Apoptozole (Apoptosis Activator VII) is an inhibitor of the ATPase domain of Hsc70 and Hsp70, and can induce apoptosis.

HSC70:0.21 μM(Kd), HSP70:0.14 μM(Kd)

Apoptozole通过阻断HSP70与APAF-1的相互作用来诱导依赖于caspase的凋亡，而不影响HSP70与ASK1、JNK、BAX和AIF的相互作用[2]。然而，在水性条件下，apoptozole可能形成可与HSP70蛋白非特异性相互作用的聚集体，可能导致假阳性结果和数据不一致[3]。

Apoptozole 在接受癌细胞异种移植的小鼠体内显著抑制肿瘤生长，而不影响小鼠的活力。Az 在血液中的消除半衰期 (T1/2) 显著长于 Dox（8.04 对比 1.60 小时），并且 Az 达到最大浓度 (Tmax) 所需的时间短于 Dox（1.00 对比 4.00 小时）[2]。

Inhibition of HER2/erbB2 tyrosine kinase activity: BT-474 cells are seeded on 24-well plates and cultured overnight. Mubritinib is then added at various concentrations. After incubation for 2 hours, the cells are harvested directly into sodium dodecyl sulfate (SDS)-sample buffer (200 μL). Aliquots containing equal amounts of total cell extract are run on 7.5% to 15% gradient SDS–polyacrylamide gel electrophoresis (PAGE). Following electrophoresis, proteins are transferred onto a polyvinylidene fluoride (PVDF) membrane, for western blot analysis using a relevant primary antibody. Detection of protein is accomplished by an enhanced chemiluminescent (ECL) detection method. The extent of tyrosine phosphorylation of HER2/erbB2 is measured by the LAS-1000 plus lumino-image analyser. The concentration of Mubritinib that inhibits HER2/erbB2 phosphorylation by 50% (IC50) is calculated from a dose–response curve generated by least-squares linear regression of the response using SAS software.

Several cancer cell lines (A549, lung adenocarcinoma epithelial cells; HeLa, cervical cancer cells; MDA-MB-231, breast cancer cells; HepG2, liver cancer cells) are treated with various concentrations (0-15 μM) of Az or compound 7 as a negative control for 18 hr. Cell viabilities are then determined using an MTT assay.(Only for Reference)