Fig. 6: In vivo PK/PD profiling of KH1 (24).
From: Discovery of a CNS active GSK3 degrader using orthogonally reactive linker screening
a Pharmacokinetic profile of KH1 (24) in female Balb/c mice following a single intravenous (i.v.) bolus dose of 0.37 mg/kg (black dot) or oral (p.o.) gavage dose of 3.0 mg/kg (green cross). Mean of samples from three mice per dose route showing whole blood concentration-time profile. b Serum shift assay. Degradation of GSK3β-HiBiT by KH1 (24) in GSK3β-HiBiT KI HEK293 cells in presence of 10% foetal bovine serum (FBS) (green) or mouse serum (MS) (black) (24 h, n = 3 biological replicates, with error bars representing mean and ± SD, 4-parameter non-linear curve fitting (GraphPad)). c Immunoblotting based endogenous degradation profile of KH1 (24) in mouse embryonic fibroblast (MEF) cells for 4 h with indicated concentrations (Data representative of n = 3 biological replicates). d Dose dependent GSK3β degradation of KH1 (24) in MEF cells (4 h, n = 3 biological replicates, with error bars representing mean, ± SD, 4-parameter non-linear curve fitting (GraphPad)). e Assessment of GSK3β in liver and brain following single i.v. dose of KH1 (24) 5 mg/kg in female Balb/c mice after 4 h, data shows samples from three mice compared to control (CNT). f Relative quantification of GSK3β levels in brain and liver from (e) following a single i.v. bolus dose of  5 mg/kg of KH1 (24) (green) compared to control (CNT) (black) (n = 3 mice, error bars representing mean and ± SD, two-tailed unpaired t-test, p values calculated as p = 0.0159 for brain samples and p < 0.0001 for liver samples (GraphPad)). g Whole Proteome (green) and h Phospho-proteome (purple) analysis of liver tissue samples (n = 3 mice) collected from the pharmacodynamic study as detailed in (e). Two-tailed t-test performed with 250 randomisations to assess the statistical significance between groups using Perseus software.
