Apoptozole is a small inhibitor targets HSC70 and HSP72, with the KD values of 210nM and 140nM, respectively [1]. Apoptozole promotes the instability of the lysosomal membrane and induces lysosomal membrane permeabilization (LMP) by inhibiting lysosomal HSC70 [2]. Apoptozole has been widely used in leukemia cell models to induce cell death[3].
In vitro, Apoptozole treatment for 72 hours significantly inhibited the viability of HeLa cells, SK-OV-3 cells and U373 cells, with the IC50 values of 5.92±0.47µM, 4.25±0.37µM, and 2.23±0.71µM, respectively[4]. Apoptozole treatment with 3μM for 72 hours inhibited the replication of Zika virus in Huh7 cells, reduced the production of Zika virus proteins, without toxicity to the cells[5]. Treatment with 0.5μM Apoptozole for 12h resulted in increased levels of cleaved caspase-1 and increased IL-1β abundance in NLRC4-H443p-transfected HEK293T cells[6].
In vivo, Apoptozole was injected intraperitoneally at a dose of 4mg/kg every other day for 2 weeks, which significantly inhibited the tumor volume of A549 xenograft tumor mice without affecting the body weight of the mice[7]. 30 minutes before the cecal ligation and puncture (CLP) procedure in mice, a single dose of 10mg/kg of Apoptozole was intraperitoneally injected, resulting in a significant decrease in the survival rate of mice[8].
References:
[1]Evans L E, Cheeseman M D, Yahya N, et al. Investigating apoptozole as a chemical probe for HSP70 inhibition[J]. PLoS One, 2015, 10(10): e0140006.
[2] Park S H, Ko W, Park S H, et al. Evaluation of the interaction between Bax and Hsp70 in cells by using a FRET system consisting of a fluorescent amino acid and YFP as a FRET pair[J]. ChemBioChem, 2020, 21(1-2): 59-63.
[3] Park S H, Kim W J, Li H, et al. Anti-leukemia activity of a Hsp70 inhibitor and its hybrid molecules[J]. Scientific reports, 2017, 7(1): 3537.
[4] Park S H, Baek K H, Shin I, et al. Subcellular Hsp70 inhibitors promote cancer cell death via different mechanisms[J]. Cell chemical biology, 2018, 25(10): 1242-1254. e8.
[5] Yang J, Xu Y, Yan Y, et al. Small molecule inhibitor of ATPase activity of HSP70 as a broad-spectrum inhibitor against flavivirus infections[J]. ACS infectious diseases, 2020, 6(5): 832-843.
[6] Raghawan A K, Ramaswamy R, Radha V, et al. HSC70 regulates cold-induced caspase-1 hyperactivation by an autoinflammation-causing mutant of cytoplasmic immune receptor NLRC4[J]. Proceedings of the National Academy of Sciences, 2019, 116(43): 21694-21703.
[7] Ko S K, Kim J, Na D C, et al. A small molecule inhibitor of ATPase activity of HSP70 induces apoptosis and has antitumor activities[J]. Chemistry & biology, 2015, 22(3): 391-403.
[8] Yuan X, Chen Y, Chen G, et al. The heat shock protein 70 plays a protective role in sepsis by maintenance of the endothelial permeability[J]. BioMed Research International, 2020, 2020(1): 2194090.
Apoptozole是一种小分子抑制剂，可靶向HSC70和HSP72，KD值分别为210nM和140nM[1]。Apoptozole通过抑制溶酶体HSC70，促进溶酶体膜的不稳定性并诱导溶酶体膜透化[2]。Apoptozole已广泛应用于白血病细胞模型中诱导细胞死亡[3]。
在体外，Apoptozole处理72小时能显著抑制HeLa细胞、SK-OV-3细胞和U373细胞的活力，IC50值分别为5.92±0.47µM、4.25±0.37µM和2.23±0.71µM[4]。使用3μM的Apoptozole处理Huh7细胞72小时，可抑制Zika病毒复制并减少病毒蛋白产量，且对细胞无毒性[5]。用0.5μM的Apoptozole处理转染NLRC4-H443p的HEK293T细胞12小时，会导致裂解型caspase-1水平升高和IL-1β含量增加[4]。
在体内，隔天腹腔注射4mg/kg剂量的Apoptozole，连续2周，能显著抑制A549移植瘤小鼠的肿瘤体积，且不影响小鼠体重[7]。在小鼠盲肠结扎穿刺手术前30分钟单次腹腔注射10mg/kg剂量的Apoptozole，会导致小鼠的存活率显著下降[8]。