Apoptozole
(Synonyms: Apoptosis Activator VII) 目录号 : GC14411
Apoptozole是一种小分子抑制剂，可靶向HSC70和HSP72，KD值分别为210nM和140nM。
Cas No.:1054543-47-3
Sample solution is provided at 25 µL, 10mM.
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]。
| Cell experiment [1]: | |
Cell lines | HeLa cells |
Preparation Method | HeLa cells were cultured at 37°C in Dulbecco's Modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 5% CO2. Cells (5×103 cells/100μl) were seeded in 96-well plates in triplicate, cultured for 24h, and then incubated with various concentrations of Apoptozole (0, 2, 4, 6, 8, 10μM) for 72h. 20μl of MTT (5mg/ml) was added to the medium in each well, and the mixture was then incubated for 2 hours. After removing the medium containing MTT, 100μl of DMSO was added, incubated for 30min for color development, and absorbance at 570nm was measured. |
Reaction Conditions | 0, 2, 4, 6, 8, 10μM; 72h |
Applications | Apoptozole significantly inhibited the viability of HeLa cells in a dose-dependent manner. |
| Animal experiment [2]: | |
Animal models | Male nude mice |
Preparation Method | Male nude mice (4 weeks of age) were housed in a pathogen-free room with controlled temperature and humidity. Viable A549 cells (5×106) were subcutaneously injected into the flank of mice. A549 cell xenograft mice were immediately randomly divided into two groups. The first group (n=10) served as a control group and received a vehicle only. The second group (n=10) received intraperitoneal Apoptozole (4mg/kg) every other day for 2 weeks. Tumors from all mice were measured in two dimensions with a caliper every 3 days, and tumor volumes were calculated using the formula Volume =W×L2/2, where W is the width at the widest point of the tumor and L is the length perpendicular to W. |
Dosage form | 4mg/kg; every other day for 2 weeks; i.p. |
Applications | Apoptozole treatment retarded tumor growth in a xenograft mouse model without affecting mouse viability. |
References: | |
| Cas No. | 1054543-47-3 | SDF | |
| 别名 | Apoptosis Activator VII | ||
| 化学名 | 4-[[2-[3,5-bis(trifluoromethyl)phenyl]-4,5-bis(4-methoxyphenyl)-1H-imidazol-1-yl]methyl]-benzamide | ||
| Canonical SMILES | COC(C=C1)=CC=C1C2=C(C3=CC=C(OC)C=C3)N(CC4=CC=C(C(N)=O)C=C4)C(C5=CC(C(F)(F)F)=CC(C(F)(F)F)=C5)=N2 | ||
| 分子式 | C33H25F6N3O3 | 分子量 | 625.6 |
| 溶解度 | ≤10mg/ml in ethanol;50mg/ml in DMSO;50mg/ml in dimethyl formamide | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.5985 mL | 7.9923 mL | 15.9847 mL |
| 5 mM | 319.7 μL | 1.5985 mL | 3.1969 mL |
| 10 mM | 159.8 μL | 799.2 μL | 1.5985 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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