Sitagliptin-phosphate-MK0431-phosphate-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemESitagliptin phosphateCat. No.: HY-13749ACAS No.: 654671-78-0Synonyms: MK0431 phosphate分式: CHFNOP分量: 505.31作靶点: Dipeptidyl Peptidase; Autophagy作通路: Metabolic Enzyme/Protease; Autophagy储存式: Please store the product under the recom

2、mended conditions inthe COA.BIOLOGICAL ACTIVITY物活性 Sitagliptin(MK 0431)磷酸盐是DPP4抑制剂，对Caco-2细胞提取物的IC50为19 nM。IC50 & Target IC50: 19 nM (DPP4, in Caco-2 cell extracts)体外研究 Sitagliptin phosphate exhibits a potent inhibitory effect on DPP-4 with IC50 of 19 nM from Caco-2 cellextracts 1. Sitagliptin reduc

3、es in vitro migration of isolated splenic CD4 T-cells through a pathway involvingcAMP/PKA/Rac1 activation 2. A recent study demonstrates that sitagliptin exerts a novel, direct action inorder to stimulate GLP-1 secretion by the intestinal L cell through a DPP-4-independent, protein kinase A-and MEK-

4、ERK1/2-dependent pathway. It therefore reduces the effect of autoimmunity on graft survival 3.体内研究 In vivo, the ED50 value of sitagliptin phosphate for inhibition of plasma DPP-4 activity is calculated to be 2.3mg/kg 7 hour postdose and 30 mg/kg 24 hour postdose in freely fed Han-Wistar rats 1. The

5、streptozotocin-induced type 1 diabetes mouse model exhibits elevated DPP-4 levels in the plasma that can be substantiallyinhibited in mice on an Sitagliptin phosphate diet. This is achieved by a positive effect on the regulation ofhyperglycemia, potentially through prolongation of islet graft surviv

6、al 4. The plasma clearance and volumeof distribution of Sitagliptin phosphate are higher in rats (40-48 mL/min/kg, 7-9 L/kg) than in dogs (9mL/min/kg, 3 L/kg); and its half-life is shorter in rats,2 hours compared with 4 hours in dogs 5.PROTOCOLKinase Assay 1 DPP-4 is extracted from confluent Caco-2

7、 cells. After 5 minutes of incubation at room temperature with lysis1/3 Master of Small Molecules 您边的抑制剂师www.MedChemEbuffer (10 mM Tris-HCl, 150 mM NaCl, 0.04 U/mL aprotinin, 0.5% Nonidet P40, pH 8.0), cells are centrifugedat 35,000 g at 4C for 30 minutes, and the supernatant is stored at -80C. Assa

8、ys are performed by mixing20 L of appropriate compound dilutions 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 cellextract (diluted 1000-fold with 100 mM Tris-HCl, 100 mM NaCl, pH 7.8). Pl

9、ates are incubated at roomtemperature for 1 hour, and fluorescence is measured at excitation/emission wavelengths of 405/535 nmusing a SpectraMax GeminiXS. Dissociation kinetics of inhibitors from the DPP-4 enzyme is determined aftera 1-hour preincubation of Caco-2 cell extracts with high inhibitor

10、concentrations (30 nM for BI 1356, 3 M forvildagliptin). The enzymatic reaction is started by adding the substrate H-Ala-Pro-7-amido-4-trifluoromethylcoumarin after a 3000-fold dilution of the preincubation mixture with assay buffer. Under theseconditions, the difference in DPP-4 activity at a certa

11、in time point in the presence or absence of an inhibitorreflects the amount of this inhibitor still bound to the DPP-4 enzyme. Maximal reaction rates (fluorescenceunits/seconds 1000) at 10-minute intervals are calculated using the SoftMax software of the SpectraMaxand corrected for the rate of an un

12、inhibited reaction (vcontrol-vinhibitor)/vcontrol.MCE has 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 usingTranswell chambers (Corning), in the pre

13、sence or absence of purified porcine kidney DPP-4 (32.1 units/mg;100 mU/mL final concentration) and DPP-4 inhibitor (100 M). After 1 hour, cells on the upper surface areremoved mechanically, and cells that have migrated into the lower compartment are counted. The extent ofmigration is expressed rela

14、tive to the control sample.MCE has not independently confirmed 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 oralAdministration 1 glucose load (2 g/kg). Blood samples for glucose meas

15、urement are obtained by tail bleed predose and atserial time points after the glucose load. To evaluate the duration of the effect on glucose tolerance, vehicleor DPP-4 inhibitors are administered 16 h before the glucose challenge.MCE has not independently confirmed the accuracy of these methods. Th

16、ey are for reference only.户使本产品发表的科研献 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.MedChemEREFERENCES1. Thomas, L., et al. (R)-8-(3-amino-piperidin-1-y

17、l)-7-but-2-ynyl-3-methyl-1-(4-methyl-quinazolin-2-ylm ethyl)-3,7-dihydro-purine-2,6-dione(BI 1356), a novel 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 sur

18、vival in diabetic NOD mice partially via T-cell2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEmodulation. 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-1 secretagogue.Endocrinology, 2012. 153(2): p. 564-73.4. Kim, S.J., et al., Inhibition of dipeptidyl peptidase IV with sitagliptin (MK0431) prolongs islet graft survival in streptozotocin-induceddiabetic mice. Diabetes, 2008. 57(5): p. 1331-9.5. Beconi, M.G., et al. Disposition of the dipeptidyl peptida