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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
Official Supplier of:
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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)
DMSO: ~50 mg/mL (~194.4 mM; with ultrasonication)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.72 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and 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 (9.72 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 DMSO stock solution (25.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well.
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 3: ≥ 2.5 mg/mL (9.72 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 DMSO stock solution (25.0 mg/mL) to 900 μL of corn oil and mix well.

 (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.

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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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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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