西他列汀磷酸鹽一水合物作用機制 - Medchemexpress - MCE中國

1、Product Data SheetSitagliptin phosphate monohydrateCat. No.: HY-13749BCAS No.: 654671-77-9分式: CHFNOP分量: 523.32作靶點: Dipeptidyl Peptidase; Autophagy作通路: Metabolic Enzyme/Protease; Autophagy儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 H2O : 33 mg/mL (63.06 mM)* means

2、soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 1.9109 mL 9.5544 mL 19.1088 mL5 mM 0.3822 mL 1.9109 mL 3.8218 mL10 mM 0.1911 mL 0.9554 mL 1.9109 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲存時，請在 6 個內(nèi)使，-20C 儲存

3、時，請在 1 個內(nèi)使。BIOLOGICAL ACTIVITY物活性 Sitagliptin(MK 0431)DPP4效抑制劑，在Caco-2細胞中IC50值為19nM。IC & Target IC50: 19 nM (DPP4, in Caco-2 cell extracts)體外研究 Sitagliptin phosphate exhibits a potent inhibitory effect on DPP-4 with IC50 of 19 nM from Caco-2 cell extracts1. Sitagliptin reduces in vitro migration of

4、isolated splenic CD4 T-cells through a pathway involving cAMP/PKA/Rac1activation2. A recent study demonstrates that sitagliptin exerts a novel, direct action in order to stimulate GLP-1secretion by the intestinal L cell through a DPP-4-independent, protein kinase A- and MEK-ERK1/2-dependentpathway.

5、It therefore reduces the effect of autoimmunity on graft survival3.體內(nèi)研究In vivo, the ED50 value of sitagliptin phosphate for inhibition of plasma DPP-4 activity is calculated to be 2.3 mg/kg 7Page 1 of 2 www.MedChemEhour postdose and 30 mg/kg 24 hour postdose in freely fed Han-Wistar rats1. The strep

6、tozotocin-induced type 1diabetes mouse model exhibits elevated DPP-4 levels in the plasma that can be substantially inhibited in mice on anSitagliptin phosphate diet. This is achieved by a positive effect on the regulation of hyperglycemia, potentiallythrough prolongation of islet graft survival4. T

7、he plasma clearance and volume of distribution of Sitagliptinphosphate are higher in rats (40-48 mL/min/kg, 7-9 L/kg) than in dogs (9 mL/min/kg, 3 L/kg); and its half-life isshorter in rats,2 hours compared with 4 hours in dogs5.PROTOCOLKinase Assay 1 DPP-4 is extracted from confluent Caco-2 cells.

8、After 5 minutes of incubation at room temperature with lysis buffer(10 mM Tris-HCl, 150 mM NaCl, 0.04 U/mL aprotinin, 0.5% Nonidet P40, pH 8.0), cells are centrifuged at 35,000 g at4C for 30 minutes, and the supernatant is stored at -80C. Assays are performed by mixing 20 L of appropriatecompound di

9、lutions with 50 L of the substrate for the DPP-4 enzyme, H-Ala-Pro-7-amido-4-trifluoromethylcoumarin (final concentration in the assay, 100 M) and 30 L of the Caco-2 cell extract (diluted 1000-fold with 100 mM Tris-HCl, 100 mM NaCl, pH 7.8). Plates are incubated at room temperature for 1 hour, andfl

10、uorescence is measured at excitation/emission wavelengths of 405/535 nm using a SpectraMax GeminiXS.Dissociation kinetics of inhibitors from the DPP-4 enzyme is determined after a 1-hour preincubation of Caco-2 cellextracts with high inhibitor concentrations (30 nM for BI 1356, 3 M for vildagliptin)

11、. The enzymatic reaction is startedby adding the substrate H-Ala-Pro-7-amido-4-trifluoromethylcoumarin after a 3000-fold dilution of thepreincubation mixture with assay buffer. Under these conditions, the difference in DPP-4 activity at a certain timepoint in the presence or absence of an inhibitor

12、reflects the amount of this inhibitor still bound to the DPP-4 enzyme.Maximal reaction rates (fluorescence units/seconds 1000) at 10-minute intervals are calculated using the SoftMaxsoftware of the SpectraMax and corrected for the rate of an uninhibited reaction (vcontrol-vinhibitor)/vcontrol.MCE ha

13、s not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 CD4T-cells are plated on membrane inserts in serum-free RPMI 1640, and cell migration is assayed using Transwellchambers (Corning), in the presence or absence of purified porcine kidney DPP-4 (32.1

14、units/mg; 100 mU/mL finalconcentration) and DPP-4 inhibitor (100 M). After 1 hour, cells on the upper surface are removed mechanically, andcells that have migrated into the lower compartment are counted. The extent of migration is expressed relative to thecontrol sample.MCE has not independently con

15、firmed the accuracy of these methods. They are for reference only.Animal Mice: Overnight fasted C57BL/6J mice are challenged 45 min after compound administration with an oral glucoseAdministration 1 load (2 g/kg). Blood samples for glucose measurement are obtained by tail bleed predose and at serial

16、 time pointsafter the glucose load. To evaluate the duration of the effect on glucose tolerance, vehicle or DPP-4 inhibitors areadministered 16 h before the glucose challenge.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻 Oxid Med Cell Long

17、ev. 2019 Nov. J Biol Chem. 2018 Dec 7;293(49):18864-18878. Sci Rep. 2019 Mar 11;9(1):4074. Nutr Neurosci. 2018 Apr 26:1-17. Neurol Res. 2018 Sep;40(9):736-743.See more customer validations on HYPERLINK www.MedChemE www.MedChemEPage 2 of 3 www.MedChemEREFERENCES1. Thomas, L., et al. (R)-8-(3-amino-pi

18、peridin-1-yl)-7-but-2-ynyl-3-methyl-1-(4-methyl-quinazolin-2-ylm ethyl)-3,7-dihydro-purine-2,6-dione (BI 1356), anovel xanthine-based dipeptidyl peptidase 4 inhibitor, has a superior potency and longer duration of acti2. Kim, S.J., et al., Dipeptidyl peptidase IV inhibition with MK0431 improves islet graft survival in diabetic NOD mice partially via T-cell modulation.Diabetes, 2009. 58(3): p. 641-51.3. Sangle, G.V., et al., Novel biological action of the dipeptidylpeptidase-IV inhibitor, sitagliptin, as a glucagon-like peptide