TAK-715 is a p38 MAPK inhibitor for p38α.

p38β:0.20 μM, p38α:7.1 nM

TAK 715抑制LPS刺激下THP-1释放TNF-alpha，IC50为48 nM。[1] TAK 715（10 μM）抑制Wnt-3a诱导的hDvl2磷酸化以及在U2OS-EFC细胞中hDvl2的变化。[2] TAK 715的酰胺NH与p38 alpha的Met109主链羰基形成氢键。TAK 715在ATP口袋中的结合位置较高，占据了疏水后口袋、腺嘌呤区域以及前口袋，并且延伸至Gly富集环的大部分长度。[3]

TAK 715（10 mg/kg，po）在小鼠中抑制LPS诱导的TNF-alpha产生，抑制率达87.6%。TAK 715在小鼠和大鼠中分别展现了18.4%及21.1%的适度生物可用性。在大鼠中，TAK 715产生的Cmax为0.19 μg/mL，AUC(0-24小时)为1.16 μg·h/mL。TAK 715（30 mg/kg，po）在大鼠佐剂诱导性关节炎（AA）模型中显著降低了次生掌跖体积，抑制率为25%。[1]

Assay of PI3K enzyme inhibition: The inhibition of PI3Kβ, PI3Kα, PI3Kγ, and PI3Kδ is evaluated in an AlphaScreen based enzyme activity assay using human recombinant enzymes. The assay measures PI3K-mediated conversion of PIP2 to PIP3. Biotinylated PIP3, a GST-tagged pleckstrin homology (PH) domain and the two AlphaScreen beads form a complex that elicits a signal upon laser excitation at 680 nm. The PIP3 formed in the enzyme reaction competes with the biotinylated PIP3 for binding to the PH domain thus reducing the signal with increasing enzyme product. The AZD6482 is dissolved in DMSO and added to 384 well plates. PBKβ, PBKα, PBKγ, or PBKδ is added in a Tris buffer (50 mM Tris pH 7.6, 0.05% CHAPS, 5 mM DTT, and 24 mM MgCl2) and allowed to preincubate with AZD6482 for 20 minutes prior to the addition of substrate solution containing PIP2 and ATP. The enzyme reaction is stopped after 20 minutes by addition of stop solution containing EDTA and biotin-PIP3, followed by addition of detection solution containing GST-grpl PH and AlphaScreen beads. Plates are left for a minimum of 5 hours in the dark prior to analysis. The final concentration of DMSO, ATP and PIP2 in the assay are, 0.8%, 4 μM, and 40 μM, respectively. IC50 values are calculated according to the equation, y = {a+[(b-a)/(l+(x/IC50)s)]}, where y = % inhibition; a = 0%; b = 100%; s = the slope of the concentration-response curve; x = AZD6482 concentration.