(E)-Daporinad (FK866) is a highly specific, non-competitive small molecule inhibitor of nicotinamide phosphoribosyltransferase (NAMPT) with potential anti-tumor and anti-angiogenic activities with an IC50 value of 0.09 nM.

NMPRTase:0.4 nM (Ki), NAMPT:0.09 nM

方法：用 (E)-Daporinad（ FK866 ）（100 nM，30 分钟） 预处理肝细胞，然后进行 GaIN/LPS（G/L，1 mg/mL /30 ng/mL，24 小时）攻击，观察(E)-Daporinad（ FK866 ）是否能减轻GaIN/LPS诱导的肝毒性。
结果：用(E)-Daporinad（ FK866 ）预处理的原代肝细胞在GaIN/LPS损伤后表现出自噬活性显着增加，(E)-Daporinad（ FK866 ）可以改善GaIN/LPS和ConA诱导的肝损伤，其保护机制可能涉及其通过抑制JNK诱导自噬的能力。[1]

方法：用GaIN/LPS和ConA处理小鼠前24、12和0.5小时施用(E)-Daporinad（ FK866 ）（10mg / kg，腹腔注射），测试(E)-Daporinad（ FK866 ）对小鼠ALF的潜在影响。
结果：(E)-Daporinad（ FK866 ）治疗后降低了GaIN/LPS或ConA治疗小鼠的死亡率，(E)-Daporinad（ FK866 ）处理前和处理后均能减弱小鼠GaIN/LPS和ConA诱导的ALF，且(E)-Daporinad（ FK866 ）预处理对GaIN/LPS或ConA攻击的响应效果更好。[1]

High Throughput Screening: FITC-MBM1 at 15 nM and menin at 150 nM in the FP buffer are mixed and incubated for 1h in the dark at room temperature. For point screening, the 0.2 μL of each compound (20 μM final concentration, 1% DMSO) is added to 20 μL of the aliquot of the protein-peptide mixture and incubated on 384-well plates in the dark at room temperature for 1h. In confirmation screening, the serial dilution plates with compounds in DMSO are prepared and used to titrate the menin-FITC-MBM1 complex. Change in fluorescence polarization is monitored at 525 nm after excitations at 495 nm using the PHERAstar microplate reader (BMG) and applied to determine IC50 values with the Origin 7.0 program.

For MTT assays, 0.5 × 106 cells/mL is plated in triplicate on 96-well plates. APO866 (0.01 nM-100 nM) is added in 50 μL of culture medium, with culture medium alone serving as control. After 72 or 96 hours of incubation, 15 μL of dye solution is added to each well and cells are incubated for an additional 4 hours. Stop solution (100 μL/well) is added for 1 hour and the absorbance is read at 570 nm on a spectrophotometer. For trypan blue dye exclusion staining, 0.5 × 105 cells/well is grown in 6-well plates with 1 mL media in the absence or presence of APO866 for 96 hours. Cells from each sample are incubated with 10 μL trypan blue solution (at a 1:1 ratio [vol/vol] for 1 minute). Cell survival is determined by calculating proportion of live (unstained) cells. (Only for Reference)