Fig. 7

Novel BTK inhibitor GB7208 demonstrates greater selectivity and CNS target occupancy than ibrutinib and ameliorates SPEAE in Biozzi mice. a, b Kinases in a 357-kinase assay inhibited greater than 50% at 1 µM concentrations of ibrutinib (a) or GB7208 (b) with 1 mM ATP. Larger red circles indicate greater inhibition. Kinome trees were generated using KinMap [11], and tree illustrations were reproduced courtesy of Cell Signaling Technology, Inc. (www.cellsignal.com). c-d Target occupancy (c) and BTK inhibitor concentration (d) in brains from n = 2–3 naïve C57BL/6J mice/group dosed orally, daily, for a total of 3 days with either ibrutinib or GB7208. Tissue was collected 1 h after the final dose. Error bars are not shown for groups with n = 2 (5 and 45 mg/kg ibrutinib groups for % BTK occupancy). Data for ibrutinib and GB7208 are from separate experiments. e, f SCH/CFA-immunized Biozzi mice were treated with vehicle or 5 mg/kg GB7208 (n = 14 mice/group) daily from Day 49 (dotted line in e) through Day 79. Arrows indicate the final day of the study (Day 79). e Clinical scores, and percent body weights relative to Day 0 (relative body weight, RBW). Gray area represents the efficacy period of treatment (period starting 15 days after treatment onset, i.e., Day 64, through study termination), during which clinical scores and RBW areas under the curves were compared. f Maximum EAE scores during the efficacy period of treatment. Data in e are shown as mean + SEM, and data in c, d, and f are shown as mean ± SEM. Significance was tested using an unpaired two-tailed t test (RBW area under the curve and relative end body weight), two-tailed Mann–Whitney test (end score and maximum score), or two-tailed Mann–Whitney test followed by the Benjamini and Hochberg procedure (clinical scores on individual days, false discovery rate (FDR) = 5%, bars and asterisks above clinical score graph indicate FDR-adjusted significance level of individual days). *p < 0.050, **p < 0.010