LY2811376, an orally available non-peptidic β-secretase(BACE1) inhibitor (IC50: 239-249 nM), can decrease Aβ secretion (EC50: 300 nM). It has 10-fold selectivity towards BACE1 over BACE2, and more than 50-fold inhibition over other aspartic proteases including pepsin, cathepsin D, or renin.

BACE1:239 nM-249 nM, β-Amyloid:300 nM(EC50)

在APPV717F小鼠Aβ病理模型中,LY2811376（10/30/100 mg/kg）使Aβ、sAPPβ和C99出现剂量依赖性的显著减少,这些是APP被BACE1蛋白水解后最接近的分解产物.在比格犬体内,LY2811376（5 mg/kg）使血浆中的Aβ1-x减少,给药4-12 h后最大下降85%.

LY2811376浓度依赖性降低APP过表达HEK293细胞中Aβ的分泌。LY2811376浓度依赖性抑制hBACE1,包括小分子合成肽（IC50：239 nM）和较大的嵌合蛋白底物（IC50：249 nM）。在PDAPP转基因小鼠的神经元原代培养物中,LY2811376抑制Aβ分泌（EC50：100 nM）。

Determination of enzymatic ef?ciency: The stock solution for each FRET peptide substrate is prepared at 30 mM in dimethylsulfoxide (DMSO). The huBACE1:Fc muBACE1:Fc preparation is concentrated through YM10 Centricon. to a ?nal concentration of at least 7 mg/mL. The optimal enzyme concentration for each FRET peptide substrate is determined individually at 30 μM FRET peptide substrate in 50 mM ammonium acetate, pH 4.6, 1 mg/mL BSA and 1 mM Triton X-100. The enzymatic ef?ciency (kcat /Km) of either of the BACE1 orthologs toward individual FRET peptide substrates at 15, 30 and 100 μM is determined under the optimal conditions for each substrate. The progress of the reaction is monitored by measuring an increase of the emission signal at 420 nm with excitation wavelength set at 320 nm, using a GEMINI ?uorescence plate reader. Amino acid conjugated aminobenzoate is used to convert the emission signal in the relative ?uorescence units into the molar concentration of product generated in the reaction mixture. The initial phase of the timedependence curve is ?tted with a linear function whose slope is used to calculate the initial rate for huBACE1:Fc toward each peptide substrate. The kcat /Km values are calculated from the linear dependence of the initial rate on the concentration of each peptide.

The cytotoxicity in the HEK293Swe cell model is assessed using a CellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay. (Only for Reference)