Carfilzomib 是一种不可逆的蛋白酶体抑制剂和抗肿瘤剂，用于治疗难治性多发性骨髓瘤。

Carfilzomib is an irreversible proteasome inhibitor and antineoplastic agent that is used in treatment of refractory multiple myeloma.

Carfilzomib在持续或短暂处理下,有效降低多发性骨髓瘤细胞活力. Carfilzomib增加骨小梁体积,减少骨吸收,并增强非肿瘤小鼠的骨形成.Carfilzomib在体内异种移植模型中适度降低肿瘤生长.

Carfilzomib对β5亚基中的ChT-L活性具有优先的体外抑制效力,在10 nM的剂量下具有超过80％的抑制。短时间暴露于低剂量Carfilzomib导致对于β5组成型20S蛋白酶体和β5i免疫蛋白酶体亚基的优先结合特异性。在Carfilzomib刺激的ANBL -6细胞中检测caspase活性,结果显示caspase- 8和caspase -9和caspase-3活性在8小时后大幅增加和对照组8小时后的细胞相比分别增加了3.2、3.9和6.9倍。在carfilzomib处理过的细胞中,线粒体膜的完整性减少到41%(Q1 + Q2),而在对照细胞中这一比例为75％。Carfilzomib抑制多种细胞系和患者衍生的肿瘤细胞（包括多发性骨髓瘤）中的增殖,并诱导内源性和外源性凋亡信号传导途径以及c-Jun-N端激酶（JNK）的活化。

Enzyme-linked immunosorbent assay for subunit profiling of carfilzomib: ANBL-6 cells (2 × 106/well) are plated in 96-well plates and treated with Carfilzomib doses from 0.001 to 10 μM for 1 hour. Cells are then lysed (20 mM Tris-HCl, 0.5 mM EDTA), and cleared lysates are transferred to polymerase chain reaction (PCR) plates. A standard curve is generated using untreated ANBL-6 cell lysates starting at a concentration of 6 μg protein/μL. The active site probe [biotin-(CH2)4-Leu-Leu-Leu-epoxyketone; 20 μM] is added and incubated at room temperature for 1 hour. Cell lysates are then denatured by adding 1% sodium dodecyl sulfate (SDS) and heating to 100°C, followed by mixing with 20 μL per well streptavidin-sepharose high-performance beads in a 96-well multiscreen DV plate and incubated for 1 hour. These beads are then washed with enzyme-linked immunosorbent assay (ELISA) buffer (PBS, 1% bovine serum albumin, and 0.1% Tween-20), and incubated overnight at 4°C on a plate shaker with antibodies to proteasome subunits. Antibodies used included mouse monoclonal anti-β1, anti-β2, anti-β1i, and anti-β5i, goat polyclonal anti-β2i, and rabbit polyclonal anti-β5 (affinity-purified antiserum against KLH-CWIRVSSDNVADLHDKYS peptide). The beads are washed and incubated for 2 hours with horseradish peroxidase-conjugated secondary goat antirabbit, goat antimouse or rabbit antigoat antibodies. After washing, the beads are developed using the supersignal ELISA picochemiluminescence substrate. Luminescent detection is performed. Raw luminescence is converted to μg/mL by comparison with the standard curve and expressed as the % inhibition relative to vehicle control. Curve fits are generated using the following nonsigmoidal dose-response equation: Y = Bottom + (Top-Bottom)/(1 + 10?((LogEC50 ? X) × HillSlope)), where X is the logarithm of concentration, Y is the % inhibition, and EC50 is the dose showing 50% effect.

WST-1 is used to determine the effects of proteasome inhibitor Carfilzomib on cell proliferation. The inhibition of proliferation is calculated in relation to parallel control cells that receives vehicle alone. A linear spline function is used to interpolate the median inhibitory concentration (IC50) using XLfit 4 software. The degree of resistance (DOR) is calculated using the formula: DOR = IC50(resistant cells)/IC50(sensitive cells). ANBL-6 cells pulsed with 100 nM carfilzomib are washed and suspended in PBS containing 5 μg/mL of JC-1, which exhibits potential-dependent accumulation in mitochondria. Analysis of the mitochondrial membrane potential-dependent color shift from 525 to 590 nm is carried out on a FacScan, and the data are analyzed with CellQuest software.(Only for Reference)

868540-17-4

C40H57N5O7

719.91

PR-171;卡非佐米

DMSO:47 mg/mL (65.3 mM)
Ethanol:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years