PD98059 - MEK 抑制剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPD98059Cat. No.: HY-12028CAS No.: 167869-21-8分式: CHNO分量: 267.28作靶点: MEK; Autophagy作通路: MAPK/ERK Pathway; Autophagy储存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect fromlight)溶解性数据体外实验 DMSO : 16 mg/

2、mL (59.86 mM; Need ultrasonic and warming)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (9.35 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 (9.35 mM); Clear solutionBIOLOGICAL ACTIVITY1/3 Master of Small Molecules 您边的抑制剂师www.MedChemE物活性 PD98

3、059是有效，选择性和可渗透细胞的 MEK1 和 MEK2 抑制剂，IC50 分别为4 M，50 M。IC50 & Target MEK Autophagy5 M (IC50)体外研究 Concentrations of PD98059 of 20 M are not cytotoxic to cultured MCF10A, MCF10A-Neo, and MCF10A-NeoT cells. However, PD98059 is weakly cytostatic to all three lines at concentrations of 10 M. Treatment ofMCF1

4、0A-Neo and MCF10A-NeoT cultures with concentrations of PD98059 up to 20 M for 2-22 hr does notalter the total ERK content. However, treatment with PD98059 does result in concentration-dependentreductions in the dually phosphorylated forms of ERK1 and ERK2. Within 2 hr of a 10-M treatment,phosphoryla

5、ted ERK contents are reduced 74% and 86% in MCF10A-Neo and MCF10A-NeoT cultures,respectively (IC50=1 M). Within 22 hr of treatment, phosphorylated ERK forms are almost completelyeliminated in both cell lines 1. PD98059 (PD 098059) prevents the activation of MAPKK1 by Raf or MEKkinase in vitro at con

6、centrations (IC50=2-7 M). PD98059 inhibits both the activation and phosphorylation ofMAPKK1 in vitro by either c-Raf or MEK kinase with IC50 values of 42 M. Incubation of Swiss 3T3 cells withPD98059 (50 M) suppressed by 80-90% the activation of MAPKK induced by each agonist, but the activationof c-R

7、af is enhanced 2-3-fold 2.体内研究 The treatment of mice with PD98059 significantly reduces the level of p-ERK1/2. Moreover, a significantincrease in the phospho-p38 expression is observed in Zymosan-treated mice at 18 h after Zymosanadministration compared to the sham-operated mice. The treatment with

8、PD98059 significantly reduces thep38 expression 3. Repeated treatment with PD98059 attenuates mechanical allodynia measured by the vonFrey test three (18.0 g0.8, n=10) and seven (20.21 g0.67, n=26) days after CCI in comparison to thevehicle-treated CCI-exposed rats (15.1 g1.3, n=7 and 14.21 g0.44, n

9、=28, respectively). Repeated injectionof PD98059 diminishes thermal hyperalgesia, as is evaluated by the cold plate test, three (17.5 s2.1, n=10)and seven (25.54 s1.03, n=26) days following CCI compared to vehicle-treated CCI-exposed rats (11.5 s1.8,n=7 and 11.4 s0.88, n=28, respectively) 4.PROTOCOL

10、Kinase Assay 1 Kinase reactions are performed in 50 L reaction volumes and contain 50 mM Tris, pH 7.4, 10 mM MgCl2, 2mM EGTA, 10 M ATP (containing 1 Ci of 3000 Ci/mmol -32PATP), 7.6 g of GST-MEK1, 7.2 g of GST-ERK1, and 20 g of MBP. PD98059 and other flavonoids are added to the reactions mixtures im

11、mediatelyafter the addition of GST-MEK1 but before the addition of GST-ERK1 and ATP. Control reactions containERK1 and MBP but no MEK. Reaction mixtures are incubated at 30C for 15 min before being stopped bythe addition of Laemmlis SDS sample buffer. Proteins are separated on SDS-15% polyacrylamide

12、 gels. Aftervacuum drying of the gel, radioactivity is detected by autoradiography on X-ray film or phosphoimaging usinga BioRad GS-525 Molecular Imager 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 The MCF10A, MCF10A-Neo, and MCF10A-Ne

13、oT cell lines are used. Subconfluent cultures are treated withPD98059 (0-100 M). Viability of cells after treatment is assessed by ability to exclude trypan blue. Culturesearmarked for RNA isolation are washed twice with phosphate-buffered saline (2.7 mM KCl, 1.5 mM2/3 Master of Small Molecules 您边的抑

14、制剂师www.MedChemEKH2PO4, 137 mM NaCl, 8 mM Na2HPO4, pH 7.2) at harvesting and stored at -80C 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 3Administration 34 Male CD mice (20-22 g) are randomly allocated into the following groups: 1. Zymos

15、an+DMSO group. Miceare treated intraperitoneally (i.p.) with Zymosan (500 mg/kg, suspended in saline solution) and with thevehicle for PD98059 (10% DMSO, v/v) i.p. 1 and 6 h after Zymosan administration (N=10). 2. PD98059group. Identical to the Zymosan+DMSO group but are administered PD98059 (10 mg/

16、kg, i.p. bolus) at 1 and6 h after Zymosan (N=10) instead of DMSO. 3. Sham+DMSO group. Identical to the Zymosan+DMSO groupbut are administered saline solution instead of Zymosan (N=10). 4. Sham+PD98059 group. Identical toSham+DMSO group, except for the administration of PD98059 (10 mg/kg i.p. bolus)

17、1 and 6 h after salineadministration (N=10).Rats 4The rats (male Wistar, 300-350 g) are used. The PD98059 (2.5 g/5 L, i.t.) is single or repeatedpreemptively administered 16 h and 1 h before CCI and then once daily for 7 days. The Vehicle-treated CCI-exposed rats receive 75% DMSO according to the sa

18、me schedule. There is no significant difference in painbehavior between no-treated and V(DMSO)-treated CCI-exposed rats. This method of PD98059 or vehicleadministration is used throughout the study and is referred to in the text as “repeated administration”. At day7th after CCI 30 min after PD98059

19、administration tactile allodynia is measured using von Frey test andthermal hyperalgesia is conducted using cold plate test.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 Nat Immunol. 2018 Mar;19(3):233-245. Sci Transl Med. 2019 Feb 6;11(47

20、8). pii: eaau5266. J Pineal Res. 2019 Apr;66(3):e12552. Mol Cell. 2018 Feb 1;69(3):480-492.e7. Biomaterials. 2017 Jun;130:14-27.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Reiners JJ Jr, et al. PD98059 is an equipotent antagonist of the aryl hydrocarbon receptor and inhibitor of mitogen-activated proteinkinase kinase. Mol Pharmacol. 1998 Mar;53(3):438-45.2. Alessi DR, et al. PD 098059 is a specific inhibitor of the activation of mitogen-activ