Adezmapimod (SB203580)

Systemic lupus erythematosus (SLE) are established in female MRL/lpr mice and female C57BL/6 mice

4-(4-(4-fluorophenyl)-2-(4-(methylsulfinyl)phenyl)-1H-imidazol-5-yl)pyridine

SB203580 is a p38 MAPK inhibitor with IC50 of 0.3-0.5 uM and blocks PKB phosphorylation with IC50 of 3-5 uM.

First reported p38 inhibitor

0.3–0.5 uM, 0.3–0.5 uM, 0.3–0.5 uM, 0.3–0.5 uM, 0.3–0.5 uM, 0.3–0.5 uM

SB203580 protects pig myocardium against ischemic injury in an in vivo model. [4]SB203580 is effective to prevent and treat the disease in MRL/lpr mice model of Systemic lupus erythematosus (SLE). [5]

Cellular receptor kinase phosphorylation assay, 4 ug of sheep anti-PKBalpha is immobilized on 25 uL of protein G-Sepharose overnight (or 1.5 hours) and washed in Buffer A (50 mm Tris, pH 7.5, 1 mm EDTA, 1 mm EGTA, 0.5 mm Na3VO4, 0.1% beta-mercaptoethanol, 1% Triton X-100, 50 mm sodium fluoride, 5 mm sodium pyrophosphate, 0.1 mm phenylmethylsulfonyl fluoride, 1 ug/mL aprotinin, pepstatin, leupeptin, and 1 um microcystin). The immobilized anti-PKB is then incubated with 0.5 ml of lysate (from 5 x 106 cells) for 1.5 hours and washed three times in 0.5 mL of Buffer A supplemented with 0.5 m NaCl, two times in 0.5 mL of Buffer B (50 mm Tris-HCl, pH 7.5, 0.03% (w/v) Brij-35, 0.1 mm EGTA, and 0.1% beta-mercaptoethanol), and twice with 100 ul of assay dilution buffer, 5x assay dilution buffer is 100 mm MOPS, pH 7.2, 125 mm beta-glycerophosphate, 25 mm EGTA, 5 mm sodium orthovanadate, 5 mm DTT. To the PKB enzyme immune complex is added 10 uL of assay dilution buffer, 40 um protein kinase A inhibitor peptide, 100 um PKB-specific substrate peptide, and 10 uCi of [gamma-32P]ATP, all made up in assay dilution buffer. The reaction is incubated for 20 minutes at room temperature with shaking, then samples are pulse spun, and 40 uL of reaction volume are removed into another tube to which is added 20 uL of 40% trichloroacetic acid to stop the reaction. This is mixed and incubated for 5 minutes at room temperature, and 40 uL is transferred onto P81 phosphocellulose paper and allowed to bind for 30 seconds. The P81 piece is washed three times in 0.75% phosphoric acid then in acetone at room temperature., gamma-32P incorporation is then measured by scintillation counting.

377, 43

Data from [J Biol Chem , 2010, 285, 3282432833], SB 203580purchased from Selleck, A, effects of p38 inhibitor SB203580 (1, 5, and 10 M) on SRP72 protein expression was evaluated by WB using antibodies against human SRP72, SRP54, and GAPDH. A decreased intensity of SRP72 bands was noted when using the inhibitor at 5 M concentration at 240 min (lane 6) and 10 M at 120 and 240 min (lanes 8 and 9). B, results were analyzed and RUA illustrated, finding significant results at 5 M, 240 versus 0 min, and 10 M, 240 versus 0 and 120 versus 0 min, respectively, (p0.05).

DMSO 76 mg/mL, Water <1 mg/mL, Ethanol 5 mg/mL