阿扎胞苷作用机制 - Medchemexpress - MCE中国

1、Product Data Sheet5-AzacytidineCat. No.: HY-10586CAS No.: 320-67-2分式: CHNO分量: 244.2作靶点: Nucleoside Antimetabolite/Analog; DNA Methyltransferase; Bacterial; Autophagy作通路: Cell Cycle/DNA Damage; Epigenetics; Anti-infection; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 mont

2、h溶解性数据体外实验 DMSO : 31 mg/mL (126.95 mM)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 4.0950 mL 20.4750 mL 40.9500 mL5 mM 0.8190 mL 4.0950 mL 8.1900 mL10 mM 0.4095 mL 2.0475 mL 4.0950 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

3、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

4、: 2.5 mg/mL (10.24 mM); Clear solution此案可获得 2.5 mg/mL (10.24 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 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 (10.24 mM); Clear solutionPage 1 of 2 w

5、ww.MedChemE此案可获得 2.5 mg/mL (10.24 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 DMSO 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 (10.24 mM); Clear solution此案可获得 2.5 mg/mL (10.24 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

6、DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 5-Azacytidine (Azacitidine; 5-AzaC; Ladakamycin)是胞苷核苷类似物，特异型抑制 DNA 甲基化。5-Azacytidine 与DNA 结合共价捕获 DNA甲噬 (autophagy)。转移酶 (DNA methyltransferases)，有益于逆转表观遗传变化。5-Azacytidine 诱导细胞IC & Target DNMT1 Nucleoside AutophagyAntimetabolite/Analog体外研究 Unmethylated C

7、pG islands associated with a variety of genes become partially or fully methylated in tumors and canbe reactivated by 5-Azacytidine1. 5-Azacytidine acts as weak inducers of erythroid differentiation of Frienderythroleukemia cells in the same concentration range where they affect DNA methyltransferas

8、e activity2. 5-Azacytidine inhibits L1210 cells with ID50 and ID90 values of 0.019 and circa 0.15 g/mL, respectively3.体内研究 TdR-3H incorporation is significantly inhibited when the animals are exposed to 5-Azacitidine (100 mg/kg, i.p.) for 2 hror longer3.PROTOCOLKinase Assay 3 A crude cell-free extra

9、ct is isolated from LI 210 cells in culture by suspension of the cells in a given volume of0.05mol/LTris-HCl buffer, pH 7.4, and sonic extraction with a Biosonik at 70% maximal output for 30 sec. Thesupernatant is collected after centrifugation at 105,000 g for 60 min (4C) in a Model L Spinco ultrac

10、entrifuge. Thefinal protein concentration of the cell-free extracts is approximately 3 mg/mL. The extracts are used as the source ofenzymes. Ribonucleotide reductase activity is measured. A unit of enzyme is defined as the amount that catalyzeddCMP synthesis at a rate of 1 mmole/hr. The assay system

11、s for the measurement of pyrimidine nucleoside (CR) anddeoxynucleoside (TdR, CdR) kinases are essentially those described by Chu and Fischer. However, reactions areterminated by heating for 2 min in a boiling water bath, and the phosphorylated derivatives are isolated according tothe method of Bach.

12、 Fifty-jul aliquots are applied to 1-inch discs of diethylaminoethyl paper, which are then placed incounting vials and eluted with 0.5 mL of 0.5 mol/LPCA. After 1 hr, 12 mL of Diotol are added, and the radioactivity isdetermined.MCE has not independently confirmed the accuracy of these methods. They

13、 are for reference only.Cell Assay 3 Twenty mL of cells (circa 1104 cells/mL) are pipetted into sterilized culture tubes with screw caps and incubated at37C overnight. The experiment is initiated by the addition of 1 mL of 5-Azacytidine (5-azaCR) or medium for a givenperiod (from 0 to 240 min) prior

14、 to the addition of 1 mL of metabolite (or medium). Cell growth is determined twice aday for 3 days by means of a Model A Coulter counter. To determine IDSO and ID90 values, 5 mL of L1210 cells(5103 cells/mL) are incubated with the drug at 37C for 3 days, and cell growth is determined.MCE has not in

15、dependently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemEAnimal For the in vivo experiments, leukemic mice (bearing circa 1103 cells/animal) are given injections i.p. with 0.2 mL ofAdministration 3 5-Azacytidine (5-azaCR) of a given concentration. Two

16、hr later, the reaction is started by injecting 0.5 mL of labeledmetabolite (TdR-3H or UR-3H, 10 /Ci/12.5 g). After 1 hr, animals (3 mice/group) are killed by cervical fracture, andthe ascites are treated with heparin, collected, pooled, and then centrifuged immediately in a Sorvall refrigeratedcentr

17、ifuge Model R2C-B at 800g for 10 min (4C).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 PLoS Pathog. 2020 Mar. Mol Oncol. 2018 Feb;12(2):180-195. Cell Death Dis. 2018 Apr 27;9(5):497. Cell Death Dis. 2018 Jan 26;9(2):129. Cell Commun Signa

18、l. 2019 Aug 14;17(1):94.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Christman JK. 5-Azacytidine and 5-aza-2-deoxycytidine as inhibitors of DNA methylation: mechanistic studies and their implications for cancer therapy.Oncogene. 2002 Aug 12;21(35):5483-95.2. Creusot F, et al. Inhibition of DNA methyltransferase and induction of Friend erythroleukemia cell differentiation by 5-azacytidineand 5-aza-2-de