彭禮軍, 周　根, 韓朔楠, 劉歡歡, 余章彪, 楊　超, 周　英, 趙　致, 劉雄利

(貴州大學(xué) 藥學(xué)院 貴州省中藥民族藥創(chuàng)制工程中心，貴州 貴陽(yáng)　550025)

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·研究論文·

新型芳姜黃酮拼合吡咯螺環(huán)氧化吲哚類(lèi)化合物的合成及其抗腫瘤活性

彭禮軍, 周根, 韓朔楠, 劉歡歡, 余章彪*, 楊超, 周英, 趙致, 劉雄利*

(貴州大學(xué) 藥學(xué)院 貴州省中藥民族藥創(chuàng)制工程中心，貴州 貴陽(yáng)550025)

以取代靛紅和肌氨酸為原料制得1,3-偶極子,再與(E)-芳姜烯酮類(lèi)化合物在乙腈中經(jīng)3+2環(huán)加成反應(yīng)合成了10個(gè)新型的芳姜黃酮拼合吡咯螺環(huán)氧化吲哚類(lèi)化合物(3a～3j)，產(chǎn)率70%～91%，d/r值15 ∶1～>20 ∶1,其結(jié)構(gòu)經(jīng)1H NMR,13C NMR和HR-MS(ESI-TOF)表征。采用MTT法研究了3a～3j對(duì)人肺癌細(xì)胞(A549)和人白血病細(xì)胞(K562)的體外抗腫瘤活性。結(jié)果表明：3f對(duì)K562抑制活性較好(IC50=31.1 μmol·L-1), 3b對(duì)A549抑制活性較好(IC50=54.1 μmol·L-1)。

取代靛紅; 亞胺葉立德; 芳姜黃酮拼合吡咯螺環(huán)氧化吲哚; 1,3-偶極環(huán)加成反應(yīng); 合成; 抗腫瘤活性

3-吡咯螺環(huán)氧化吲哚具有廣泛的生物活性[1-2]，如Spirotryprostatins B可完全抑制tsFT210細(xì)胞，阻斷細(xì)胞分裂的G2/M期[3]。天然產(chǎn)物臺(tái)鉤藤堿具有調(diào)節(jié)大腦皮層M受體亞型和5-羥色胺受體的功能[4]。Alstonisine等也有類(lèi)似功效[5]。

Scheme 1

芳姜黃酮類(lèi)化合物，如倍半萜姜黃酮,(S)-芳姜黃酮，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、保護(hù)心腦血管系統(tǒng)及調(diào)節(jié)消化系統(tǒng)功能等多種藥理作用[6-12]。

鑒于3-吡咯螺環(huán)氧化吲哚和芳姜黃酮類(lèi)化合物良好的生物活性，根據(jù)藥物設(shè)計(jì)拼合原理，將芳姜黃酮骨架拼合到3-吡咯螺環(huán)氧化吲哚骨架中，合成新型的芳姜黃酮拼合吡咯螺環(huán)氧化吲哚，可為生物活性篩選提供化合物源，對(duì)藥物篩選有一定參考意義。本文以取代靛紅(1a～1j)和肌氨酸為原料制得1,3-偶極子,再與(E)-芳姜烯酮類(lèi)化合物(2a, 2f, 2g, 2i)在乙腈中經(jīng)3+2環(huán)加成反應(yīng)合成了10個(gè)新型的芳姜黃酮拼合吡咯螺環(huán)氧化吲哚類(lèi)化合物(3a～3j, Scheme 1)，產(chǎn)率70%～91%，d/r值15 ∶1～>20 ∶1,其結(jié)構(gòu)經(jīng)1H NMR,13C NMR和HR-MS(ESI-TOF)表征。采用MTT法研究了3a～3j對(duì)人肺癌細(xì)胞(A549)和人白血病細(xì)胞(K562)的體外抗腫瘤活性。

1　實(shí)驗(yàn)部分

1.1儀器與試劑

WRS-1B型數(shù)字熔點(diǎn)儀；Bruker-400 MHz型核磁共振儀(CD3Cl為溶劑，TMS為內(nèi)標(biāo)); MicroTMQ-TOF型高分辨質(zhì)譜儀。

所用試劑均為分析純。

1.23a～3j的合成(以3a為例)

在反應(yīng)管中依次加入N-甲基5-甲基靛紅(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，回流反應(yīng)12 h。冷卻至室溫,減壓蒸除溶劑,殘余物經(jīng)硅膠柱層析[洗脫劑：V(石油醚)∶V(乙酸乙酯)=4 ∶1]純化得3a 136.7 mg。

用類(lèi)似的方法合成3b～3j。

3a: 黃色固體,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: 淡黃色固體,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: 白色固體,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: 黃色固體,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: 黃色固體,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: 蒼黃色固體,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: 黃色固體,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: 黃色固體,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: 黃色固體,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: 黃色固體,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體外抗腫瘤活性測(cè)試

采用MTT法[11-12]測(cè)試了3a～3j對(duì)人肺癌細(xì)胞(A549)和人白血病細(xì)胞(K562)的體外抗腫瘤活性,以順鉑為陽(yáng)性對(duì)照藥。

2　結(jié)果與討論

2.1合成

通過(guò)底物擴(kuò)展，我們發(fā)現(xiàn)該反應(yīng)的活性普遍較高，12 h內(nèi)基本反應(yīng)完全(TLC檢測(cè))。其中，芳姜烯酮苯環(huán)上為吸電子氯原子取代產(chǎn)物，產(chǎn)率最高(3h, 91%)；芳姜烯酮苯環(huán)上為大位阻3,4,5-三甲氧基取代產(chǎn)物，產(chǎn)率明顯降低(3j, 70%)。

2.2抗腫瘤活性

表1為3a～3j對(duì)K562和A549的體外抗腫瘤活性。由表1可見(jiàn)，3a～3j對(duì)A549和K562的抑制活性均弱于順鉑，但可作為先導(dǎo)化合物的骨架進(jìn)一步研究。

表1　3a～3j的體外抗腫瘤活性

合成了10個(gè)新型的芳姜黃酮拼合吡咯螺環(huán)氧化吲哚類(lèi)化合物(3a～3j)。并用MTT法研究了3a～3j對(duì)人肺癌細(xì)胞(A549)和人白血病細(xì)胞(K562)的體外抗腫瘤活性。結(jié)果表明：3f對(duì)K562抑制活性較好(IC50=31.1 μmol·L-1), 3b對(duì)A549抑制活性較好(IC50=54.1 μmol·L-1)。雖然3的活性弱于順鉑，但可作為先導(dǎo)化合物骨架進(jìn)一步研究。其他相關(guān)藥理活性的研究正在進(jìn)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

國(guó)家自然科學(xué)基金青年基金資助項(xiàng)目(21302024); 國(guó)家自然科學(xué)基金地區(qū)基金資助項(xiàng)目(81560563); 貴州省教學(xué)改革創(chuàng)新項(xiàng)目(SJJG201423); 貴州省制藥工程專(zhuān)業(yè)學(xué)位研究生工作站(黔教研合JYSZ字【2014】002)

彭禮軍(1987-)，男，漢族，湖南常德人，碩士研究生，主要從事天然產(chǎn)物活性成分研究。 E-mail: lijunpe@163.com

通信聯(lián)系人： 劉雄利，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