 mg/kg)(GLPBIO) was injected intraperitoneally to induce the activation of hM3Dqpositive MCH neurons in the LHA 20 min prior to the anxiety behaviors tests.
Frontiers in Pharmacology 13 (2022): 906057. PMID: 36016574 IF: 5.6