新型芳姜黄酮拼合吡咯螺环氧化吲哚类化合物的合成及其抗肿瘤活性

彭礼军, 周　根, 韩朔楠, 刘欢欢, 余章彪, 杨　超, 周　英, 赵　致, 刘雄利

(贵州大学 药学院 贵州省中药民族药创制工程中心，贵州 贵阳　550025)



·研究论文·

新型芳姜黄酮拼合吡咯螺环氧化吲哚类化合物的合成及其抗肿瘤活性

彭礼军, 周根, 韩朔楠, 刘欢欢, 余章彪*, 杨超, 周英, 赵致, 刘雄利*

(贵州大学 药学院 贵州省中药民族药创制工程中心，贵州 贵阳550025)

以取代靛红和肌氨酸为原料制得1,3-偶极子,再与(E)-芳姜烯酮类化合物在乙腈中经3+2环加成反应合成了10个新型的芳姜黄酮拼合吡咯螺环氧化吲哚类化合物(3a～3j)，产率70%～91%，d/r值15 ∶1～>20 ∶1,其结构经1H NMR,13C NMR和HR-MS(ESI-TOF)表征。采用MTT法研究了3a～3j对人肺癌细胞(A549)和人白血病细胞(K562)的体外抗肿瘤活性。结果表明：3f对K562抑制活性较好(IC50=31.1 μmol·L-1), 3b对A549抑制活性较好(IC50=54.1 μmol·L-1)。

取代靛红; 亚胺叶立德; 芳姜黄酮拼合吡咯螺环氧化吲哚; 1,3-偶极环加成反应; 合成; 抗肿瘤活性

3-吡咯螺环氧化吲哚具有广泛的生物活性[1-2]，如Spirotryprostatins B可完全抑制tsFT210细胞，阻断细胞分裂的G2/M期[3]。天然产物台钩藤碱具有调节大脑皮层M受体亚型和5-羟色胺受体的功能[4]。Alstonisine等也有类似功效[5]。

Scheme 1

芳姜黄酮类化合物，如倍半萜姜黄酮,(S)-芳姜黄酮，Bisacurone, Turmeronol B和 Turmeronol A等，均具有抗氧化、抗炎、抗癌、清除自由基、抗微生物、保护心脑血管系统及调节消化系统功能等多种药理作用[6-12]。

鉴于3-吡咯螺环氧化吲哚和芳姜黄酮类化合物良好的生物活性，根据药物设计拼合原理，将芳姜黄酮骨架拼合到3-吡咯螺环氧化吲哚骨架中，合成新型的芳姜黄酮拼合吡咯螺环氧化吲哚，可为生物活性筛选提供化合物源，对药物筛选有一定参考意义。本文以取代靛红(1a～1j)和肌氨酸为原料制得1,3-偶极子,再与(E)-芳姜烯酮类化合物(2a, 2f, 2g, 2i)在乙腈中经3+2环加成反应合成了10个新型的芳姜黄酮拼合吡咯螺环氧化吲哚类化合物(3a～3j, Scheme 1)，产率70%～91%，d/r值15 ∶1～>20 ∶1,其结构经1H NMR,13C NMR和HR-MS(ESI-TOF)表征。采用MTT法研究了3a～3j对人肺癌细胞(A549)和人白血病细胞(K562)的体外抗肿瘤活性。

1　实验部分

1.1仪器与试剂

WRS-1B型数字熔点仪；Bruker-400 MHz型核磁共振仪(CD3Cl为溶剂，TMS为内标); MicroTMQ-TOF型高分辨质谱仪。

所用试剂均为分析纯。

1.23a～3j的合成(以3a为例)

在反应管中依次加入N-甲基5-甲基靛红(1a)70.0 mg(0.4 mmol), (E)-芳姜烯酮(2a)120.0 mg(0.6 mmol)，肌氨酸71.2 mg(0.8 mmol)和乙腈15 mL，回流反应12 h。冷却至室温,减压蒸除溶剂,残余物经硅胶柱层析[洗脱剂：V(石油醚)∶V(乙酸乙酯)=4 ∶1]纯化得3a 136.7 mg。

用类似的方法合成3b～3j。

3a: 黄色固体,m.p.173.8～175.9 ℃;1H NMRδ: 1.39(s, 3H), 1.49(s, 3H), 2.03(s, 3H), 2.20(s, 6H), 2.23～2.29(m, 1H), 2.62～2.72(m, 1H), 3.13(s, 3H), 3.22～3.31(m, 1H), 3.35～3.47(m, 1H), 5.30(s, 1H), 6.74～6.79(m, 1H), 6.90(s, 2H), 7.00(s, 2H), 7.39(s, 2H);13C NMRδ: 20.3, 21.0, 26.2, 27.0, 35.1, 52.7, 56.4, 61.1, 68.1, 73.6, 107.3, 123.7, 126.8, 128.0, 129.2, 132.2, 136.0, 139.3, 141.3, 155.2, 177.7, 195.9; HR-MS(ESI-TOF)m/z: Calcd for C26H30N2O2Na{[M+Na]+}425.220 5, found 425.221 1。

3b: 淡黄色固体,m.p.185.1～187.2 ℃;1H NMRδ: 1.55(s, 3H), 1.71(s, 3H), 2.20(s, 3H), 2.31(s, 3H), 3.38(t,J=8.6 Hz, 1H), 3.53(t,J=8.8 Hz, 1H), 3.74(d,J=9.6 Hz, 1H), 4.25～4.32(m, 1H), 5.56(s, 1H), 6.82～6.85(m, 1H), 6.88～6.96(m, 2H), 7.13(d,J=7.8 Hz, 2H), 7.38(d,J=7.9 Hz, 2H), 9.17(s, 1H);13C NMRδ: 20.5, 21.0, 27.2, 35.1, 43.0, 60.9, 68.0, 74.3, 110.2, 114.1, 114.4, 115.3, 115.6, 123.5, 127.9, 129.2, 136.2, 136.7, 139.0, 156.7, 157.8, 160.2, 180.6, 195.4; HR-MS(ESI-TOF)m/z: Calcd for C24H25N2O2FNa{[M+Na]+}415.179 8, found 415.179 5。

3c: 白色固体,m.p.208.7～210.8 ℃;1H NMRδ: 1.50(s, 3H), 1.62(s, 3H), 2.20(s, 3H), 2.32(s, 3H), 2.34(s, 3H), 3.40(t,J=8.5 Hz, 1H), 3.54(t,J=8.8 Hz, 1H), 3.71(d,J=9.4 Hz, 1H), 4.28～4.35(m, 1H), 5.52(s, 1H), 6.93(t,J=7.5 Hz, 1H), 6.99～7.04 (m, 2H), 7.13(d,J=7.9 Hz, 2H), 7.41(d,J=8.0 Hz, 2H), 9.53(s, 1H);13C NMRδ: 16.2, 20.4, 21.0, 27.1, 35.2, 42.9, 53.4, 61.0, 68.1, 118.8, 122.7, 123.7, 123.8, 126.8, 128.0, 129.1, 130.3, 136.1, 139.5, 155.5, 181.1, 195.7; HR-MS(ESI-TOF)m/z: Calcd for C25H28N2O2Na{[M+Na]+} 411.204 8, found 411.204 8。

3d: 黄色固体,m.p.34.4～36.7 ℃;1H NMRδ: 1.53(s, 3H), 1.58(s, 3H), 2.11(s, 3H), 2.29(s, 3H), 2.53(s, 3H), 3.35(t,J=8.6 Hz, 1H), 3.51(d,J=7.2 Hz, 1H), 3.53(s, 3H), 3.66～3.69(m, 1H), 4.22～4.29(m, 1H), 5.41(s, 1H), 6.86～6.90(m, 2H), 6.98～7.01(m, 1H), 7.08～7.12(m, 2H), 7.36～7.40(m, 2H);13C NMRδ: 18.9, 20.2, 20.9, 27.0, 29.6, 35.0, 42.9, 61.0, 68.4, 119.0, 122.6, 123.6, 124.0, 127.9, 129.0, 132.6, 135.9, 139.1, 141.1, 155.0, 178.6, 195.7; HR-MS(ESI-TOF)m/z: Calcd for C26H30N2O2Na{[M+Na]+}425.220 5, found 425.220 5。

3e: 黄色固体,m.p.198.0～200.3 ℃;1H NMRδ: 1.56(s, 3H), 1.63(s, 3H), 2.21(s, 3H), 2.31(s, 3H), 3.38(t,J=8.5 Hz, 1H), 3.53(t,J=8.9 Hz, 1H), 3.72(d,J=9.6 Hz, 1H), 4.24～4.30(m, 1H), 5.54(s, 1H), 6.94～6.98(m, 1H), 7.07(d,J=7.3 Hz, 1H) 7.12(d,J=7.8 Hz, 2H), 7.18～7.20(m, 1H), 7.38(d,J=8.1 Hz, 2H), 8.54(s, 1H);13C NMRδ: 20.5, 21.0, 27.2, 35.2, 43.1, 60.9, 68.4, 74.9, 114.8, 123.5, 123.6, 124.8, 127.9, 129.2, 136.2, 138.3, 138.8, 156.5, 179.3, 195.4; HR-MS(ESI-TOF)m/z: Calcd for C24H25N2O2ClNa{[M+Na]+}431.150 2, found 431.150 0。

3f: 苍黄色固体,m.p.165.1～167.1 ℃;1H NMRδ: 1.39(s, 3H), 1.47(s, 3H), 2.01(s, 3H), 3.15(s, 3H), 3.25～3.34(m, 1H), 3.45(t,J=8.6 Hz, 1H), 3.58～3.69(m, 1H), 4.21～4.25(m, 1H), 5.32(s, 1H), 6.67(s, 1H), 6.89～6.94(m, 2H), 7.09(d,J=7.5 Hz, 2H), 7.08～7.19(m, 2H), 7.40(s, 2H);13C NMRδ: 20.3, 26.2, 27.1, 35.1, 43.3, 56.9, 61.1, 68.2, 73.8, 107.6, 122.8, 123.6, 126.7, 128.1, 128.5, 129.1, 135.6, 142.5, 143.6, 155.6, 177.6, 195.7; HR-MS(ESI-TOF)m/z: Calcd for C24H26N2O2Na{[M+Na]+}397.189 2, found 397.189 6。

3g: 黄色固体,m.p.177.2～179.4 ℃;1H NMRδ: 1.55(s, 3H), 1.76(s, 3H), 2.21(s, 3H), 3.35～3.40(m, 1H), 3.46～3.52(m, 1H), 3.60～3.65(m, 1H), 4.24～4.31(m, 1H), 5.51(s, 1H), 6.74～6.78(m, 1H), 7.24～7.29(m, 3H), 7.31～7.35(m, 1H), 7.39～7.44(m, 2H), 8.82(brs, 1H);13C NMRδ: 20.7, 27.3, 35.0, 42.9, 60.6, 68.0, 73.8, 111.0, 115.7, 123.2, 128.7, 129.4, 129.5, 132.0, 132.5, 139.8, 140.4, 157.5, 180.0, 195.1; HR-MS(ESI-TOF)m/z: Calcd for C23H22N2O2ClBrNa{[M+Na]+}495.045 1, found 495.045 1。

3h: 黄色固体,m.p.46.5～49.0 ℃;1H NMRδ: 1.52(s, 3H), 1.73(s, 3H), 2.13(s, 3H), 3.24(s, 3H), 3.33～3.38(m, 1H), 3.47～3.52(m, 1H), 3.57～3.61(m, 1H), 4.22～4.29(m, 1H), 5.37(s, 1H), 6.64～6.68(m, 1H), 7.24～7.28(m, 3H), 7.35～7.39(m, 1H), 7.40～7.44(m, 2H);13C NMRδ: 20.6, 26.2, 27.2, 34.9, 42.9, 60.6, 68.0, 73.3, 108.9, 115.7, 123.3, 128.6, 129.1, 129.5, 131.9, 132.3, 140.3, 142.7, 156.9, 177.3, 195.1; HR-MS(ESI-TOF)m/z: Calcd for C24H24N2O2ClBrNa{[M+Na]+}509.060 7, found 509.060 7。

3i: 黄色固体,m.p.55.1～57.3 ℃;1H NMRδ: 1.88(s, 3H), 2.16(s, 6H), 2.81(s, 3H), 3.10(s, 1H), 3.17～3.26(m, 1H), 3.44～3.49(m, 1H), 3.59(s, 6H), 3.72(s, 3H), 4.12～4.18(m, 1H), 6.11(s, 1H), 6.54～6.58(m, 1H), 7.10～7.15(m, 1H), 7.21～7.31(m, 3H), 7.46～7.51(m, 1H);13C NMRδ: 21.1, 25.1, 27.9, 31.1, 35.8, 52.1, 55.8, 56.1, 56.8, 60.7, 103.2, 104.9, 107.7, 122.7, 123.2, 127.9, 129.0, 131.1, 136.8, 144.4, 152.3, 153.3, 157.4, 176.2, 199.5; HR-MS(ESI-TOF)m/z: Calcd for C27H32N2O5Na{[M+Na]+}487.220 9, found 487.220 9。

3j: 黄色固体,m.p.66.3～68.5 ℃;1H NMRδ: 1.50(s, 3H), 1.59(s, 3H), 2.18(s, 3H), 3.34(t,J=8.5 Hz, 1H), 3.49(t,J=8.8 Hz, 1H), 3.67(d,J=5.6 Hz, 1H), 3.73(s, 3H), 3.79(s, 6H), 4.19～4.26(m, 1H), 5.54(s, 1H), 6.68(s, 2H), 6.98(d,J=7.5 Hz, 1H), 7.09(t,J=7.6 Hz, 2H), 7.16(d,J=7.3 Hz, 1H), 7.27～7.37(m, 3H), 7.47(t,J=7.6 Hz , 2H);13C NMRδ: 20.5, 27.4, 35.1, 43.9, 56.2, 60.8, 61.0, 68.5, 73.6, 103.4, 104.9, 105.2, 109.0, 123.3, 123.9, 126.3, 126.5, 126.6, 128.3, 129.0, 129.4, 129.7, 134.3, 136.6, 138.1, 143.5, 153.2, 156.4, 177.3, 195.9; HR-MS(ESI-TOF)m/z: Calcd for C32H34N2O5Na{[M+Na]+}549.236 5, found 549.236 5。

1.3体外抗肿瘤活性测试

采用MTT法[11-12]测试了3a～3j对人肺癌细胞(A549)和人白血病细胞(K562)的体外抗肿瘤活性,以顺铂为阳性对照药。

2　结果与讨论

2.1合成

通过底物扩展，我们发现该反应的活性普遍较高，12 h内基本反应完全(TLC检测)。其中，芳姜烯酮苯环上为吸电子氯原子取代产物，产率最高(3h, 91%)；芳姜烯酮苯环上为大位阻3,4,5-三甲氧基取代产物，产率明显降低(3j, 70%)。

2.2抗肿瘤活性

表1为3a～3j对K562和A549的体外抗肿瘤活性。由表1可见，3a～3j对A549和K562的抑制活性均弱于顺铂，但可作为先导化合物的骨架进一步研究。

表1　3a～3j的体外抗肿瘤活性

合成了10个新型的芳姜黄酮拼合吡咯螺环氧化吲哚类化合物(3a～3j)。并用MTT法研究了3a～3j对人肺癌细胞(A549)和人白血病细胞(K562)的体外抗肿瘤活性。结果表明：3f对K562抑制活性较好(IC50=31.1 μmol·L-1), 3b对A549抑制活性较好(IC50=54.1 μmol·L-1)。虽然3的活性弱于顺铂，但可作为先导化合物骨架进一步研究。其他相关药理活性的研究正在进行中。

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Synthesis and Antitumor Activities of Novel Turmerone Motif-fused Spiropyrrolidine Oxindoles

PENG Li-jun,ZHOU Gen,HAN Shuo-nan,LIU Huan-huan,YU Zhang-biao*,YANG Chao,ZHOU Ying,ZHAO Zhi,LIU Xiong-li*

(Guizhou Engineering Center for Innovative Traditional Chinese Medicine and Ethnic Medicine,College of Pharmacy, Guizhou University, Guiyang 550025, China)

Ten novel turmerone motif-fused spiropyrrolidine oxindoles(3a～3j) were synthesized by 1,3-dipolar reaction of substituted isatins with sarcosine, then 3+2 cycloaddition with (E)-dienones in MeCN. The yields andd/rof 3a～3j were 70%～91% and 15 ∶1～>20 ∶1, respectively. The structures were characterized by1H NMR,13C NMR and HR-MS(ESI-TOF). Theinvitroantitumor activities against human lung cancer cells(A549) and human leukemia cells(K562) were demonstrated by MTT assays. The results showed that 3f exhibited best activity against K562 with IC50of 31.1 μmol·L-1and 3b indicated best activity against A549 with IC50of 54.1 μmol·L-1.

substituted isatin; azomethine ylide; turmerone motif-fused spiropyrrolidine oxindole; 1,3-dipolar cycloaddition reaction; synthesis; antitumor activity

2016-04-30

国家自然科学基金青年基金资助项目(21302024); 国家自然科学基金地区基金资助项目(81560563); 贵州省教学改革创新项目(SJJG201423); 贵州省制药工程专业学位研究生工作站(黔教研合JYSZ字【2014】002)

彭礼军(1987-)，男，汉族，湖南常德人，硕士研究生，主要从事天然产物活性成分研究。 E-mail: lijunpe@163.com

通信联系人： 刘雄利，博士，副教授, E-mail: ls.liuxl@gzu.edu.cn; 余章彪，教授, E-mail: gym.zbyu@gzu.edu.cn

O626.13; O623.7

A

10.15952/j.cnki.cjsc.1005-1511.2016.08.16117