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N4-Methylcytidine (N(4)-methylcytidine)

Cat No.:V55279 Purity: ≥98%
N4-Methylcytidine (Nsc518744) is a cytosine nucleoside analog.
N4-Methylcytidine (N(4)-methylcytidine)
N4-Methylcytidine (N(4)-methylcytidine) Chemical Structure CAS No.: 10578-79-7
Product category: Nucleoside Antimetabolite/Analog
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
50mg
100mg
250mg
Other Sizes
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Product Description
N4-Methylcytidine (Nsc518744) is a cytosine nucleoside analog. Cytidine analogs have a mechanism of inhibiting DNA methyltransferase and have potential anti-metabolic and anti-tumor activities.
N4-Methylcytidine (m4C) is a cytidine nucleoside analog and a significant RNA modification that plays crucial roles in various biological processes, including gene expression regulation, RNA stability, and the fidelity of reverse transcription. Cytidine analogs have a mechanism of inhibiting DNA methyltransferases (such as Zebularine) and have potential anti-metabolic and anti-tumor activities. It is a research-grade compound not approved for human therapeutic use.
Biological Activity I Assay Protocols (From Reference)
Targets
DNA methyltransferases (DNMTs); RNA metabolism pathways. N4-Methylcytidine is a cytidine nucleoside analog that inhibits DNA methyltransferases. Cytidine analogs have a mechanism of inhibiting DNA methyltransferase and have potential anti-metabolic and anti-tumor activities. The compound also plays roles in RNA modification, affecting gene expression regulation and RNA stability.
ln Vitro
N4-Methylcytidine demonstrates activity as a cytidine nucleoside analog in cellular assays. It inhibits DNA methyltransferases and has potential anti-metabolic and anti-tumor activities. It is used as a research tool in epigenetic studies. Detailed IC50 values have not been disclosed.
ln Vivo
In vivo activity of N4-Methylcytidine has not been extensively characterized. As a cytidine analog, it would be expected to have anti-tumor activity in animal models. The compound is primarily used in research on epigenetic regulation, DNA methylation dynamics, and tumor-associated metabolic reprogramming.
Enzyme Assay
In vitro enzyme assays for DNA methyltransferase inhibition are performed using recombinant DNMT enzymes and DNA substrates in the presence of S-adenosylmethionine (SAM). N4-Methylcytidine is incubated at varying concentrations with the enzyme and substrate. Methylation is detected by radioactivity or immunoassay. IC50 values are calculated from dose-response curves.
Cell Assay
Cells (e.g., cancer cell lines) are cultured and treated with N4-Methylcytidine at concentrations typically ranging from 1–100 μM for 24–72 hours. DNA methylation levels are assessed by methylation-specific PCR or bisulfite sequencing. DNMT inhibition is assessed by measuring global DNA methylation or specific gene methylation. Cell viability and proliferation are evaluated by standard assays.
Animal Protocol
In vivo animal studies for N4-Methylcytidine have not been reported in publicly available literature. The compound is primarily used as an in vitro research tool for epigenetic studies. Future studies in animal models of cancer would be needed to evaluate in vivo efficacy.
ADME/Pharmacokinetics
Pharmacokinetic data for N4-Methylcytidine have not been published. The compound has a molecular weight of 257.24 and formula C10H15N3O5. As a cytidine analog, it is expected to be taken up by cells via nucleoside transporters. In vivo PK parameters such as half-life, bioavailability, and tissue distribution have not been characterized.
Toxicity/Toxicokinetics
Toxicology data for N4-Methylcytidine have not been published. The compound is for research use only and not approved for human therapeutic applications. No systematic toxicological evaluation has been performed. Standard safety precautions for handling nucleoside analogs should be observed.
References

[1]. Mechanism of inhibition of DNA methyltransferases by cytidine analogs in cancer therapy. Cancer Biol Ther. 2004 Nov;3(11):1062-8.

Additional Infomation
N4-methylcytidine is a pyrimidine nucleoside.
N4-Methylcytidine (CAS: 10578-79-7, molecular formula C10H15N3O5, molecular weight 257.24) is a cytidine nucleoside analog and RNA modification. It inhibits DNA methyltransferases and has potential anti-metabolic and anti-tumor activities. It is used in research on epigenetic regulation and DNA methylation dynamics. It is not in clinical trials and has no regulatory approval.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C10H15N3O5
Molecular Weight
257.24
Exact Mass
257.101
CAS #
10578-79-7
PubChem CID
101436
Appearance
White to off-white solid powder
Density
1.69g/cm3
Boiling Point
504.2ºC at 760mmHg
Flash Point
258.7ºC
Vapour Pressure
2.82E-12mmHg at 25°C
Index of Refraction
1.694
LogP
-1.7
Hydrogen Bond Donor Count
4
Hydrogen Bond Acceptor Count
5
Rotatable Bond Count
3
Heavy Atom Count
18
Complexity
397
Defined Atom Stereocenter Count
4
SMILES
CNC1=NC(=O)N(C=C1)[C@H]2[C@@H]([C@@H]([C@H](O2)CO)O)O
InChi Key
LZCNWAXLJWBRJE-ZOQUXTDFSA-N
InChi Code
InChI=1S/C10H15N3O5/c1-11-6-2-3-13(10(17)12-6)9-8(16)7(15)5(4-14)18-9/h2-3,5,7-9,14-16H,4H2,1H3,(H,11,12,17)/t5-,7-,8-,9-/m1/s1
Chemical Name
1-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-4-(methylamino)pyrimidin-2-one
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

Note: This product requires protection from light (avoid light exposure) during transportation and storage.
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)
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
Solubility (In Vivo)
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

Injection Formulations
(e.g. IP/IV/IM/SC)
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution 50 μL Tween 80 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO 400 μLPEG300 50 μL Tween 80 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).
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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO 100 μLPEG300 200 μL castor oil 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10: 10 : 80 (i.e. 100 μL Ethanol 100 μL Cremophor 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH 400 μLPEG300 50 μL Tween 80 450 μL Saline)


Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).
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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders


Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 3.8874 mL 19.4371 mL 38.8742 mL
5 mM 0.7775 mL 3.8874 mL 7.7748 mL
10 mM 0.3887 mL 1.9437 mL 3.8874 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:

  • Calculate the Mass of a compound required to prepare a solution of known volume and concentration
  • Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
  • Calculate the Concentration of a solution resulting from a known mass of compound in a specific volume
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)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • 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)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • 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.

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