| Size | Price | |
|---|---|---|
| Other Sizes |
Purity: ≥98%
| Targets |
TS/thymidine synthase; The primary therapeutic target of doxifluridine is thymidylate synthase (TYMS). As a prodrug, doxifluridine itself is inactive. It requires enzymatic conversion by pyrimidine nucleoside phosphorylase (thymidine phosphorylase) to release the active metabolite 5-fluorouracil (5-FU). The 5-FU is subsequently metabolized to fluorodeoxyuridine monophosphate (FdUMP), which forms a stable ternary complex with thymidylate synthase and methylenetetrahydrofolate, thereby inhibiting DNA synthesis and cell division. Additionally, 5-FU can be incorporated into RNA, interfering with RNA transcription and function.
Doxifluridine is a pyrimidine analogue used as an antineoplastic agent. It is converted to 5-fluorouracil by pyrimidine nucleoside phosphorylase, functioning as an antimetabolite and prodrug. The compound has been shown to have potent antitumor activity against various types of cancer, including colorectal, gastric, and pancreatic cancer. |
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| ln Vitro |
By considerably reducing VEGF expression, doxifluridine (1–10 μM) prevents angiogenesis in FU-MMT-1 cells [1]. At low doses (1 μM), doxifluridine (1-100 μM) slightly increases TSP-1 expression in FU-MMT-1 cells, while at high doses (100 μM), it inhibits TSP-1 expression [1]. In HUVEC cells, dosifluridine (100 μM) can prevent cell division [1].
Beyond its established anticancer activity, recent studies have revealed doxifluridine's potent antibacterial activity. Against clinical multidrug-resistant Staphylococcus aureus (MRSA) isolates, doxifluridine demonstrated minimum inhibitory concentration (MIC) values ranging from 0.5 to 2 µg/mL and minimum bactericidal concentration (MBC) values from 1 to 4 µg/mL. It exhibited rapid bactericidal effects within 8 hours and significantly reduced both biofilm mass and viability of biofilm-embedded bacteria. Furthermore, chequerboard assays showed synergistic interactions with other antibiotics, reducing MIC values by two- to eightfold. Doxifluridine is a fluoropyrimidine derivative and oral prodrug of 5-fluorouracil. In vitro, it is converted to 5-FU by pyrimidine nucleoside phosphorylase, which inhibits thymidylate synthase and disrupts DNA synthesis. The compound displays cytostatic activity against cancer cell lines. |
| ln Vivo |
In BALB cA Jcl-nu mice, doxifluridine (61.55 mg/kg; gavage; single dose) exhibits anti-cancer action. When combined with TNP-470, this anti-cancer activity can be markedly amplified [1]. Ductal acid synthase activity in the DMH-induced ductal carcinoma center can be inhibited at a dose of 200 mg/kg administered intraperitoneally as a single injection [2].
In vivo studies using a Galleria mellonella (wax moth) larvae model infected with MRSA demonstrated that doxifluridine significantly increased the survival rate of infected larvae. In the context of anticancer therapy, studies utilizing mesenchymal stem cells engineered to deliver thymidine phosphorylase to cancer cells showed that doxifluridine treatment (200 µM) induced significant cancer cell death in co-culture systems, with complete eradication of A549 adenocarcinoma cells observed after 5 days at a 2:1 ratio of enzyme-carrying stem cells to cancer cells. Doxifluridine is an antineoplastic agent used in the treatment of various types of cancer. It has been widely investigated in cancer studies, including mouse cachexia, human breast cancer, and mouse bladder cancer. The compound is an oral prodrug that is converted to 5-FU in vivo, providing targeted delivery to tumor tissues. |
| Enzyme Assay |
In vitro tube formation assay.[1]
Tube formation was determined in triplicate in 24-well dishes using an angiogenesis kit according to the manufacturer’s instructions, with a minor modification. FU-MMT-1 cells (1 × 105 cells/well) were co-cultured with human umbilical vein endothelial cells (HUVEC) and fibroblasts, as an in vitro model of tumor angiogenesis, in the presence or absence of the indicated drugs. Doxifluridine, tegafur, 5-FU or TNP-470 was used in this assay.[1] Enzyme immunoassay for VEGF.[1] The concentrations of VEGF in the culture media were determined as previously described using a specific enzyme immunoassay. The medium was obtained on day 9 for each culture. All experiments were repeated independently three times with at least two samples for each treatment. Standard prodrug conversion assays can be conducted using purified thymidine phosphorylase (TYMP) enzyme. A representative protocol: Purified TYMP-coreSA fusion protein is incubated with doxifluridine in an appropriate reaction buffer. The conversion of doxifluridine to 5-FU is monitored over time using high-performance liquid chromatography (HPLC) or UV spectrophotometry. Bradford assays can be used to quantify protein concentration, and enzyme activity is determined by measuring the rate of 5-FU production. In vitro enzyme assays for Doxifluridine typically measure its conversion to 5-fluorouracil by pyrimidine nucleoside phosphorylase. The enzyme is incubated with the compound, and the formation of 5-FU is measured by HPLC or LC-MS. The compound's ability to inhibit thymidylate synthase is assessed using a radiometric assay. IC50 values for cytotoxicity are determined in cancer cell lines. |
| Cell Assay |
Human TSP-1 gene expression in FU-MMT-1 cells and HUVEC.[1]
Low-dose doxifluridine (1 μM) slightly increased TSP-1 expression (P = 0.272) in HUVEC, whereas high-dose doxifluridine (100 μM) reduced TSP-1 expression. Low-dose doxifluridine (1 and 10 μM) significantly increased TSP-1 expression in FU-MMT-1 cells compared with the control group (1 μM, P = 0.041; 10 μM, P = 0.0089).[1] Cytotoxicity assay in HUVEC.[1] Doxifluridine (100 μM) reduced the proliferation of HUVEC compared with the control group, although this did not reach statistical significance (P = 0.079). Meanwhile, TNP-470 (10 and 100 μM) significantly inhibited the proliferation of HUVEC compared with the control group (10 μM, P = 0.041; 100 μM, P = 0.011). However, lower concentrations of doxifluridine (10 μM) were not cytotoxic against HUVEC. Cytotoxicity of doxifluridine is evaluated using cancer cell lines that express varying levels of thymidine phosphorylase. A representative protocol: Cells (e.g., A549 adenocarcinoma cells) are seeded in 96-well plates. Doxifluridine is dissolved in culture medium and added at concentrations ranging from 0 to 200 µM. After 72-120 hours of incubation, cell viability is assessed using MTT or CCK-8 assays. For co-culture studies, cells are mixed with TYMP-expressing cells or enzyme-conjugated carriers prior to doxifluridine addition. Antibacterial activity is assessed via broth microdilution to determine MIC and MBC values. Cell-based cytotoxicity assays for Doxifluridine involve treating various cancer cell lines (e.g., colorectal, gastric, pancreatic) with increasing concentrations of the compound. Cell viability is assessed after 48-72 hours using MTT, CellTiter-Glo, or clonogenic assays. IC50 values are calculated from dose-response curves. The compound's ability to inhibit cell proliferation and induce apoptosis is evaluated. Thymidylate synthase activity is measured in treated cells. |
| Animal Protocol |
In vivo treatments.[1]
The mice were injected subcutaneously with 2 × 105 FU-MMT-1 cells in 0.2 mL of vehicle. Mice that developed tumors measuring 5–10 mm in diameter by day 7 were randomly separated into five groups (n = 5–6/group) and were treated as follows: (i) doxifluridine alone at the MTD of 184.65 mg/kg (the MTD was calculated by referring to the initial in vivo study of doxifluridine, which was administered by gavage); (ii) metronomic doxifluridine at a dose of 61.55 mg/kg; (iii) TNP-470 alone; (iv) metronomic doxifluridine in combination with TNP-470; and (v) control (untreated). Treatments were continued for 8 weeks. Doxifluridine was administered by gavage once daily for 5 consecutive days/week with no prolonged breaks. TNP-470 was injected subcutaneously at a dose of 30 mg/kg three times per week. Tumor growth was monitored by measuring the size of the tumor twice/week, and was calculated as V = length × width2/2. Bodyweight of the mice was measured weekly. For evaluating anticancer efficacy, murine xenograft models are commonly used. A protocol using a Galleria mellonella larvae model has been described for assessing antibacterial activity: Larvae are injected with MRSA inoculum, followed by doxifluridine treatment. Survival rates are monitored over 3-5 days, and bacterial burdens are quantified by colony-forming unit (CFU) counts from larval homogenates. In vivo animal studies for Doxifluridine typically involve administering the compound to mice bearing tumor xenografts. The compound is given orally at various doses. Tumor growth inhibition is monitored over several weeks. The compound's conversion to 5-FU in tumor tissue is measured. Pharmacodynamic endpoints include tumor volume, weight, and histology. Survival is also assessed. |
| ADME/Pharmacokinetics |
Doxifluridine is well-absorbed after oral administration. In a pharmacokinetic study involving 20 colorectal cancer patients, following oral administration of doxifluridine (1200 mg/m² daily for 5 days), the area under the curve (AUC) of doxifluridine ranged between 72.2 and 74.5 mmol·h/L, with a Cmax remaining in a narrow band of 67.1 to 68.3 mmol/L from day 1 to day 5. In contrast, the AUC of its active metabolite 5-FU increased from 5.46 to 7.52 mmol·h/L over the same period. A significant correlation between the AUC of doxifluridine and 5-FU was observed (P < 0.001), and 5-FU levels remained detectable for a longer duration compared to classical 5-FU therapy. The drug is primarily metabolized in the liver and excreted via the kidneys.
Pharmacokinetic properties of Doxifluridine are characterized by its oral bioavailability. The compound is absorbed following oral administration and converted to 5-fluorouracil by pyrimidine nucleoside phosphorylase. The conversion occurs preferentially in tumor tissues due to higher enzyme activity. The half-life of the compound is relatively short. 5-FU is metabolized in the liver and excreted through renal pathways. |
| Toxicity/Toxicokinetics |
In clinical use, doxifluridine is generally well-tolerated but can cause adverse effects. The most common toxicities include gastrointestinal reactions (diarrhea, nausea, vomiting, anorexia), hematologic effects (leukopenia, hemoglobin reduction, thrombocytopenia), and hepatotoxicity (elevated liver enzymes GOT, GPT, ALP, bilirubin). Other reported adverse reactions include stomatitis, pigmentation, alopecia, fever, and occasional CNS effects (dizziness, headache, somnolence). Serious but less common toxicities include severe enteritis (hemorrhagic, ischemic, or necrotizing enteritis) requiring prompt discontinuation, and myelosuppression requiring regular blood monitoring. Doxifluridine is contraindicated in pregnant/nursing women and patients receiving sorivudine therapy. Safety evaluations confirmed that doxifluridine did not exhibit hemolytic toxicity or significant cytotoxicity in vitro.
Toxicological profile of Doxifluridine is related to its conversion to 5-fluorouracil. Common side effects include myelosuppression, gastrointestinal toxicity (nausea, vomiting, diarrhea), mucositis, and hand-foot syndrome. The compound's toxicity is dose-dependent. Close monitoring of blood cell counts and liver function is required during treatment. The safety profile is consistent with fluoropyrimidine antineoplastic agents. |
| References |
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| Additional Infomation |
Doxifluridine is a 5'-deoxynucleoside of pyrimidine, a derivative of 5-fluorouracil, with its 5'-hydroxyl group replaced by a hydrogen atom. It is an oral prodrug of the antitumor drug 5-fluorouracil. Doxifluridine is designed to avoid the rapid degradation of 5-fluorouracil by dihydropyrimidine dehydrogenases in the intestinal wall; it is converted to 5-fluorouracil by pyrimidine nucleoside phosphorylases. It has multiple effects including antimetabolism, antitumor activity, and prodrug activity. It is an organofluorine compound and also a 5'-deoxynucleoside of pyrimidine. Doxifluridine has been studied for the treatment of gastric cancer. Doxifluridine is a fluoropyrimidine derivative and an oral prodrug of the antitumor drug 5-fluorouracil (5-FU). Doxifluridine is designed to avoid the rapid degradation of 5-fluorouracil by dihydropyrimidine dehydrogenases in the intestinal wall; it is converted to 5-fluorouracil by pyrimidine nucleoside phosphorylases. 5-Fluorouracil interferes with DNA synthesis and subsequent cell division by reducing normal thymidine production and interferes with RNA transcription by competing with uridine triphosphate for incorporation sites in the RNA chain.
Doxifluridine is an oral prodrug of 5-fluorouracil used as an antineoplastic agent. It has been investigated for the treatment of various cancers, including colorectal, gastric, and pancreatic cancer. The compound is a pyrimidine analogue. It is not widely used in clinical practice due to the availability of other fluoropyrimidine formulations. |
| Molecular Formula |
C9H11FN2O5
|
|---|---|
| Molecular Weight |
246.19
|
| Exact Mass |
246.065
|
| Elemental Analysis |
C, 43.91; H, 4.50; F, 7.72; N, 11.38; O, 32.49
|
| CAS # |
3094-09-5
|
| Related CAS # |
Doxifluridine-d2;84258-25-3
|
| PubChem CID |
18343
|
| Appearance |
White to off-white solid
|
| Density |
1.6±0.1 g/cm3
|
| Melting Point |
188-192 °C(lit.)
|
| Index of Refraction |
1.605
|
| LogP |
-0.96
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
17
|
| Complexity |
399
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
C[C@@H]1[C@H]([C@H]([C@H](N2C(NC(C(F)=C2)=O)=O)O1)O)O
|
| InChi Key |
ZWAOHEXOSAUJHY-ZIYNGMLESA-N
|
| InChi Code |
InChI=1S/C9H11FN2O5/c1-3-5(13)6(14)8(17-3)12-2-4(10)7(15)11-9(12)16/h2-3,5-6,8,13-14H,1H3,(H,11,15,16)/t3-,5-,6-,8-/m1/s1
|
| Chemical Name |
1-((2R,3R,4S,5R)-3,4-dihydroxy-5-methyltetrahydrofuran-2-yl)-5-fluoropyrimidine-2,4(1H,3H)-dione
|
| Synonyms |
Flutron; Furtulon; 5'-Deoxy-5-fluorouridine; 5-fluoro-5'-deoxyuridine; Furtulon; 3094-09-5; doxifluridine
|
| 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 (In Vitro) |
DMSO : ≥ 100 mg/mL (~406.19 mM)
DMF : 100 mg/mL (~406.19 mM) H2O : ~20 mg/mL (~81.24 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 27.5 mg/mL (111.70 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.0619 mL | 20.3095 mL | 40.6190 mL | |
| 5 mM | 0.8124 mL | 4.0619 mL | 8.1238 mL | |
| 10 mM | 0.4062 mL | 2.0310 mL | 4.0619 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.
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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