Fig. 2

Wake-promoting effects of TAK-861 in WT mice, cynomolgus monkeys, and NT1 model mice during the sleep phase. (a) Time schedule of drug administration in mice during the sleep phase. TAK-861 or vehicle was administered orally to mice at ZT5, and then EEG/EMG and locomotor activity were recorded. (b) Effect of TAK-861 at 0.3, 1, and 3 mg/kg on wakefulness time in 10-min bins for 3 h (left) and total wakefulness time for 1 h (right) after administration in WT mice. Mean ± SEM; n = 8. **p < 0.01, ***p < 0.001, compared with the vehicle-treated mice, as determined by two-tailed Shirley-Williams test. (c) Effect of TAK-861 at 10 mg/kg on wakefulness time in WT mice (n = 7) and OX2R KO mice (n = 8). ***p < 0.001, compared with vehicle-treated mice, as determined by two-tailed paired t-test. (d) Effect of TAK-861 at 0.03, 0.1, and 0.3 mg/kg on wakefulness time in orexin/ataxin-3 mice. Mean ± SEM; n = 8. ***p < 0.001, compared with vehicle-treated mice, as determined by two-tailed Williams test. (e) Quantification of [³H]-EMPA binding in brain regions (arrows) in the autoradiograms of orexin/ataxin-3 mice and WT littermates (N = 6–8). The statistical analysis was performed between orexin/ataxin-3 mice and WT littermates using an analysis of variance (ANOVA) for repeated measures followed by post hoc two-tailed Student’s t-test with Bonferroni correction. ***p < 0.001. (f) Time schedule of drug administration in cynomolgus monkeys during the sleep phase. TAK-861 or vehicle was administered orally to mice at ZT12, and then EEG/EMG and locomotor activity were recorded. (g) Effects of TAK-861 on wakefulness time at 0.3 (left) and 1 mg/kg (middle) in 1-h bins for 8 h and total wakefulness time for 8 h (right) after administration in cynomolgus monkeys. Mean ± SEM; n = 8. Left and middle: *p < 0.05, **p < 0.01, ***p < 0.001, compared with the vehicle-treated mice, as determined by repeated measures analysis of variance followed by a post hoc two-tailed paired t-test, with multiplicity adjusted using the Holm method. Right: ***p < 0.001, compared with vehicle-treated cynomolgus monkeys in each group, as determined by paired t-test. AHA anterior hypothalamic, AHip amygdalohippocampal area, Ctx-Int internal layer of cortex, EEG electroencephalogram, EMG electromyogram, Hip-CA Cornu Ammonis (CA) of the hippocampus, Hip-DG dentate gyrus of the hippocampus, KO knockout, MHA medial hypothalamic area, NAc nucleus accumbens, NT1 narcolepsy type 1, Pont-nu pontine nuclei, SEM standard error of the mean, Sup-coll superior colliculus, WT wild type, ZT zeitgeber time.