Clozapine N-oxide

Alias: Clozapine N-oxide
Cat No.:V9293 Purity: ≥98%
Clozapine N-oxide is a major metabolite of Clozapine and an agonist of human designer muscarinic receptors (DREADDs).
Clozapine N-oxide Chemical Structure CAS No.: 34233-69-7
Product category: Dopamine Receptor
This product is for research use only, not for human use. We do not sell to patients.
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Other Forms of Clozapine N-oxide:

  • Clozapine (HF 1854)
  • Clozapine N-oxide diHCl
Official Supplier of:
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Purity & Quality Control Documentation

Purity: ≥98%

Product Description
Clozapine N-oxide is a major metabolite of Clozapine and an agonist of human designer muscarinic receptors (DREADDs). Clozapine N-oxide activates the DREADD receptors hM3Dq and hM4Di. Clozapine N-oxide can cross the BBB (blood-brain barrier). Clozapine is a potent dopamine antagonist and a potent and specific muscarinic M4 receptor (EC50 of 11 nM) agonist.
Biological Activity I Assay Protocols (From Reference)
Targets
DREADD; Dopamine receptor; muscarinic M4 receptor
ln Vitro
CNO is a synthetic small molecule actuator for DREADDs that muscarinic acetylcholinergic.
ln Vivo
Clozapine N-oxide (CNO), an agonist of the human muscarinic designer receptor (DREADDs), is transformed into clozapine and exhibits behavioral effects resembling those of clozapine in both mice and rats. This highlights the necessity of using suitable control groups in research involving DREADDs.[4]
Animal Protocol
standard laboratory rats and mice
1.0, 3.2, 10.0 mg/kg for rats; 1.25, 2.5, 5.0, 10.0, 20.0 mg/kg for mice
i.p.
References

[1]. Sci Rep . 2018 Mar 1;8(1):3840.

[2]. Front Pharmacol . 2019 Jun 4:10:628.

[3]. Eur J Pharmacol . 1994 Nov 15;269(3):R1-2.

[4]. Sci Rep . 2018 Mar 1;8(1):3840.

[5]. MethodsX . 2018 Mar 23:5:257-267.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C18H19CLN4O
Molecular Weight
342.82266
Exact Mass
342.12
Elemental Analysis
C, 63.06; H, 5.59; Cl, 10.34; N, 16.34; O, 4.67
CAS #
34233-69-7
Related CAS #
Clozapine; 5786-21-0; Clozapine N-oxide dihydrochloride; 2250025-93-3; 54241-01-9 (HCl); 34233-69-7 (N-oxide); 5786-21-0
Appearance
Solid powder
SMILES
C[N+]1(CCN(CC1)C2=NC3=C(C=CC(=C3)Cl)NC4=CC=CC=C42)[O-]
InChi Key
OGUCZBIQSYYWEF-UHFFFAOYSA-N
InChi Code
InChI=1S/C18H19ClN4O/c1-23(24)10-8-22(9-11-23)18-14-4-2-3-5-15(14)20-16-7-6-13(19)12-17(16)21-18/h2-7,12,20H,8-11H2,1H3
Chemical Name
3-chloro-6-(4-methyl-4-oxidopiperazin-4-ium-1-yl)-11H-benzo[b][1,4]benzodiazepine
Synonyms
Clozapine N-oxide
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO: 68~100 mg/mL (198.4~291.7 mM)
Water: ~68 mg/mL
Ethanol: ~8 mg/mL
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.29 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: ≥ 2.5 mg/mL (7.29 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (7.29 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.


Solubility in Formulation 4: ≥ 0.5 mg/mL (1.46 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 5: ≥ 0.5 mg/mL (1.46 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

Solubility in Formulation 6: 5%DMSO+ 40%PEG300+ 5%Tween 80+ 50%ddH2O: 3.4mg/ml (9.92mM)

Solubility in Formulation 7: 2.44 mg/mL (7.12 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.9170 mL 14.5849 mL 29.1698 mL
5 mM 0.5834 mL 2.9170 mL 5.8340 mL
10 mM 0.2917 mL 1.4585 mL 2.9170 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
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  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
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  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
  • To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.
Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

  • Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
  • Click the “Calculate” button
  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • Oxidation of clozapine to clozapine N-oxide (CNO) using Oxone. MethodsX . 2018 Mar 23:5:257-267.
  • ORTEP representation of the molecular structure of CNO. MethodsX . 2018 Mar 23:5:257-267.
  • CNO produces clozapine-like interoceptive stimulus effects in standard laboratory rats and mice. Sci Rep . 2018 Mar 1;8(1):3840.
  • CNO is converted to clozapine in standard laboratory rats and mice. Sci Rep . 2018 Mar 1;8(1):3840.
  • CNO produces clozapine-like interoceptive stimulus effects in standard laboratory rats and mice. Sci Rep . 2018 Mar 1;8(1):3840.
  • CNO is converted to clozapine in standard laboratory rats and mice. Sci Rep . 2018 Mar 1;8(1):3840.
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