5-Fluorouridine
(Synonyms: 5-氟尿嘧啶核苷) 目录号 : GC38141
5-Fluorouridine是5-氟尿嘧啶（5-Fluorouracil）的核糖核苷酸代谢物，具有抗癌活性。5-Fluorouridine可在体内和体外标记细胞的活性转录位点，用于新生RNA的免疫检测。
Cas No.:316-46-1
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导，请根据您的具体需要进行修改。
1. 溶液配制
（1）工作液：用实验缓冲液溶解5-Fluorouridine，配制所需浓度的工作液。
注意：最佳的工作浓度请根据实际情况调整或参阅文献自行设置梯度浓度进行摸索。工作液必须现配现用。
2. 5-Fluorouridine体内标记[1](来源文献，仅供参考)
（1）给小鼠腹膜内注射5-Fluorouridine，剂量为10μL/g，0.4M 5-Fluorouridine原液溶于0.9%盐水。
（2）注射后45min对小鼠实施安乐死，并用含有0.5% Triton X的HPEM缓冲液（2× HPEM：60mM Hepes；130mM Pipes；20mM EGTA；4mM MgCl2·6H2O）中的3.7%多聚甲醛灌注固定10min。
（3）取出其脑，用灌注缓冲液后固定1h，并在HPEM中洗涤。
（4）进行机械海马颗粒细胞解离。
（5）用小鼠单克隆抗BrdU在37°C下处理1h。
（6）用含有0.1% Tween20的PBS溶液清洗后，在室温下用FITC偶联二抗孵育细胞45min。
（7）PBS清洗后封片，在荧光显微镜下检测检测5-Fluorouridine掺入RNA的情况。
3. 5-Fluorouridine体外标记[2](来源文献，仅供参考)
（1）用2mM 5-Fluorouridine脉冲标记细胞20min。
（2）用2%甲醛固定细胞10min，用0.5% Triton X-100透化细胞10min，然后用含有0.1% Tween20的PBS溶液封闭细胞1h。
（3）用BrdU抗体在4°C下孵育过夜。
（4）PBS清洗后，在室温下用FITC偶联二抗孵育细胞1h。
（5） PBS清洗后，用DAPI处理细胞，在荧光显微镜下检测检测5-Fluorouridine掺入RNA的情况。
注意：为了您的安全和健康，请穿实验服并戴一次性手套操作。
References:
[1]Puente-Bedia A, Berciano M T, Tapia O, et al. Nuclear reorganization in hippocampal granule cell neurons from a mouse model of Down syndrome: changes in chromatin configuration, nucleoli and Cajal bodies[J]. International Journal of Molecular Sciences, 2021, 22(3): 1259.
[2]Awasthi S, Verma M, Mahesh A, et al. DDX49 is an RNA helicase that affects translation by regulating mRNA export and the levels of pre-ribosomal RNA[J]. Nucleic Acids Research, 2018, 46(12): 6304-6317.
5-Fluorouridine is a ribonucleotide metabolite of 5-Fluorouracil with anticancer activity[1]. 5-Fluorouridine can mark active transcription sites of cells in vivo and in vitro for immunodetection of nascent RNA[2, 3]. 5-Fluorouridine is an organofluorine compound belonging to pyrimidine nucleosides and their analogs. It is uridine with a fluorine substituent at the 5th position of the uracil ring and has a mutagenic effect[4].
In vitro, 5-Fluorouridine (10µM) treatment of HCT-116 cells for 24h increased the percentage of apoptotic cells, affected the expression of 1200 different genes during apoptosis, and increased the expression of macrophage inhibitory cytokine 1 (MIC-1) by up to 100-fold[5]. 5-Fluorouridine (167μM) treatment of gastric cancer cells (MKN45 and MKN28 cells) for 24-96h inhibited cell proliferation in a time-dependent manner[6].
In vivo, 5-Fluorouridine (567-1500mg/kg/day) was intraperitoneally injected into male CBA/J mice for 20 days, which induced gastrointestinal toxicity in mice[7].
References:
[1]Ghoshal K, Jacob S T. An alternative molecular mechanism of action of 5-fluorouracil, a potent anticancer drug[J]. Biochemical pharmacology, 1997, 53(11): 1569-1575.
[2]Puente-Bedia A, Berciano M T, Tapia O, et al. Nuclear reorganization in hippocampal granule cell neurons from a mouse model of Down syndrome: changes in chromatin configuration, nucleoli and Cajal bodies[J]. International Journal of Molecular Sciences, 2021, 22(3): 1259.
[3]Awasthi S, Verma M, Mahesh A, et al. DDX49 is an RNA helicase that affects translation by regulating mRNA export and the levels of pre-ribosomal RNA[J]. Nucleic Acids Research, 2018, 46(12): 6304-6317.
[4]Gmeiner W H. Chemistry of fluorinated pyrimidines in the era of personalized medicine[J]. Molecules, 2020, 25(15): 3438.
[5]Schmittgen T D, Gissel K A, Zakrajsek B A, et al. Diverse gene expression pattern during 5-fluorouridine-induced apoptosis[J]. International journal of oncology, 2005, 27(2): 297-306.
[6]Wu F, Li R T, Yang M, et al. Gelatinases-stimuli nanoparticles encapsulating 5-fluorouridine and 5-aza-2′-deoxycytidine enhance the sensitivity of gastric cancer cells to chemical therapeutics[J]. Cancer Letters, 2015, 363(1): 7-16.
[7]Houghton J A, Houghton P J, Wooten R S. Mechanism of induction of gastrointestinal toxicity in the mouse by 5-fluorouracil, 5-fluorouridine, and 5-fluoro-2′-deoxyuridine[J]. Cancer research, 1979, 39(7_Part_1): 2406-2413.
5-Fluorouridine是5-氟尿嘧啶（5-Fluorouracil）的核糖核苷酸代谢物，具有抗癌活性[1]。5-Fluorouridine可在体内和体外标记细胞的活性转录位点，用于新生RNA的免疫检测[2, 3]。5-Fluorouridine是一种有机氟化合物，属于嘧啶核苷及其类似物，是尿嘧啶环上5位带有氟取代基的尿苷，具有诱变剂的作用[4]。
在体外，5-Fluorouridine（10µM）处理HCT-116细胞24h，增加了凋亡细胞百分比，影响了细胞凋亡期间1200种不同基因的表达，使巨噬细胞抑制性细胞因子1（MIC-1）的表达增加了高达100倍[5]。5-Fluorouridine（167μM）处理胃癌细胞（MKN45和MKN28细胞）24-96h，以时间依赖性方式抑制了细胞增殖[6]。
在体内，5-Fluorouridine（567-1500mg/kg/day）通过腹腔注射处理雄性CBA/J小鼠20天，诱导了小鼠胃肠道毒性[7]。
| Cas No. | 316-46-1 | SDF | |
| 别名 | 5-氟尿嘧啶核苷 | ||
| Canonical SMILES | OC[C@@H]1[C@H]([C@H]([C@H](N2C(NC(C(F)=C2)=O)=O)O1)O)O | ||
| 分子式 | C9H11FN2O6 | 分子量 | 262.19 |
| 溶解度 | Water: 100 mg/mL (381.40 mM); DMSO: ≥ 100 mg/mL (381.40 mM) | 储存条件 | Store at 4°C, stored under nitrogen |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.814 mL | 19.0701 mL | 38.1403 mL |
| 5 mM | 0.7628 mL | 3.814 mL | 7.6281 mL |
| 10 mM | 0.3814 mL | 1.907 mL | 3.814 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Population Pharmacokinetics of Intracellular 5-Fluorouridine 5'-Triphosphate and its Relationship with Hand-and-Foot Syndrome in Patients Treated with Capecitabine
AAPS J 2021 Jan 8;23(1):23.PMID:33417061DOI:10.1208/s12248-020-00533-1.
Capecitabine is an oral pro-drug of 5-fluorouracil. Patients with solid tumours who are treated with capecitabine may develop hand-and-foot syndrome (HFS) as side effect. This might be a result of accumulation of intracellular metabolites. We characterised the pharmacokinetics (PK) of 5-Fluorouridine 5'-triphosphate (FUTP) in peripheral blood mononuclear cells (PBMCs) and assessed the relationship between exposure to capecitabine or its metabolites and the development of HFS. Plasma and intracellular capecitabine PK data and ordered categorical HFS data was available. A previously developed model describing the PK of capecitabine and metabolites was extended to describe the intracellular FUTP concentrations. Subsequently, a continuous-time Markov model was developed to describe the development of HFS during treatment with capecitabine. The influences of capecitabine and metabolite concentrations on the development of HFS were evaluated. The PK of intracellular FUTP was described by an one-compartment model with first-order elimination (ke,FUTP was 0.028 h-1 (95% confidence interval 0.022-0.039)) where the FUTP influx rate was proportional to the 5-FU plasma concentrations. The predicted individual intracellular FUTP concentration was identified as a significant predictor for the development and severity of HFS. Simulations demonstrated a clear exposure-response relationship. The intracellular FUTP concentrations were successfully described and a significant relationship between these intracellular concentrations and the development and severity of HFS was identified. This model can be used to simulate future dosing regimens and thereby optimise treatment with capecitabine.
Determination of 5-fluorocytosine, 5-fluorouracil, and 5-Fluorouridine in hospital wastewater by liquid chromatography-mass spectrometry
J Sep Sci 2020 Aug;43(15):3074-3082.PMID:32432394DOI:10.1002/jssc.202000144.
Chemotherapeutics are pharmaceutical compounds the occurrence of which in the environment is of growing concern because of the increase in treatments against cancer diseases. They can reach the aquatic ecosystems after passing through wastewater treatment plants without complete removal. One of the most frequently used chemotherapeutics is 5-fluorouracil which exhibits a strong cytostatic effect. In this paper, an analytical methodology was developed, validated, and applied to determine 5-fluorouracil, its precursor, 5-fluorocytosine, and its major active metabolite, 5-Fluorouridine, in hospital wastewater samples. Due to the expected low concentrations after dilution and interferences present in such a complex matrix, a very selective and sensitive detection method is required. Moreover, an extraction method must be implemented prior to the determination in order to purify the sample extract and preconcentrate the target analytes at micrograms per liter concentration levels. Solid-phase extraction followed by liquid chromatography with tandem mass spectrometry was the combination of choice and all included parameters were studied. Under optimized conditions for wastewater samples analysis, recoveries from 63 to 108% were obtained, while intraday and interday relative standard deviations never exceeded 20 and 25%, respectively. Limits of detection between 61 and 620 ng/L were achieved. Finally, the optimized method was applied to samples from hospital wastewater effluents.
Immobilization of 5-Fluorouridine on chitosan
Chem Biodivers 2013 Oct;10(10):1828-41.PMID:24130026DOI:10.1002/cbdv.201300025.
The 2',3'-O-levulinic acid derivative 2b of the cancerostatic 5-Fluorouridine as well as its N(3)-farnesylated nucleolipid 2d were synthesized and coupled to H2 O-soluble chitosanes of different molecular weight and at various pH values (3.5-5.5) leading to 6 and 7. In addition, the coumarine fluorophore ATTO-488 N(9)-butanoate was bound to the biopolymer by a sequential-coupling technique to afford 9 and 10. Moreover, chitosan foils were prepared, to which 2b was coupled. Their degradation by chitosanase (from Streptomyces sp. N174) was studied UV-spectrophotometrically in a Franz diffusion cell.
A comparison of 5-Fluorouridine and 5-fluorouracil in an experimental model for the treatment of vitreoretinal scarring
Curr Eye Res 1993 May;12(5):397-401.PMID:8344064DOI:10.3109/02713689309024621.
5-Fluorouridine (5-FUR), a ribonucleotide metabolite of 5-Fluorouracil (5-FU), is a more potent inhibitor of cellular proliferation and cell-mediated contraction in vitro than 5-FU. We compared the efficacy of these two drugs in a cell injection model of proliferative vitreoretinopathy using New Zealand albino rabbits. Forty-five eyes were divided into three groups and injected intravitreally with homologous fibroblasts. Eyes were examined at the time of injection and 7, 14, 21 and 28 days thereafter. By day 28, 70.5% (12 of 17) of 5-FUR treated eyes demonstrated no appreciable proliferative or tractional activity compared with 41.7% (5 of 12) of 5-FU treated eyes and 10% (1 of 10) of control eyes (p < 0.006). Medullary ray puckers developed in 29.4% (5 of 17) and 25% (3 of 12) of 5-FUR and 5-FU treated eyes respectively. No 5-FUR treated eye developed extensive tractional or combined tractional and rhegmatogenous retinal detachment compared with 33.3% (4 of 12) of 5-FU treated eyes and 80% (8 of 10) of control eyes (p < 0.001). These results suggest that 5-Fluorouridine may be more effective than 5-FU for the treatment of vitreoretinal scarring.
Synthesis and biological evaluation of 6-substituted-5-fluorouridine ProTides
Bioorg Med Chem 2018 Feb 1;26(3):551-565.PMID:29277307DOI:10.1016/j.bmc.2017.11.037.
A new family of thirteen phosphoramidate prodrugs (ProTides) of different 6-substituted-5-fluorouridine nucleoside analogues were synthesized and evaluated as potential anticancer agents. In addition, antiviral activity against Chikungunya (CHIKV) virus was evaluated using a cytopathic effect inhibition assay. Although a carboxypeptidase Y assay supported a putative mechanism of activation of ProTides built on 5-Fluorouridine with such C6-modifications, the Hint docking studies revealed a compromised substrate-activity for the Hint phosphoramidase-type enzyme that is likely responsible for phosphoramidate bioactivation through P-N bond cleavage and free nucleoside 5'-monophosphate delivery. Our observations may support and explain to some extent the poor in vitro biological activity generally demonstrated by the series of 6-substituted-5-fluorouridine phosphoramidates (ProTides) and will be of guidance for the design of novel phosphoramidate prodrugs.