金诺芬衍生物的合成及抗肿瘤活性

doi:10.7503/cjcu20190246

摘要/Abstract

摘要：

设计合成了一类新型金诺芬衍生物, 采用核磁共振波谱(NMR)和高分辨质谱(HRMS)确认了其结构. 以目标化合物处理肿瘤细胞后, 采用四氮唑蓝盐(MTS)法检测细胞增殖情况, 用流式细胞仪检测细胞凋亡情况, 采用蛋白质免疫印迹(Western blot)法检测总的泛素化蛋白(Ub-Prs)、 K48 位链接多聚泛素化蛋白以及蛋白酶体外源性特异性底物(GFPu)的表达情况. 结果表明, 金诺芬衍生物可通过抑制蛋白酶体功能来发挥抗肿瘤效果.

关键词: 金诺芬衍生物, 蛋白酶体, 抗肿瘤活性

Abstract:

A new type of auranofin derivatives was synthesized and characterized by means of nuclear magne-tic resonance spectroscopy(NMR) and high resolution mass spectroscopy(HRMS). After treatment of tumor cells with target compounds, the effect of the target compound on cell viability was determined by MTS assay, apoptosis was detected by flow cytometry, Western blot was used to detect the effects of target compounds on total ubiquitinated protein(Ub-Prs), K48-linked polyubiquitinated protein and the expression of protease-specific substrate(GFPu) in vitro. The results showed that the auranofin derivative exerted a better antitumor effect by inhibiting the function of the proteasome.

Key words: Auranofin derivative, Proteasome, Antitumor activity

中图分类号:

O621

TrendMD:

张培全,杨倩倩,龙惠丹,陈鑫. 金诺芬衍生物的合成及抗肿瘤活性. 高等学校化学学报, 2019, 40(10): 2097.

ZHANG Peiquan,YANG Qianqian,LONG Huidan,CHEN Xin. Synthesis and Antitumor Activity of Auranofin Derivatives ^†. Chem. J. Chinese Universities, 2019, 40(10): 2097.

图/表 5

Scheme 1 Synthetic routes of the AF derivatives

Fig.1 Cell viability of malignant A562(A, D), K549(B, E) and non-cancerous(LO2)(C, F) cells treated with Auranofin and derivatives at various concentrations for 24 h(A—C) and 48 h(D—F) a. DMSO; c/(μmol·L-1), b—f. 0.25, 0.5, 1, 2, 3. Cell viability was assessed by MTS assay. Mean±SD(n=3) are representative of three independent experiments. * P<0.05, versus DM treatment alone. Aur; AF-1; AF-2; AF-3.

Fig.2 Auranofin and derivatives induces cell apoptosis by Annexin-FITC(A1—A16) and A549 cells treated with AF, AF-1, AF-2 and AF-3 at the indicated doses for 24 h and apoptosis evaluated by flow cytometry(B) (B) Mean±SD(n=3) are representative of three independent experiments.*P<0.05, versus DMSO treatment alone. a. DMSO; c/(μmol·L-1): b. 0.25; c. 0.5; d. 1.

Fig.3 A549 cells incubated in presence of DMSO(a), AF(b), AF-1(c), AF-2(d) and AF-3(e)[0, 0.5(A1—A3) and 1(B1—B3) μmol/L] for 6 h Ub-prs., K48-linked poly-ubiquitinated proteins were assessed by immunoblot with GAPDH as a loading control. (A1, B1) Ub-Prs; (A2, B2) K48; (A3, B3) GAPDH.

Fig.4 GFPu-HEK293 cells obtained by stable transfection of HEK293 cells with GFPu(a surrogate proteasome substrate) incubated in presence of AF(a), AF-1(b), AF-2(c) and AF-3(d)(0, 0.25, 0.5, and 1 μmol/L) for 6 h, and GFPu amounts were assessed by immunoblot with GAPDH as a loading control (A1—C1) Ub-Prs; (A2—C2) GFPu; (A3—C3) GAPDH. c/(μmol·L-1): (A1—A3) 0.25; (B1—B3) 0.5; (C1—C3) 1.

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