盐酸吉西他滨作用机制 - Medchemexpress - MCE中国

1、Product Data SheetGemcitabine hydrochlorideCat. No.: HY-B0003CAS No.: 122111-03-9分式: CHClFNO分量: 299.66作靶点: DNA/RNA Synthesis; Nucleoside Antimetabolite/Analog; Autophagy作通路: Cell Cycle/DNA Damage; Autophagy储存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解

2、性数据体外实验 H2O : 66.66 mg/mL (222.45 mM)DMSO : 46.67 mg/mL (155.74 mM; Need ultrasonic)DMF : 2.5 mg/mL (8.34 mM; Need ultrasonic)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 3.3371 mL 16.6856 mL 33.3712 mL5 mM 0.6674 mL 3.3371 mL 6.6742 mL10 mM 0.3337 mL 1.66

3、86 mL 3.3371 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 (protect from light)。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。BIOLOGICAL ACTIVITY物活性 Gemcitabine Hydrochloride (LY 188011 hydrochloride) 种 DNA合成 抑制剂，抑制 BxPC-3，Mia Paca-2，PANC- 1，PL-45 和 AsPC-1细胞长

4、的 IC50 分别为 37.6, 42.9, 92.7, 89.3 和 131.4 nM。IC & Target DNA synthesis1体外研究 MTS assay demonstrates that Gemcitabine (Hydrochloride) at 15 nM, indole-3-carbinol (I3C) at 50 M and thecombination does not affect hTERT-HPNE cell viability. However, treatment with Gemcitabine (Hydrochloride) at 15nM, I3C

5、 at 50 M and the combination results in 31%, 19% and 72% cell death of BxPC-3 cells, respectively1.体内研究The aim of study is to formulate PLGA nanoparticles (NPs) of Gemcitabine (Hydrochloride, Gemcitabine HCl) forenhanced oral bioavailability via absorption through M cells of Peyers patches. Gemcitab

6、ine HCl is available as i.v.Page 1 of 2 www.MedChemEinfusion due to its short half life (8-17 min), rapid metabolism and limited tumor uptake. Gemcitabine loaded PLGANPs shows 21.47-fold increase in relative bioavailability in comparison to plain drug solution after oral delivery2.After i.v. injecti

7、on of Gemcitabine at doses of 50, 100, and 120, and 300 mg/kg, the highest dose caused considerablebody weight loss (p0.05 at all the time points evaluated, starting from day 3 of first injection) compared with that ofthe untreated group and complete mortality, whereas 120 mg/kg is determined as the

8、 lethal dose 10% (LD10) and100 mg/kg is considered as the maximal tolerated dose, which does not cause any mortality and a minimal bodyweight loss3.PROTOCOLCell Assay 12 Cells (the human pancreatic cell lines, Mia PaCa-2, BxPC-3, AsPC-1, PANC-1, PL-45, and normal pancreatic ductalepithelial cells, h

9、TERT-HPNE cells) are seeded into 96-well plates (3000 cells/well) in triplicate. After overnightincubation, the medium is changed and cells are treated with I3C and/or NBMPR for 24 h. The medium is changedagain and cells are cultured in medium containing different concentrations of Gemcitabine in th

10、e presence orabsence of the same concentrations of I3C and/or NBMPR for 48 h. The cells are then subjected to CellTiter 96AQueous One Solution Cell Proliferation Assay (MTS). Absorbance at 490 nm is measured 2 h after the addition of 20L of MTS reagent/well1.The in vitro cytotoxicity of Gemcitabine

11、HCl loaded NPs on Caco-2 cells are performed for 6 h to check the toxicity ofNPs during the transport/permeability studies and antiproliferative effect on K562 cells is evaluated for 48 h using theMTT assay. The cells are cultured in 96-well plates at a seeding density of 1.0104 cells/well for 48 h.

12、 Experiments areinitiated by replacing the culture medium in each well with 150 L of sample solutions (0.1, 1, 10, 100 g/mL) at 37Cin the CO2 incubator. After 4, 24 and 48 h of incubation, the medium is removed and 150 L of MTT reagent (1mg/mL) in the serum-free medium is added to each well. The pla

13、tes are then incubated at 37C for another 4 h. Atthe end of the incubation period, the medium is removed and the intracellular formazan is solubilized with 150 LDMSO and quantified by reading the absorbance at 590 nm on a micro-plate multi-detection instrument, SpectraMaxM2 with Soft Max Pro. The me

14、dium treated cells serve as controls. Percentage cell viability is calculated based on theabsorbance measured relative to the absorbance of cells exposed to the negative control2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Rats2Administration

15、 23 Three groups of male Wistar rats (n=6) are subjected to single oral dose bioavailability study. The formulations areadministered orally with the aid of a syringe and infant feeding tube. The 1st group is given distilled water, the 2ndgroup is given a solution of Gemcitabine HCl in distilled wate

16、r, and the third group received Gemcitabine HCl loadedNPs at a dose of 10 mg/kg. Blood samples (0.3 mL) are drawn by retro-orbital venous plexus puncture with the aid ofcapillary tubes at 0.5, 1, 2, 4, 24, 48, 72 h post oral dose. The samples are collected in heparinised Eppendorf tubescontaining 10

17、 M tetrahydrouridine, centrifuged at 3400 rpm for 15 min and plasma is collected. To this, 200 L ofAcetonitrile is added and vortexed for 5 min followed by centrifugation at 5000 rpm for 15 min. The organic phase isseparated and evaporated under reduced pressure in a vacuum oven. The residue is diss

18、olved in mobile phase (0.15mL), vortexed for 1 min followed by centrifugation at 13,000 rpm for 5 min. Then 20 L of supernatant is injected intothe HPLC column and analyzed by HPLC.Mice3DBA/2 mice (5-6 weeks old), weighing approximately 15 to 18 g, are used for the study. The mice are provided withs

19、tandard mouse food and water ad libitum. The L1210 wt leukemia cells are maintained in vitro, and they are injectedintravenously (1105) into the mice, to develop a systemic metastatic leukemia model. The mice are divided into sixgroups of seven to eight mice each: untreated, treated with squalene na

20、noparticles, treated with 100 mg/kgGemcitabine, treated with 20 mg/kg equivalent SQgem nanoassemblies in Gemcitabine, treated with 100 mg/kgcytarabine, and treated with 100 mg/kg cytarabine every day for 5 days. After injection of leukemia cells (day 0), allgroups of mice received the treatment by i

21、.v. injection on days 1, 5, 9, and 14 (i.e., days after injecting leukemia cells),with the exception of the untreated group and the group treated with cytarabine daily by the i.v. route. The mice aremonitored regularly for weight differences and survival.Page 2 of 3 www.MedChemEMCE has not independe

22、ntly confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Nature. 2019 Oct;574(7777):264-267. Cell Res. 2020 Apr 27. Hepatology. 2019 May;69(5):1995-2012. J Mol Med (Berl). 2019 Aug;97(8):1183-1193. Am J Cancer Res. 2020 Apr 1;10(4):1182-1193.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENC