4’-叠氮基胞嘧啶核苷作用机制 - Medchemexpress - MCE中国

1、Product Data SheetR-1479Cat. No.: HY-10444CAS No.: 478182-28-4分式: CHNO分量: 284.23作靶点: HCV; DNA/RNA Synthesis作通路: Anti-infection; Cell Cycle/DNA Damage储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 50 mg/mL (175.91 mM)* means soluble, but saturation unknown.Solv

2、entMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 3.5183 mL 17.5914 mL 35.1828 mL5 mM 0.7037 mL 3.5183 mL 7.0366 mL10 mM 0.3518 mL 1.7591 mL 3.5183 mL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；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据您的实验动物和给药式选择适当的溶解

3、案。以下溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (8.80 mM); Clear solution此案可获得 2.5 mg/mL (8.80 mM，饱和度未知) 的澄清溶液。以 1 mL

4、作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (8.80 mM); Clear solutionPage 1 of 2 www.MedChemE此案可获得 2.5 mg/mL (8.80 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DM

5、SO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (8.80 mM); Clear solution此案可获得 2.5 mg/mL (8.80 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 R-1479 (4-Azidocytidine)种核苷类似物，是特异性的 HCV

6、 RNA 依赖性 RNA 聚合酶 (RdRp) 抑制剂。R-1479 在HCV 亚基因组复制体系中抑制 HCV 复制，IC50 为 1.28 M。IC & Target IC50: 1.28 M (HCV replication)1体外研究 R-1479 (R1479) inhibits HCV RNA replication with a mean IC50 value of 1.28 M when measured as dose-dependentreduction of Renillaluciferase activity after a 72 h incubation of prol

7、iferating replicon cells. R-1479 shows no effect oncell viability or proliferation of HCV replicon or Huh-7 cells at concentrations up to 2 mM1. The most potent andnon-cytotoxic derivative is R-1479 with an IC50 of 1.28 M in the HCV replicon system. The triphosphate of R-1479 isprepared and shown to

8、 be an inhibitor of RNA synthesis mediated by NS5B (IC50=320 nM), the RNA polymeraseencoded by HCV. R-1479 displays good activity in the replicon assay with no measurable cytotoxic or cytostatic effect2.PROTOCOLKinase Assay 1 The membrane-associated, native HCV replicase complex is isolated from 220

9、9-23 HCV replicon cells and a derivedcell line carrying HCV replicon RNA with a S282T mutation in the NS5B coding sequence. The in vitro replicase assaycontain 10 L of cytoplasmic membrane fraction, 50 mM HEPES (pH 7.5), 10 mM KCl, 10 mM dithiothreitol, 5 mMMgCl2, 20 g/mL actinomycin D, 1 mM ATP, 1

10、mM GTP, 1 mM UTP, 30 Ci of -33PCTP (3000 Ci/mmol, 10 mCi/mL),1 unit/L SUPERaseIn, 10 mM creatine phosphate, and 200 g/mL creatine phosphokinase in a final volume of 25 L.Inhibition by nucleotide analogs is determined1.MCE has not independently confirmed the accuracy of these methods. They are for re

11、ference only.Cell Assay 1 The effect of R-1479 on the incorporation of tritiated thymidine into cellular DNA is measured using the 3Hthymidine incorporation scintillation proximity assay system. MTT and WST-1 assay systems are used to measurecell viability. The ATP bioluminescence assay kit HSII is

12、used to measure intracellular ATP levels1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 J Infect Dis. 2016 Sep 1;214(5):707-11. Antivir Res. 2020 Apr. Antiviral Res. 2019 Jul 27;170:104570.Page 2 of 3 www.MedChemESee more customer validati

13、ons on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Klumpp K, et al. The novel nucleoside analog R1479 (4-azidocytidine) is a potent inhibitor of NS5B-dependent RNA synthesis and hepatitis C virusreplication in cell culture. J Biol Chem. 2006 Feb 17;281(7):3793-9.2. Smith DB, et al. Design, synth

14、esis, and antiviral properties of 4-substituted ribonucleosides as inhibitors of hepatitis C virus replication: the discovery ofR1479. Bioorg Med Chem Lett. 2007 May 1;17(9):2570-6.3. Nguyen NM, et al. A randomized, double-blind placebo controlled trial of balapiravir, a polymerase inhibitor, in adult dengue patients. J Infect Dis. 2013May 1;207(9):1442-1450.McePdfHei