An inhibitor of BACE1 and PI3K
LX2343 is an inhibitor of β-site amyloid protein cleaving enzyme 1 (BACE1/β-secretase; IC50 = 11.43 μM) and PI3K (IC50 = 15.99 μM).1 It inhibits accumulation of Aβ (1-40) (Aβ40) and Aβ42 induced by streptozotocin in both HEK293-APPSW and CHO-APP cells, which express mutant and wild-type amyloid precursor protein (APP), respectively, in a concentration-dependent manner. It also inhibits STZ-induced increases in JNK and APPThr668 phosphorylation as well as soluble APPβ (sAPPβ) protein levels. LX2343 (10 mg/kg per day) reduces levels of Aβ40 and Aβ42, as well as thioflavine S staining, in the cortex and hippocampus in the APP/PS1 transgenic mouse model of Alzheimer’s disease. It also reduces cortical levels of PI3K, Akt, and mTOR phosphorylation and accumulation of the autophagy substrate p62, indicating increased autophagy. LX2343 (10 mg/kg per day) decreases path length and escape latency time in the Morris water maze in APP/PS1 mice.
1.Guo, X.D., Sun, G.L., Zhou, T.T., et al.Small molecule LX2343 ameliorates cognitive deficits in AD model mice by targeting both amyloid β production and clearanceActa Pharmacol. Sin.37(10)1281-1297(2016)
Kinase experiment: | Inhibition of BACE1 enzyme by LX2343 is assayed using BACE1 activity kits in vitro. Briefly, BACE1 substrate (250 nM), BACE1 enzyme (0.35 U/mL), and varied concentrations of LX2343 (5, 10, and 20 μM) are sequentially incubated for 1 h at 37°C in the dark. Fluorescence intensity is measured with excitation and emission wavelengths at 545 and 585 nm, respectively[1]. |
Cell experiment: | SH-SY5Y cells are transfected with mRFP-GFP-LC3 plasmids via an adenovirus. The cells are treated without or with Streptozotocin (0.8 mM) in combination with 5 or 20 μM LX2343 for 4 h and then fixed with 4% paraformaldehyde and observed using an Olympus Fluoview FV1000 confocal microscope[1]. |
Animal experiment: | Mice[1]APP/PS1 [B6C3-Tg(APPswe, PS1dE9)] transgenic mice are used. Genotyping to confirm APP/PS1 DNA sequences in their offspring is performed by assaying the DNA from tail biopsies, with Tg-negative mice as a negative control. Twenty male APP/PS1 mice are divided into two groups with ten non-transgenic mice in one group to serve as a negative control. The two 6-month transgenic groups are administered 10 mg/kg per day of LX2343 or vehicle, and the 6-month non-transgenic group is administered the vehicle for 100 d via intraperitoneal injection. After 100 d of administration, MWM assays are applied to evaluate the cognitive abilities of the mice for 8 d under continuous LX2343 treatment. Upon completion of the MWM test, the mice are euthanized, and the brains are removed and bisected at the mid-sagittal plane. The right hemispheres are frozen and stored at -80°C, and the left hemispheres are fixed in 4% paraformaldehyde[1]. |
参考文献: [1]. Guo XD, et al. Small molecule LX2343 ameliorates cognitive deficits in AD model mice by targeting both amyloid β production and clearance. Acta Pharmacol Sin. 2016 Sep;37(10):1281-1297. | |
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