新型樟脑磺胺基肟醚衍生物的合成及抗肿瘤活性研究

doi:10.6023/cjoc202009050

摘要/Abstract

利用药物设计中的活性拼接原理, 以(+)-10-樟脑磺酸为原料, 经酰氯化、酰胺化和缩合反应, 设计合成了一系列樟脑磺胺基肟醚类衍生物, 通过¹H NMR、¹³C NMR和HRMS对其结构进行了表征. 采用噻唑蓝(MTT)法对目标化合物对人肺腺癌细胞(A549)、人宫颈癌细胞(Hela)、人乳腺癌细胞(MCF-7)和人正常胚胎肝细胞(LO2)进行抗肿瘤活性评价。结果表明, 大部分化合物显示出良好的抗肿瘤活性。其中, (+)-1-(7,7-二甲基-2-(苄氧基亚氨基)双环[2.2.1]庚烷-1-基)-N-(3-(三氟甲基)苯基)甲磺酰胺(4r)对三种细胞表现出最好的抗增值效果, IC₅₀值分别为6.75、7.93和4.51 μmol?L ^–1. 采用流式细胞术检测细胞周期和凋亡, Hoechst染色观察细胞形态变化. 荧光探针DCFH-DA和JC-1分别用于检测细胞内活性氧水平和线粒体膜电位. 初步机理研究表明, 化合物4r可将MCF-7细胞阻滞在G0/G1期, 可诱导活性氧产生和线粒体膜电位崩溃进而诱导细胞呈剂量依赖式凋亡.

关键词: 樟脑磺胺基肟醚, 抗肿瘤活性, G0/G1期阻滞, 凋亡

Based on the combination principle in drug design, a series of camphor sulfamoxime ether derivatives were designed and synthesized with (+)-10-camphorsulfonic acid as the staring materials via acyl chlorination, acyl amidation and condensation. The target compounds were characterized by ¹H NMR, ¹³C NMR and HRMS spectra. In addition, the antiproliferative effects of compounds were evaluated on a panel of human adherent cell lines (A549, Hela, MCF-7 and LO2) by means of methyl thiazolyl tetrazolium (MTT) assays. Noticeably, (+)-1-(2-((benzyloxy)imino)-7,7-dimethylbicyclo[2.2.1]-heptan- 1-yl)-N-(3-(trifluoromethyl)phenyl)methanesulfonamide (4r) possessed excellent antiproliferative against A549 (6.75 μmol? L ^–1), Hela (7.93 μmol?L ^–1) and MCF-7 (4.51 μmol?L ^–1). Flow cytometry was used to detect cell cycle progression and apoptosis, and morphological change of apoptosis was observed by Hoechst 33258 staining. Levels of reactive oxygen specy (ROS) and mitochondrial membrane potential were measured by DCFH-DA and JC-1, respectively. These results indicated that compound 4r could cause cell cycle arrest at G0/G1 phase and triggered apoptosis in MCF-7 cells. Furthermore, 4r could induce the accumulation of ROS and mitochondrial disruption. Taken together, compound 4r could be a potential anticancer drug candidate.

Key words: camphor sulfamoxime ether, antitumor activity, G0/G1 phase arrest, apoptosis

引用此文

赵雨珣, 王芸芸, 张成龙, 徐徐, 王石发. 新型樟脑磺胺基肟醚衍生物的合成及抗肿瘤活性研究[J]. 有机化学, 2021, 41(3): 1224-1233.

Yuxun Zhao, Yunyun Wang, Chenglong Zhang, Xu Xu, Shifa Wang. Synthesis of Novel Camphor Sulfamoxime Ether Derivatives and Its Application in Antitumor Activity[J]. Chinese Journal of Organic Chemistry, 2021, 41(3): 1224-1233.

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图/表 7

Figure 1. Chemical structures of oxime ethers and their derivatives

Scheme 1. Synthesis of compounds 4a~4r

Compd.	IC₅₀/(μmol?L^–1)
Compd.	A-549	Hela	MCF-7	LO2
4a	27.54±0.04	>50	>50	>50
4b	10.55±0.09	18.93±0.03	11.97±0.04	45±0.13
4c	10.17±0.02	7.69±0.06	12.43±0.08	>50
4d	14.80±0.13	>50	>50	>50
4e	12.63±0.06	13.59±0.12	18.96±0.04	42.18±0.07
4f	7.50±0.06	49.99±0.03	18.90±0.04	>50
4g	16.77±0.08	38.13±0.12	17.71±0.12	>50
4h	17.68±0.12	6.83±0.06	7.87±0.03	33.12±0.20
4i	8.29±0.06	6.69±0.06	8.50±0.07	46.70±0.20
4j	10.32±0.09	>50	>50	>50
4k	12.23±0.10	27.07±0.06	42.19±0.10	>50
4l	14.41±0.02	49.99±0.02	>50	>50
4m	13.69±0.10	>50	>50	31.33±0.12
4n	9.25±0.07	15.64±0.09	21.30±0.14	30.25±0.22
4o	6.80±0.06	10.04±0.04	27.82±0.06	43.15±0.16
4p	13.48±0.03	30.96±0.06	29.25±0.05	>50
4q	9.00±0.11	5.32±0.12	6.73±0.08	>50
4r	6.75±0.05	7.93±0.03	4.51±0.03	45.13±0.20
Etoposide	9.86±0.05	8.68±0.06	6.44±0.10	16.74±0.03

Table 1. Antiproliferative activities of compounds 4a~4r

Figure 2. Cell cycle analysis of compound 4r in MCF-7 cells (a) Cell cycle distribution upon treatment with compound 4r at different concentrations (0, 2.5, 5, 10 μmol?L –1) for 24 h in MCF-7 cells; (b) The statistical analysis of MCF-7 cells in the different phases of cell cycle was presented

Figure 3. Cell apoptosis analysis of compound 4r in MCF-7 cells (a) Hoechst 33258 staining of MCF-7 cells treated with 0, 2.5, 5 and 10 μmol?L –1 of compound 4r for 24 h and fluorescence was observed by confocal microscopy; (b) cell apoptosis rate upon treatment with different concentrations of compound 4r for 24 h in MCF-7 cells; (c) graphical representation of effect of compound 4r on cell apoptosis

Figure 4. ROS production uantified by DCFH-DA fluorescence probe Image observations were carried out by fluorescence microscopy

Figure 5. ΔΨm quantified by using the JC-1 JC-1 staining of MCF-7 cells treated with 0, 2.5, 5 and 10 μmol?L –1 of compound 4r for 24 h and images were acquired by confocal microscopy

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