PS48 - PDK-1 Activator - 生命科学试剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPS48Cat. No.: HY-15967CAS No.: 1180676-32-7分式: CHClO分量: 286.75作靶点: PDK-1作通路: PI3K/Akt/mTOR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 : 100 mg/mL (348.74 mM; Need ultrasonic)Mass Solvent1

2、 mg 5 mg 10 mg Concentration制备储备液1 mM 3.4874 mL 17.4368 mL 34.8736 mL5 mM 0.6975 mL 3.4874 mL 6.9747 mL10 mM 0.3487 mL 1.7437 mL 3.4874 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，并请注意储备液的保存式和期限。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，配制前请先配制澄清的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制

3、终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (8.72 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (8.72 mM); Clear solution3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (8.72 mM); Clear solution1/3 Maste

4、r of Small Molecules 您边的抑制剂师www.MedChemEBIOLOGICAL ACTIVITY物活性 PS48种 PDK1 激活剂，AC50 为 8 M。IC50 & Target AC50: 8 M (PDK1) 1体外研究 PS48 activates full length PDK1 (His-PDK1- FL) and PDK1 50-359Tyr288Gly;Gln292Ala (His-PDK1 dm)with AC50s (the concentrations required to reach 50% of the maximal activation)

5、 of 7.950.2 and 10.02.0 M, respectively. PS48 binds to PDK150359 with a 1:1 stoichiometry and a binding affinity in the micromolarrange (Kd=10.3 M) 1. PDK1 activator, PS48, has the ability to reverse the cell proliferation inhibition role oftriptolide (TP) in vitro. The inhibition role of TP in cell

6、 number is significantly reversed by PS48. TPsignificantly increases the cell proportion in G0-G1 phase and decreases the cell proportion in G2-M and Sphase. However, the effect of TP on cell cycle distribution is all reversed by PS48. In addition, suppression ofPDK1/Akt/mTOR pathway by TP in high g

7、lucose (HG)-treated human renal mesangial cells (HRMCs) is alsoreversed by PS48, as well as the expression of Ki-67 and proliferating cell nuclear antigen (PCNA) 2.PROTOCOLKinase Assay 1 PDK1 activity tests are performed using T308tide as a substrate for PDK1. In brief, PDK1 activity assay isperform

8、ed at room temperature (22C) in a 20 L mix containing 50 mM Tris pH 7.5, 0.05 mg/mL BSA, 0.1%-mercaptoethanol, 10 mM MgCl2, 100 M 32PATP (5-50 cpm/pmol) , 0.003% Brij, 150-500 ng PDK1, andT308tide (from 0.1 to 1 mM). When appropriate, the PDK1 activity assay is performed in a 96 well format and4 L a

9、liquots spotted on p81 phosphocellulose papers using ep motion 5070, washed in 0.01% phosphoricacid, dried, and then exposed and analysed using PhosphoImager technology. Activity measurements areperformed in duplicates or triplicates with less than 10% difference between replicates. Experiments arer

10、epeated at least twice 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 Human renal mesangial cells (HRMCs) are cultured with 1640 media, containing 10% fetal bovin serum at 37C in 5% CO2. Cells are cultured with D-glucose at normal (5.5 m

11、M) or high (25 mM) concentrations inserum-free medium. D-Mannitol (25 mM) is used for a control of osmolality. TP is reconstituted in 0.01%DMSO and freshly diluted with culture medium to 10 ug/L before using. To determine the specific role ofPDK1 in TP-potentiated anti-proliferation, 5 M PS48 (MedCh

12、em Express, USA) is applied following thetreatment of TP. MTT assay is used to detect cell proliferation. HRMCs are seeded at a density of 1x105/mLinto 96-well plates. After 12, 24, 48 and 72 h incubation with different compounds, 20 uL MTT (5 mg/mL) isadded to each well. Cells are then cultured for

13、 an additional 2 h and subsequently lysed using DMSO (150uL/well). When the formazan crystals completely dissolve, the optical density (OD) is measured at 570 nm.The arithmetic mean OD of six wells for each group is calculated 2.MCE has not independently confirmed the accuracy of these methods. They

14、 are for reference only.户使本产品发表的科研献2/3 Master of Small Molecules 您边的抑制剂师www.MedChemE Int J Biol Sci. 2017 Sep 21;13(10):1266-1275.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Hindie V, et al. Structure and allosteric effects of low-molecular-weight activators on the protein kinase PDK1. Nat Chem Biol. 2009Oct;5(10):758-64.2. Han F, et al. Triptolide Suppresses Glomerular Mesangial Cell Proliferation in Diabetic Nephropathy Is Associated with Inhibition ofPDK1/Akt/mTOR Pathway. Int J Biol Sci. 2017 Sep