Target occupancy and engagement assays show that MAGLi 432 potently reduces MAGL activity in a mouse model of LPS-induced neuroinflammation.

(A) Treatment groups and injection schedule of LPS and MAGLi 432. CD-1 mice were challenged with LPS or NaCl followed by MAGLi 432 or vehicle (i.p. administration over 3 consecutive days). Blood and brain tissue were collected for analysis. (B, C) ABPP and western blot analysis of cortical brain lysates from each treatment group reveal that MAGLi 432 achieves an effective blockade in the brain (n = 6). (C) The average signal intensities of MAGL activity and total protein were quantified as the total detectable active MAGL signal (ABPP) divided by the total MAGL/β-actin signal. (D) LC-MS analysis of 2-AG, arachidonic acid, PGE2, PGD2 and AEA in cortical lysates of each treatment group (n = 3). Data are means ± SD, one-way ANOVA with Tukey’s post hoc test, ns = not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).