生物活性

SB202190 (FHPI)is a highly selective, cell-permeable, ATP competitive inhibitor of p38α/βwith IC50 of 50 nM/100 nM. SB 202190 is a pyridinyl imidazole that inhibits the p38 pathway. A highly selective, potent and cell-permeable inhibitor of p38 MAP kinase. The compound inhibits the isoforms p38α and p38β. SB 202190 has displayed the ability to activate CPP32-like caspases and induce apoptosis. Research demonstrates that when one residue of ERK2, a related p38 not inhibited by pyridinyl imidazoles, is changed it allows SB 202190 to bind. Studies have revealed that SB 202190 can improve leukemia cell growth by activating MEK/MAPK, as well as phosphorylate and activate MLK3. Binds within the ATP pocket of the active kinase (Kd = 38 nM, as measured in recombinant human p38), and selectively inhibits the p38α and β isoforms (IC50 = 50 and 100 nM at SAPK2a/p38 and SAPK2b/p38β2 respectively).

Kinase activities of SB 202190

IC50value of SB203580 on human breast cancer cell line MDA-MB-231 is 46.6μM.[3]

SB202190inhibits CK1-mediated phosphorylation of a 6X histidine- tagged CREB fusionprotein with an IC50 of 0.6 μM.[4]

SB202190inhibits IL-1 in monocytes with an IC50 of 50nM.[2]

SB202190 inhibits CSBP binding with an IC50 of 50nM.[2]

体外研究

体内研究

1% DMSO+30% polyethylene glycol+1% Tween 80

激酶实验

Protein Kinase Assays[5]

MAPKAPK 2 assays were performed. Briefly,Jurkat cells were serum-starved for24 h and then incubated with or without thespecific p38 inhibitor SB202190 for 30 min prior to treatment with anti-Fas mAb(100ng/ml) for 2 h or left alone as indicated in the figure legends. The cellswere harvested in lysis buffer and clarified by centrifugation. Endogenous MAPKAPK2was immunoprecipitated with anti-MAPKAPK 2 polyclonal antibody for 3 h at 4 °C.The activity of the immune complex was assayed at 30 °C for 30 min in 30 ml of kinasebuffer in the presence of 1μM ATP/10μCi [γ-³²P]ATP (10Ci/mmol) with GST-hsp27 as a substrate. Thereactions were terminated with Laemmli sample buffer. The proteins wereresolved by 13% SDS-polyacrylamide gel electrophoresis followed byautoradiography. The phosphorylated proteins were quantitated by aPhosphorImager.

Mammalian CSBP2 was expressed with a (His)₆ tag in a HOG-1 deleted mutant of S. cerevisiae and activated in vivo by osmoticshock. The enzyme was purified to >95% using Nickel chelate resin. Thekinase activity of purified CSBP2 was determined by measuring the incorporationof ³²p from y-[³²p] ATP into an EGFR-derived peptide(T669) having the following sequence: KRELVEPLTPSGEAPNQALLR. Reactions (30μL)contained 25mM Hepes buffer, pH 7.4, 8mM MgCl₂;

10μM Na-vanadate; 1mM EDTA, 0.8μCi/170μM ³²P/ATP;20ng purified CSBP2; and 0.4mM peptide. Compounds were incubated for 20 min at4°C with enzyme and peptide prior to adding ATP. Reactions were incubated for10min at 30°C and were stopped by adding 10μL of 0.3 Mphosphoric acid. Phosphorylated peptide was isolated from the reaction mixtureon phosphocellulose filter paper (p81). Filters were washed with 75mMphosphoric acid and counted using a liquid scintillation counter.

细胞实验

Cell culture maintenance[3]

MDA-MB-231 human breast cancer cell linewas cultured in RPMI-1640 containing 4.5g/L glucose, 10mM HEPES, 1mM sodiumpyruvate, 0.15% sodium bicarbonate, 100μg/mlstreptomycin and100 U/ml penicillin. The medium was supplemented with 10% fetalbovine serum (FBS). The cell line was grown in 25 cm² corning flasksfor passages. For the experiments, the cells were grown in 35 x 10 mm Corningdishes, 60 x 15 mm Corning dishes or E-Plate L8. The cells were subculturedupon reaching about 70% confluence.

MTT-basedcytotoxicity assay

MDA-MB-231 cells (100,000) were seeded ineach 35 x 10 mm dish. Cells were treated at the logarithmic phase of the growthwith various doses of SB203580 (0.1, 0.5, 1, 5, 10, 25, 50, 100μM)or SB202190 (0.1, 0.5, 1, 5, 10, 25, 50, 100μM) for 24 h.Following the inhibitor exposure period, the medium was changed with RPMI-1640containing 0.5% FBS + 0.5 mg/ml MTT, the cells were incubated for another 4 hat 37^oC with 5% CO₂. After MTT treatment, cells werelysed with 200μl 3%SDS plus 1 ml 40 mM HCl/isopropanol for 15 min.  After dissolving the MTT-formazan crystals,the cell lysate was diluted with 3% SDS + 40 mM HCl/isopropanol. Finally, eachsample absorbance was recorded at 570 nm with a SmartSpec Plusspectrophotometer. The data were analyzed and graphed with GraphPad Prism 5program. The IC50 values were then obtained by

GraphPad Prism 5 program using nonlinearregression to fit the data to the log(inhibitor) versus normalized response.

动物实验

In vivo experiments[6]

Xenografts

Female athymic nu/nu mice aged 6 to 8 weekswere use. About 3 x 10⁶ tumor cells were injected subcutaneously (s.c.)into the left flank of each mouse. Tumors were detected by palpation andmeasured periodically with calipers. Mice were euthanized when the tumor volumereached 1,000 mm³.

Irinotecanand SB202190 treatment

Irinotecan stock solution was diluted in0.9% sodium chloride and 40 mg/kg were administered intraperitoneally (i.p.) totumor-bearing mice according to the following schedule: 4 injections (1 every 4days) starting when tumors reached 100 mm3. Mice in the control group received0.2 mL of 0.9% sodium chloride solution according to the same schedule. SB202190stock solution was diluted in 0.9% sodium chloride and administered i.p. at0.05mmol/kg daily for 12 days when tumors reached 100 mm3. Irinotecan +SB202190 were administered when tumors reached 100mm³: 4 injections(1 every 4 days) of irinotecan at 40mg/kg + SB202190 at 0.05mmol/kg for 12days.

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

例如，已知某工具药用于小鼠的剂量为88 mg/kg , 则用于大鼠的剂量换算方法：将88 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到该药物用于大鼠的等效剂量44 mg/kg。

参考文献

[1] Davies SP RH, Caivano M, Cohen P. Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J. 2000;351(Pt 1):95-105.
[2] Gallagher TF SG, Kassis S, Laydon JT, Blumenthal MJ, Lee JC, Lee D, Boehm JC, Fier-Thompson SM, Abt JW, Soreson ME, Smietana JM, Hall RF, Garigipati RS, Bender PE, Erhard KF, Krog AJ, Hofmann GA, Sheldrake PL, McDonnell PC, Kumar S, Young PR, Adams JL. Regulation of stress-induced cytokine production by pyridinylimidazoles; inhibition of CSBP kinase. Bioorg Med Chem. 1997;5(1):49-64.
[more]

体内溶解度
约30 mg/mL

Clinical Trial Information ( data from http://clinicaltrials.gov )

注：以上所有数据均来自公开文献，并不保证对所有实验均有效，数据仅供参考。

相关化合物库

使用AMQUAR产品发表文献后请联系我们