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| 1mg |
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| 5mg |
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| Targets |
Thymidine-15N2 targets metabolic enzymes and pathways involved in DNA synthesis. As a DNA synthesis inhibitor, it acts on the cellular replication machinery by blocking the G1/S transition. The compound is also an endogenous metabolite that naturally participates in nucleic acid metabolism. Through its role as a thymidine analog, it interferes with the normal progression of DNA replication, making it a valuable tool for studying cell cycle regulation and the mechanisms of DNA synthesis inhibition.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantification throughout the drug development process. Due to its potential to alter the pharmacokinetic and metabolic characteristics of medications, deuteration has drawn attention[1].
In vitro studies demonstrate that Thymidine functions as a DNA synthesis inhibitor that blocks cells at the G1/S boundary prior to DNA replication. The compound is used as a cell synchronizing agent in tissue culture applications. When applied to cultured cells, it arrests the cell cycle at the G1/S checkpoint, allowing researchers to study cell cycle progression and DNA replication mechanisms. This property has made thymidine a standard tool in cell biology for synchronizing cell populations in vitro. |
| ln Vivo |
In vivo, isotopically labeled thymidine compounds are primarily used as tracers for metabolic studies. The stable isotope labeling allows for the tracking of the compound within biological systems. Due to its role as a DNA precursor, labeled thymidine can be incorporated into newly synthesized DNA, enabling studies of cell proliferation and turnover in living organisms. This makes it valuable for investigating tissue regeneration, tumor growth dynamics, and the pharmacokinetics of nucleoside analogs in animal models.
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| Enzyme Assay |
The binding properties of Thymidine-15N2 to its molecular targets can be studied using surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) to measure interactions with DNA polymerases and other nucleotide-binding proteins. Competitive binding assays using radiolabeled or fluorescently labeled thymidine can determine binding affinities. Typical assay conditions involve physiological buffer systems (PBS or Tris-HCl, pH 7.4) at 25degC or 37degC with varying concentrations of the labeled compound to calculate dissociation constants (Kd).
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| Cell Assay |
Standard cell synchronization protocols using thymidine involve the double thymidine block method. Cells are cultured in appropriate growth medium and treated with 2 mM thymidine for 18-24 hours to arrest cells at the G1/S boundary. After removal of thymidine by washing with fresh medium, cells are allowed to progress through the cell cycle for 9-12 hours before a second thymidine block is applied. Cell cycle distribution is monitored by flow cytometry following propidium iodide staining to confirm synchronization efficiency.
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| Animal Protocol |
For in vivo pharmacokinetic and biodistribution studies, Thymidine-15N2 can be administered to rodents via intravenous, intraperitoneal, or oral routes at doses typically ranging from 10-100 mg/kg. Blood samples are collected at various time points post-administration, and tissues of interest (liver, kidney, spleen, etc.) are harvested for analysis. The labeled compound is extracted from biological matrices and quantified using LC-MS/MS, taking advantage of the distinct mass shift conferred by the 15N isotopes to distinguish the labeled compound from endogenous thymidine.
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| ADME/Pharmacokinetics |
As a stable isotope-labeled compound, Thymidine-15N2 serves primarily as an analytical tracer. Its pharmacokinetic properties largely mirror those of unlabeled thymidine. Thymidine is rapidly cleared from circulation with a short half-life in rodents (typically 10-30 minutes). It undergoes extensive metabolism via thymidine phosphorylase and other nucleoside-metabolizing enzymes, producing thymine and deoxyribose-1-phosphate. The compound distributes widely throughout the body, with particular uptake in proliferating tissues such as bone marrow and intestinal epithelium.
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| Toxicity/Toxicokinetics |
Thymidine itself is generally well-tolerated in research settings. At the concentrations used for cell synchronization (2 mM), no significant cytotoxicity is observed in most cell types during short-term exposure. In animal studies, high doses may cause reversible bone marrow suppression due to its effects on DNA synthesis. The isotopically labeled version is not expected to exhibit altered toxicity profiles compared to the unlabeled compound. Standard laboratory safety precautions should be followed when handling the compound.
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| References | |
| Additional Infomation |
Thymidine-15N2 is not intended for clinical use and has not been approved as a therapeutic drug; it is exclusively a research tool for stable isotope tracing studies. The compound can be detected by various analytical techniques such as mass spectrometry, where the two incorporated 15N atoms create a distinct mass shift. This version of thymidine incorporates two nitrogen-15 atoms (positions 1 and 3 of the pyrimidine ring), making it essential for precise quantification in metabolic flux analysis. Due to its potential to alter pharmacokinetic and metabolic characteristics, isotopic labeling has drawn significant attention in drug development research.
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| Molecular Formula |
C10H1415N2O5
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| Related CAS # |
Thymidine;50-89-5
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| Appearance |
Typically exists as solid at room temperature
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| 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
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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 : PEG300 ：Tween 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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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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