A-1210477 is an effective and specific MCL-1 and Bcl-2 inhibitor (Ki/IC50: 0.454/26.2 nM).

MCL1:26.2 nM

在H929细胞中，A-1210477高亲和力结合于MCL-1，并促使MCL-1蛋白表达升高。在H929、H2110和H23细胞中，A-1210477引起凋亡的典型特征，并抑制依赖MCL-1的细胞生存能力。此外，A-1210477与navitoclax协同作用，有效杀灭多种癌症细胞系。[1] 在SKBR3细胞中，A-1210477抑制MCL-1–BIM相互作用，并诱导经典凋亡特征。[2] 另外，A-1210477还使非霍奇金淋巴瘤细胞系对venetoclax (ABT-199)更为敏感。[3]

Binding affinity assays: TR-FRET-binding affinity assays are performed for BCL-2, BCL-XL, and MCL-1 in 4.52?mM monobasic potassium phosphate, 15.48?mM dibasic potassium phosphate, 1?mM sodium EDTA, 0.05% Pluronic F-68 detergent, 50?mM sodium chloride, and 1?mM DTT (pH 7.5). For MCL-1 assays, GST-tagged MCL-1 (1?nM) is mixed with 100?nM f-Bak, 1?nM Tb-labeled anti-GST antibody, and compound at room temperature (RT) for 60?min. Fluorescence is measured on an Envision plate reader using a 340/35?nm excitation filter and 520/525 (f-Bak) and 495/510?nm (Tb-labeled anti-GST antibody) emission filters.

Adherent cell lines are seeded at 50?000 cells per well in 96-well plates and treated for 48?h with compounds diluted in half-log steps starting at 30?μM and ending at 0.001?μM. Multiple myeloma cell lines were seeded at 15?000–20?000 cells per well and treated similarly. Effects on proliferation and viability were determined using CellTiter-Glo reagent according to the manufacturer's instructions. IC50 values are determined by non-linear regression analysis of the concentration response data.(Only for Reference)