有机化学  2016, Vol. 36 Issue (3): 630-637   PDF    
去氢表雄酮噻唑衍生物的合成及抗肿瘤活性评估
崔建国a,b , 赵丹丹a , 何冬梅a , 黄燕敏a , 刘志平a , 林啟福b , 石海信b , 甘春芳a     
a 广西师范学院化学与材料科学学院 北部湾环境演变与资源利用教育部重点实验室 南宁 530001;
b 广西钦州学院 广西高校北部湾石油天然气资源有效利用重点实验室 钦州 535000
国家自然科学基金(Nos. 21462009, 21562007)、广西高校北部湾石油天然气资源有效利用重点实验室开放课题(No. 2014KLOG09)、合成与天然功能分子化学广西重点实验室资助项目
*通讯作者：甘春芳, E-mail: ganchunfang2008@126.com
摘要：以去氢表雄酮为原料, 通过微波一步合成法及常规2步合成方法合成得到16个新的具有不同结构特征的去氢表雄酮噻唑衍生物.所有合成物都通过了IR、NMR及HRMS的结构表征.另外, 分别选用人宫颈癌细胞(HeLa)、人肝癌细胞(HepG)、人肺癌细胞(A549)和人正常肾上皮细胞(HEK-293T)对合成物的抗肿瘤活性进行了研究, 结果表明4-(4'-三氟甲基)苯基噻唑-2-去氢表雄酮腙(4)对所测试的肿瘤细胞株具有很好的生长增殖抑制活性, 对上述肿瘤细胞的IC50值分别为13.2、11.3和8.3 μmol·L-1, 对人肾上皮正常细胞HEK-293T却没有表现出明显的抑制作用(IC50值>100 μmol·L-1).但是, 其它合成物对所测试的细胞株没有表现出明显的抑制活性.
关键词：去氢表雄酮    噻唑    去氢表雄酮噻唑衍生物    抗肿瘤活性    
Synthesis of Dehydroepiandrosteronyl Thiazole Derivatives and Their Antiproliferative Evaluation
Cui Jianguoa,b , Zhao Dandana , He Dongmeia , Huang Yanmina , Liu Zhipinga , Lin Qifub , Shi Haixinb , Gan Chunfanga     
a Key Laboratory of Environment Change and Resources Use in Beibu Gulf Ministry of Education, Chemistry and Materials Science College, Guangxi Teachers Education University, Nanning 530001 ;
b Guangxi Colleges and University Key Laboratory of Beibu Gulf Oil and Natural Gas Resoure Effective Utilization, Qizhou University, Qinzhou 535000
Received: September 18, 2015; published: November 18, 2015; published online: November 26, 2015
Project supported by the National Natural Science Foundation of China (Nos. 21462009, 21562007), the Key Laboratory Project of Effective Utilization of Petroleum and Natural Gas Resources in Guangxi Beibu Gulf (No. 2014KLOG09), and the Synthetic and Natural Functional Molecular Chemistry Lab of Guangxi
Abstract: Using dehydroepiandrosterone as starting material, 16 dehydroepiandrosteronyl thiazol derivatives with different structures had been synthesized via microwave irradiation assisted one-pot reaction and general synthetic method, and their structures were characterized by IR, NMR and HRMS. The antiproliferative activity of the compounds was evaluated against human cervical carcinoma (HeLa), human lung carcinoma (A549), human liver carcinoma cells (HEPG2) and normal kidney epithelial cells (HEK293T). The results showed that compound 4 displayed significant inhibitory activity to tested cancer cells, and the values of IC50 were 13.2, 11.3 and 8.3 μmol·L-1 respectively, but was almost inactive to HEK293T cells (IC50>100 μmol·L-1). However, other compounds did not exhibit distinct antiproliferative activity to all tested cells.
Key Words: dehydroepiandrosterone    thiazole    dehydroepiandrosteronyl thiazole derivatives    antiproliferative activity    

甾体化合物由于其特殊的化学结构而表现出许多不同的生物活性, 它在生命体中起着十分重要的作用.除了作为生命体中的荷尔蒙在生命体中发挥重要作用外, 以甾体为母体的甾体药物目前在临床上已经得到广泛应用, 如抗炎、抗菌、抗肿瘤、降压、降胆固醇及利尿类药物等[1～8].以甾体结构为母体, 外接一个芳杂环结构后所得到的化合物会出现与甾体母体本身不同的生理活性, 它们常常会表现出不同的抗肿瘤、抗菌或抗病毒活性[9].

噻唑环是一类重要的含N、S杂原子的五元芳杂环, 其特殊的结构特征使得噻唑类化合物具有广泛的生物功能, 它是一类具有广谱生物活性的杂环化合物, 目前在化学、药学、生物学和材料科学等诸多领域已获得广泛应用[10].对噻唑衍生物的研究开发已成为当今医药、农药研究的热点之一, 在医药应用上噻唑类化合物对多种疾病具有独特的疗效, 如在抗菌、抗真菌、抗结核、抗癌、抗病毒、降血糖、消炎镇痛、抗癫痫、抗寄生虫等药物中的应用[11～15]; 在农药应用上, 噻唑类化合物具有杀虫、杀菌和除草活性, 并表现出高效、低毒的特点[16, 17].因此, 噻唑类化合物具有广阔的研究开发前景.

Shingate等曾在甾核支链的20-位上引入噻唑环, 得到一系列甾体噻唑衍生物, 其中某些化合物对白色念珠菌显示出良好的抑制活性, 其最低抑制浓度(MIC)值优于临床一线药物氟康唑[18].在前期研究中, 我们曾在一些具有不同甾核结构的甾体化合物中引入不同的杂环药效团, 并研究了合成物的体外抑制肿瘤细胞生长增殖活性, 结果表明其中一些化合物具有很好的抑制肿瘤细胞增殖功效[19～22].

去氢表雄酮(Dehydroepiandrosterone, DHEA)是20世纪的重要发现之一, 它是人体生物合成多种性激素中的前体物质.去氢表雄酮能抑制过多线粒体和5-磷酸核糖酯, 从而抑制和预防肿瘤生长. DHEA具有明显的抗促癌活性, 它可明显抑制促癌剂巴豆油引起的小鼠急性耳肿胀, 对巴豆油诱发的表皮细胞(ODC)活性升高具有显著抑制作用[23].在本研究中, 我们以DHEA作为母体, 在其甾核的17-位支链中引入噻唑环药效团, 从而合成得到系列去氢表雄酮噻唑衍生物, 进一步研究合成物的抗肿瘤活性, 试图探索发现一些具有更好抗肿瘤活性的甾体化合物, 为新型抗肿瘤药物的研究开发提供理论参考.

2 结果与讨论
2.1 化学部分

微波合成是一种绿色环保、反应时间短、后处理简单的有机合成方法, 近年来已经在有机合成领域得到广泛应用. Ahluwalia等[24]曾报道了一种通过微波辐射制备噻唑芳腙类化合物的合成方法; 张英群等[25]通过含不同取代基的芳香醛与氨基硫脲缩合生成缩氨基硫脲, 然后进一步与α-溴代乙酰基香豆素经微波辐射合成某些具有抗菌活性的香豆素噻唑芳醛腙化合物.在这里, 我们直接通过去氢表雄酮、不同的4-取代-α-溴代苯乙酮及氨基硫脲在微波辐射下进行一锅化反应, 成功合成得到4-取代苯基噻唑去氢表雄酮腙(2～7) (Eq. 1).

(1)

从化合物2的红外光谱中可以看到, 在2222 cm－1波数处对应的吸收峰是CN的伸缩振动吸收峰.在1H NMR谱中, δ 7.00处的单峰是噻唑环上5-位氢的化学位移, δ 7.67和7.88的双重峰分别对应于苯环上3', 5'-位和2', 6'-位两组等位氢的化学位移, 结合在13C NMR中δ 106.5～169.9之间出现的噻唑环上碳、苯环上碳及氰基碳的吸收峰, 可说明苯代噻唑结构已经与去氢表雄酮相连接; 而δ 165.8可认定为C＝N双键的化学位移, 由此判断去氢表雄酮的17-位羰基已经转换成为腙基.进一步结合其HRMS, 可以确定所得到的化合物2结构为4-(4'-氰基苯基)噻唑-2-去氢表雄酮腙.其它化合物3～7的结构类似地得到表征.

以化合物4的合成为对象, 我们对微波反应的实验条件进行了探索, 分别对反应的溶剂量、微波辐射功率、反应时间及反应温度进行了优化.最后, 确定反应溶剂乙醇15 mL、微波功率300 W、反应温度85 ℃及反应时间30 min作为此反应的最佳反应条件.

另外, 我们采用4-甲基氨基硫脲及4-苯基氨基硫脲代替氨基硫脲, 试图利用Eq. 1的微波促进一锅法的合成方法制备类似的去氢表雄酮噻唑衍生物, 但是没有获得成功.原因可能是由于氨基硫脲中的4-氨基发生取代后, 增加了反应中分子之间接触的空间位阻, 使反应发生的难度增加.但是, 使用Scheme 1的分步合成方法, 我们成功地得到了目标产物9～17.

图式1 化合物9～17的合成路线 Scheme1 The synthetic route of compounds9～17

首先, 去氢表雄酮与4-甲基氨基硫脲或4-苯基氨基硫脲反应, 分别得到相应的N-甲基去氢表雄酮-17-缩氨硫腙和N-苯基去氢表雄酮-17-缩氨硫腙, 然后N-甲基去氢表雄酮-17-缩氨硫腙分别和不同的苯基取代-α-溴代苯乙酮反应, 得到去氢表雄酮N-甲基-4-芳基噻唑-2(3H)-吖嗪化合物9～12.同样地, N-苯基去氢表雄酮-17-缩氨硫腙和不同的苯基取代-α-溴代苯乙酮反应得到相应的去氢表雄酮噻唑衍生物13～17.所有合成物的结构都通过了IR、NMR及HRMS的结构表征.

另外, 化合物3中的硝基进一步用二氯化锡还原成为氨基(Eq. 2), 得到化合物18, 18的结构同样得到了相应光谱及波谱的结构表征.

(2)
2.2 生物部分

为了探索所合成化合物的抗肿瘤活性, 分别选用人宫颈癌细胞(HeLa)、人肝癌细胞(HepG)、人肺癌细胞(A549)和人肾上皮细胞(HEK-293T)作为研究对象, 使用所合成的目标化合物对上述肿瘤细胞株进行了体外抑制肿瘤细胞生长增值活性研究.结果如表 1所示.

表 1 去氢表雄酮噻唑化合物的体外抗增殖活性[IC50/(μmol• L－1)]a Table 1 The antiproliferative activities of dehydroepiandrosteronyl thiazole derivatives [IC50/(μmol•L－1)]

从表 1中数据可以看到, 除了化合物4外, 此类去氢表雄酮噻唑衍生物对所测试的肿瘤细胞株和人体正常细胞293T都没有表现出明显的抑制活性.然而, 有趣的是对于化合物4来说, 其对所测试的肿瘤细胞株都显示出很好的抑制活性, 如对肺癌细胞A549的抑制活性IC50值仅为8.3 μmol•L－1, 但是对人体正常肾上皮细胞(HEK-293T)却基本上没有显示出抑制活性(IC50值＞100 μmol•L－1); 其对HeLa、HepG和A549细胞的SI值(化合物对正常细胞与肿瘤细胞抑制的比值)分别大于7.6、8.8和12, 说明该化合物对正常细胞具有较小的伤害, 值得进一步的深入研究.

3 结论

以去氢表雄酮作为原料, 通过微波一步合成法及常规2步合成方法合成得到16个具有不同结构特征的去氢表雄酮噻唑衍生物, 并分别选用某些肿瘤细胞株和人正常肾上皮细胞(HEK-293T)对合成物的抗肿瘤活性进行了研究, 结果表明4-(4'-三氟甲基)苯基噻唑-2-去氢表雄酮腙(4)对所测试的肿瘤细胞株具有很好的生长增殖抑制活性, 却对人肾上皮正常细胞HEK-293T没有表现出明显的抑制作用.但是, 其它合成物则对所测试的细胞株没有表现出明显的抑制活性.这些研究结果为新的甾体抗肿瘤药物的设计合成提供了有用的参考.

4 实验部分
4.1 仪器与试剂

熔点测定采用X6显微熔点测定仪, 温度计未经校正; IR光谱采用Thermo Scientific Nicolet IS-10傅立叶红外光谱仪测定; 1H NMR和13C NMR谱采用德国Bruker AV-300型或Bruker AV-600型超导核磁共振仪测定; 高分辨质谱采用美国Agilent 6210 TOFMS质谱仪测定; 细胞增殖活性评估采用MTT方法, 使用MLLTISKAN MK3分光仪测定.去氢表雄酮及其它试剂均购于百灵威科技有限公司; 所用试剂如无特别说明均为分析纯, 溶剂按常规方法进行处理干燥.

4.2 实验方法
4.2.1 化合物2～7的一般合成步骤

分别称取去氢表雄酮0.40 mmol、氨基硫脲0.42 mmol和4-取代-α-溴代苯乙酮0.43 mmol置于100 mL双颈瓶中.加入15 mL无水乙醇使其完全溶解后置于微波合成反应器中, 装好回流冷凝管, 插入温度传感器, 功率调至300 W, 预设温度为85 ℃, 在此条件下反应30 min. TLC跟踪检测无原料点后停止反应, 冷却至室温后有浅黄色粉末状固体析出, 减压蒸馏出大部分溶剂, 加入30 mL蒸馏水, 用乙酸乙酯萃取3次, 合并有机相.有机相分别用饱和碳酸氢钠、饱和氯化钠溶液洗涤, 无水硫酸钠干燥.减压蒸出溶剂后, 得到的固体通过柱层析分离[洗脱剂: V(二氯甲烷):V(乙酸乙酯)＝50:1]得到目标产物.

4-(4'-氰基)苯基噻唑-2-去氢表雄酮腙(2):浅黄色固体, 产率53%. m.p. 284～285 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.96 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 3.61～3.51 (m, 1H, C3-αH), 5.39 (d, J＝4.5 Hz, 1H, C6-H), 7.00 (s, 1H, 5-thiazole-H), 7.67 (d, J＝8.4 Hz, 2H, 3', 5'-PhH), 7.88 (d, J＝8.4 Hz, 2H, 2', 6'-PhH), 8.21 (br s, 1H, NH); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.4 (19-C), 20.6 (11-C), 23.6 (15-C), 25.9 (16-C), 31.3 (8-C), 31.4 (2-C), 31.6 (7-C), 34.0 (12-C), 36.7 (1-C), 37.2 (10-C), 42.2 (4-C), 44.5 (13-C), 50.4 (9-C), 53.9 (14-C), 71.6 (3-C), 106.5 (5-thiazole-C), 119.0 (CN), 121.0 (6-C), 110.7, 126.3, 132.5, 139.0 (Ph-C), 141.1 (5-C), 149.3 (4-thiazole-C), 165.8 (17-C), 169.9 (2-thiazole-C); IR (KBr) ν: 3473, 2942, 2222, 1604, 1556, 1439, 1399, 1297, 1100, 1033, 843, 741 cm－1; HREIMS calcd for C29H34N4-OS [M＋H]＋ 487.2536, found 487.2532.

4-(4'-硝基)苯基噻唑-2-去氢表雄酮腙(3):黄色固体, 产率63%. m.p. 286～287 ℃; 1H NMR (300 MHz, DMSO-d6) δ: 0.85 (s, 3H, 18-CH3), 0.97 (s, 3H, 19-CH3), 3.30～3.21 (m, 1H, C3-αH), 4.66 (s, 1H, OH), 5.28 (d, J＝3.6 Hz, 1H, C6-H), 7.58 (s, 1H, 5-thiazole-H), 8.08 (d, J＝9.0 Hz, 2H, 3', 5'-PhH), 8.24 (d, J＝9.0 Hz, 2H, 2', 6'-PhH), 10.68 (br s, 1H, NH); 13C NMR (75 MHz, DMSO) δ: 17.3 (18-C), 19.6 (19-C), 20.6 (11-C), 23.6 (15-C), 27.4 (16-C), 31.2 (8-C), 31.4 (2-C), 31.8 (7-C), 34.3 (12-C), 36.7 (1-C), 37.4 (10-C), 42.6 (4-C), 44.4 (13-C), 50.4 (9-C), 53.7 (14-C), 70.5 (3-C), 108.5 (5-thiazole-C), 120.5 (6-C), 124.5, 126.7, 141.4, 146.5 (Ph-C), 141.9 (5-C), 148.9 (4-thiazole-C), 166.8 (17-C), 170.9 (2-thiazole-C); IR (KBr) ν: 3471, 2925, 1594, 1556, 1442, 1339, 1262, 1105, 1038, 851, 778, 711 cm－1; HREI-MS calcd for C28H34N4O3S [M＋H]＋ 507.2427, found 507.2430.

4-(4'-三氟甲基)苯基噻唑-2-去氢表雄酮腙(4):浅黄色固体, 产率54%. m.p. 231～233 ℃; 1H NMR (600 MHz, CDCl3) δ: 0.94 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 2.41 (dd, J＝18.0, 9.0 Hz, 1H, C4-βH), 3.57～3.51 (m, 1H, C3-αH), 5.37 (d, J＝4.8 Hz, 1H, C6-H), 6.94 (s, 1H, 5-thiazole-H), 7.62 (d, J＝8.4, 2H, 3', 5'-PhH), 7.87 (d, J＝8.4 Hz, 2H, 2', 6'-PhH), 8.16 (br s, 1H, NH); 13C NMR (150 MHz, CDCl3) δ: 17.0 (18-C), 19.5 (19-C), 20.6 (11-C), 23.7 (16-C), 26.0 (15-C), 31.4 (2-C), 31.5 (7-C), 31.7 (8-C), 34.1 (12-C), 36.7 (10-C), 37.3 (1-C), 42.3 (4-C), 44.5 (13-C), 50.5 (9-C), 54.0 (14-C), 71.8 (3-C), 105.5 (5-thiazole-C), 121.2 (6-C), 125.7 (CF3), 125.7, 125.8, 126.1, 138.2 (Ph-C), 141.1 (5-C), 149.8 (4-thiazole-C), 165.7 (17-C), 169.9 (2-thiazole-C); IR (KBr) ν: 3431, 2940, 1728, 1656, 1556, 1374, 1322, 1247, 1125, 1068, 705 cm－1; HREIMS calcd for C29H34F3N3OS [M＋H]＋ 530.2455, found 530.2447.

4-(4'-甲氧基)苯基噻唑-2-去氢表雄酮腙(5):浅黄色固体, 产率74%. m.p. 235～237 ℃; 1H NMR (600 MHz, CDCl3) δ: 0.93 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 3.57～3.51 (m, 1H, C3-αH), 3.82 (s, 3H, OCH3), 6.65 (s, 1H, C6-H), 6.90 (d, J＝9.0 Hz, 2H, 3', 5'-PhH), 6.99 (s, 1H, 5-thiazole-H), 7.69 (d, J＝9.0 Hz, 2H, 2', 6'-PhH), 8.50 (br s, 1H, NH); 13C NMR (150 MHz, CDCl3) δ: 17.0 (18-C), 19.6 (19-C), 20.7 (11-C), 23.8 (15-C), 29.2 (16-C), 31.4 (8-C), 31.5 (2-C), 31.7 (7-C), 34.1 (12-C), 36.8 (1-C), 37.3 (10-C), 42.4 (4-C), 44.6 (13-C), 50.5 (9-C), 54.0 (14-C), 55.5 (OCH3) 71.8 (3-C), 101.3 (5-thiazole-C), 121.2 (6-C), 114.1, 122.0, 127.3, 164.4 (Ph-C), 141.2 (5-C), 159.5 (4-thiazole-C), 166.4 (17-C), 169.7 (2-thiazole-C); IR (KBr) ν: 3446, 2927, 1733, 1651, 1561, 1484, 1337, 1240, 1167, 1029, 704 cm－1; HREIMS calcd for C29H37N3O2S [M＋H]＋ 492.2679, found 492.2679.

4-(4'-氯)苯基噻唑-2-去氢表雄酮腙(6):浅黄色固体, 产率38%. m.p. 269～271 ℃; 1H NMR (300 MHz, DMSO) δ: 0.85 (s, 3H, 18-CH3), 0.98 (s, 3H, 19-CH3), 3.45～3.35 (m, 1H, C3-αH), 4.70 (s, 1H, OH), 5.29 (d, J＝4.8 Hz, 1H, C6-H), 7.24 (s, 1H, 5-thiazole-H), 7.43 (d, J＝8.4 Hz, 2H, 3', 5'-PhH), 7.84 (d, J＝8.4 Hz, 2H, 2', 6'-PhH), 10.51 (br s, 1H, -NH); 13C NMR (75 MHz, CDCl3) δ: 13.6 (18-C), 19.4 (19-C), 20.4 (11-C), 23.6 (15-C), 29.7 (16-C), 31.4 (8-C), 31.5 (2-C), 31.6 (7-C), 34.0 (12-C), 36.7 (1-C), 37.2 (10-C), 42.2 (4-C), 44.4 (13-C), 50.2 (9-C), 53.9 (14-C), 71.6 (3-C), 103.6 (5-thiazole-C), 120.9 (6-C), 121.0, 127.1, 128.7, 133.3 (Ph-C), 141.0 (5-C), 149.6 (4-thiazole-C), 165.7 (17-C), 169.7 (2-thiazole-C); IR (KBr) ν: 3279, 2925, 1733, 1554, 1474, 1399, 1262, 1132, 1033, 828, 734 cm－1; HREIMS calcd for C28H34ClN3OS [M＋H]＋ 496.2191, found 496.2189.

4-(4'-溴)苯基噻唑-2-去氢表雄酮腙(7):浅黄色固体, 产率59%. m.p. 183～185 ℃; 1H NMR (300 MHz, DMSO) δ: 0.85 (s, 3H, 18-CH3), 0.98 (s, 3H, 19-CH3), 3.33～3.21 (m, 1H, C3-αH), 4.63 (s, 1H, OH), 5.29 (d, J＝3.9 Hz, 1H, C6-H), 7.29 (s, 1H, 5-thiazole-H), 7.42 (d, J＝8.4 Hz, 2H, 3', 5'-PhH), 7.79 (d, J＝8.4 Hz, 2H, 2', 6'-PhH), 10.58 (br s, 1H, NH); 13C NMR (75 MHz, DMSO) δ: 17.3 (18-C), 19.6 (19-C), 20.7 (11-C), 23.7 (15-C), 27.4 (16-C), 31.2 (8-C), 31.4 (2-C), 31.9 (7-C), 34.4 (12-C), 36.7 (1-C), 37.4 (10-C), 42.7 (4-C), 44.3 (13-C), 50.4 (9-C), 53.7 (14-C), 70.4 (3-C), 104.6 (5-thiazole-C), 120.5 (6-C), 120.7, 128.0, 131.9, 134.7 (Ph-C), 141.9 (5-C), 149.7 (4-thiazole-C), 166.3 (17-C), 170.6 (2-thiazole-C); IR (KBr) ν: 3379, 2932, 1556, 1469, 1394, 1322, 1260, 1098, 1030, 826, 726 cm－1; HREIMS: calcd for C28H34BrN3OS [M＋H]＋ 540.1674, found 540.1684.

4.2.2 化合物8a～8b的一般合成步骤

将0.4 mmol去氢表雄酮和0.41 mmol 4-取代氨基硫脲移入50 mL圆底烧瓶中, 加入25 mL无水乙醇, 升温至60 ℃, 搅拌使其完全溶解后加入0.5 mL冰醋酸, 在60 ℃左右继续搅拌反应4 h, TLC跟踪反应至无原料点后终止反应.减压蒸出大部分溶剂, 加入20 mL蒸馏水, 采用乙酸乙酯分多次萃取.有机相依次用30 mL饱和碳酸氢钠、饱和氯化钠溶液洗涤一次, 无水硫酸钠干燥.减压蒸去溶剂, 所得到的白色固体经柱层析分离[流动相: V(乙酸乙酯):V(石油醚)＝1:2]得到目标产物.

去氢表雄酮-17-(4'-甲基)缩氨硫腙(8a):白色固体, 产率51%. m.p. 170～172 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.90 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 2.43 (dd, J＝18.0, 8.7 Hz, 1H, C4-βH), 3.22 (d, J＝4.8 Hz, 3H, NCH3), 3.49～3.59 (m, 1H, C3-αH), 3.73 (q, J＝4.8 Hz, 1H, NHCH3), 5.37 (d, J＝4.5 Hz, 1H, C6-H), 7.44 (br s, 1H, OH), 8.24 (s, 1H, NH-C＝S); 13C NMR (75 MHz, CDCl3) δ: 16.8 (18-C), 19.4 (19-C), 20.5 (11-C), 23.5 (15-C), 26.1 (16-C), 31.1 (2-C), 31.3 (8-C), 31.6 (7-C), 34.0 (NCH3), 36.6 (12-C), 37.2 (10-C), 42.2 (1-C), 44.5 (4-C), 50.3 (13-C), 53.8 (9-C), 58.4 (14-C), 71.6 (3-C), 121.0 (6-C), 141.0 (5-C), 165.9 (17-C), 178.7 (C＝S); IR (KBr) ν: 3371, 3286, 2935, 1534, 1412, 1295, 1260, 1152, 1050, 953, 821, 729 cm－1; HREIMS calcd for C21H33N3OS [M＋H]＋ 376.2421, found 376.2423.

去氢表雄酮-17-(4'-苯基)缩氨硫腙(8b):白色固体, 产率46%. m.p. 181～183 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.95 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.51 (dd, J＝18.3, 9.0 Hz, 1H, C4-βH), 3.50～3.59 (m, 1H, C3-αH), 5.39 (d, J＝4.5 Hz, 1H, C6-H), 7.26 (t, J＝7.5 Hz, 1H, Ph-H), 7.39 (t, J＝7.5 Hz, 2H, 3, 5-PhH), 7.66 (d, J＝8.1 Hz, 2H, 2, 6-Ph-H), 8.44 (s, 1H, NHPh), 9.24 (s, 1H, NHCS); 13C NMR (75 MHz, CDCl3) δ: 16.8 (18-C), 19.4 (19-C), 20.5 (11-C), 23.5 (15-C), 26.4 (16-C), 31.2 (2-C), 31.4 (7-C), 31.6 (8-C), 34.0 (12-C), 36.6 (1-C), 37.2 (10-C), 42.2 (4-C), 44.7 (13-C), 50.3 (9-C), 53.8 (14-C), 71.6 (3-C), 120.9 (6-C), 124.2, 125.9, 128.7, 138.0 (Ph-C), 141.0 (5-C), 166.7 (17-C), 176.2 (C＝S); IR (KBr) ν: 3309, 2930, 1594, 1531, 1442, 1252, 1185, 1053, 906, 729 cm－1; HREIMS calcd for C26H35N3OS [M＋H]＋ 438.2575, found 438.2579.

4.2.3 化合物9～17的一般合成步骤

称取化合物8a或8b 0.4 mmol, 4-取代-α-溴代苯乙酮0.41 mmol, 移入50 mL圆底烧瓶中, 加入25 mL无水乙醇, 升温至60 ℃, 继续搅拌反应4 h, TLC跟踪反应, 无原料点后终止反应, 减压蒸出大部分溶剂.加入20 mL蒸馏水, 乙酸乙酯萃取.合并的有机相依次用30 mL饱和碳酸氢钠、饱和氯化钠溶液洗涤一次, 无水硫酸钠干燥.减压蒸去溶剂, 得到的固体通过柱层析分离[V(乙酸乙酯):V(二氯甲烷)＝1:50]得到目标产物9～17.

去氢表雄酮N-甲基-4-苯基噻唑-2(3H)-吖嗪(9):浅黄色固体, 产率70%. m.p. 134～136 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.96 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.54～2.69 (m, 2H, C16-H), 3.29 (s, 3H, NCH3), 3.47～3.57 (m, 1H, C3-αH), 5.38 (d, J＝4.8 Hz, 1H, C6-H), 5.87 (s, 1H, 5-ArH), 7.36～7.33 (m, 2H, 3', 5'-PhH), 7.43～7.41 (m, 3H, 2', 6'-PhH, 4'-PhH); 13C NMR (75 MHz, CDCl3) δ: 17.0 (18-C), 19.5 (19-C), 20.7 (11-C), 23.5 (15-C), 27.7 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 33.4 (-NCH3), 34.4 (12-C), 36.7 (1-C), 37.3 (10-C), 42.3 (4-C), 44.1 (13-C), 50.6 (9-C), 54.1 (14-C), 71.6 (3-C), 98.8 (5-ArC), 121.2 (6-C), 128.6, 128.7, 128.9, 131.4 (Ph-C), 140.7 (4-ArC), 141.1 (5-C), 167.8 (2-ArC), 175.0 (17-C); IR (KBr) ν: 3376, 2932, 1651, 1569, 1549, 1422, 1357, 1265, 1172, 1053, 983, 836, 764, 696 cm－1; HREIMS calcd for C29H39N3OS [M＋H]＋ 476.2725, found 476.2736.

去氢表雄酮N-甲基-4-(4'-氯)苯基噻唑-2(3H)-吖嗪(10):浅黄色固体, 产率44%. m.p. 138～145 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.96 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.50～2.71 (m, 2H, C16-H), 3.27 (s, 3H, NCH3), 3.47～3.58 (m, 1H, C3-αH), 5.38 (d, J＝4.8 Hz, 1H, C6-H), 5.88 (s, 1H, 5-ArH), 7.29 (d, J＝8.4 Hz, 2H, 3', 5'-PhH), 7.41 (d, J＝8.4 Hz, 2H, 2', 6'-PhH); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.5 (15-C), 27.7 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 33.4 (-NCH3), 34.3 (12-C), 36.7 (1-C), 37.3 (10-C), 42.3 (4-C), 44.1 (13-C), 50.6 (9-C), 54.1 (14-C), 71.7 (3-C), 99.4 (5-ArC), 121.2 (6-C), 129.0, 129.8, 129.9, 135.0 (Ph-C), 139.5 (4-ArC), 141.1 (5-C), 167.5 (2-ArC), 175.2 (17-C); IR (KBr) ν: 3396, 2932, 1651, 1581, 1561, 1484, 1372, 1267, 1167, 1088, 985, 823, 731, 509 cm－1; HREIMS calcd for C29H38ClN3OS [M＋H]＋ 510.2351, found 510.2346.

去氢表雄酮N-甲基-4-(4'-硝基)苯基噻唑-2(3H)-吖嗪(11):黄色固体, 产率44%. m.p. 132～134 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.97 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 2.51～2.72 (m, 2H, C16-H), 3.33 (s, 3H, NCH3), 3.49～3.60 (m, 1H, C3-αH), 5.39 (d, J＝4.8 Hz, 1H, C6-H), 6.06 (s, 1H, 5-ArH), 7.55 (d, J＝8.7 Hz, 2H, 2', 6'-PhH), 8.31 (d, J＝8.7 Hz, 2H, 3', 5'-PhH); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.5 (15-C), 27.8 (16-C), 31.4 (2-C), 31.5 (7-C), 31.7 (8-C), 33.8 (-NCH3), 34.3 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.1 (13-C), 50.6 (9-C), 54.1 (14-C), 71.7 (3-C), 102.2 (5-ArC), 121.2 (6-C), 124.1, 129.0, 137.6, 147.8 (Ph-C), 138.6 (4-Ar-C), 141.1 (5-C), 167.2 (2-ArC), 176.0 (17-C); IR (KBr) ν: 3409, 2930, 1646, 1581, 1514, 1427, 1349, 1177, 1110, 1048, 988, 853, 756, 694 cm－1; HREIMS calcd for C29H38N4O3S [M＋H]＋ 521.2571, found 521.2586.

去氢表雄酮N-甲基-4-(4'-三氟甲基)苯基噻唑-2(3H)-吖嗪(12):黄色固体, 产率63%. m.p. 143～145 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.97 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 2.52～2.73 (m, 2H, C16-H), 3.32 (s, 3H, NCH3), 3.49～3.60 (m, 1H, C3-αH), 5.39 (d, J＝5.1 Hz, 1H, C6-H), 5.98 (s, 1H, 5-ArH), 7.50 (d, J＝8.1 Hz, 2H, 2', 6'-Ph-H), 7.71 (d, J＝8.1, 2H, 3', 5'-Ph-H); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.5 (15-C), 27.8 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 33.6 (NCH3), 34.3 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.2 (13-C), 50.6 (9-C), 54.1 (14-C), 71.7 (3-C), 100.8 (5-ArC), 121.2 (6-C), 122.0, 128.2 130.8, 131.2 (CF3), 125.7, 125.8, 128.8, 129.0, 131.2, 134.8 (Ph-C), 139.3 (4-ArC), 141.1 (5-C), 167.5 (2-ArC), 175.7 (17-C); IR (KBr) ν: 3389, 2932, 1656, 1569, 1409, 1322, 1167, 1130, 1065, 985, 851, 766, 599 cm－1; HREIMS calcd for C30H38F3N3OS [M＋H]＋544.2600, found 544.2609.

去氢表雄酮N-苯基-4-苯基噻唑-2(3H)-吖嗪(13):浅黄色固体, 产率40%. m.p. 227～229 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.94 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.40～2.55 (m, 2H, C16-H), 3.49～3.60 (m, 1H, C3-αH), 5.38 (d, J＝4.5 Hz, 1H, C6-H), 6.08 (s, 1H, 5-ArH), 7.12～7.09 (m, 2H, Ph-H), 7.18～7.31 (m, 8H, Ph-H); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.4 (15-C), 28.0 (16-C), 31.4 (2-C), 31.5 (7-C), 31.7 (8-C), 34.3 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.0 (13-C), 50.5 (9-C), 54.0 (14-C), 71.7 (3-C), 100.8 (5-ArC), 121.3 (6-C), 127.0, 128.0, 128.2, 128.3, 128.5, 131.6, 138.2 (Ph-C), 139.9 (4-ArC), 141.0 (5-C), 166.4 (2-ArC), 176.5 (17-C); IR (KBr) ν: 3311, 2927, 1646, 1544, 1491, 1352, 1197, 1050, 831, 756 cm－1; HREIMS calcd for C34H41N3OS [M＋H]＋ 538.2880, found 538.2892.

去氢表雄酮N-苯基-4-(4'-氯)苯基噻唑-2(3H)-吖嗪(14):浅黄色固体, 产率48%. m.p 207～209 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.94 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.39～2.55 (m, 2H, C16-H), 3.49～3.59 (m, 1H, C3-αH), 5.38 (d, J＝4.5 Hz, 1H, C6-H), 6.10 (s, 1H, 5-ArH), 7.03 (d, J＝8.4 Hz, 2H, Ph-H), 7.15～7.33 (m, 7H, Ph-H); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.3 (15-C), 28.0 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 34.2 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.1 (13-C), 50.5 (9-C), 54.0 (14-C), 71.7 (3-C), 101.5 (5-ArC), 121.3 (6-C), [127.2, 128.2, 128.5, 128.7, 129.2, 130.9, 134.1, 137.9] (Ph-C), 138.7 (4-ArC), 141.0 (5-C), 166.2 (2-ArC), 176.8 (17-C); IR (KBr)ν: 3426, 2932, 1646, 1561, 1486, 1349, 1297, 1195, 1093, 816, 754, 696 cm－1; HREIMS calcd for C34H40Cl-N3OS [M＋H]＋ 572.2504, found 572.2502.

去氢表雄酮N-苯基-4-(4'-硝基)苯基噻唑-2(3H)-吖嗪(15):黄色固体, 产率44%. m.p 245～247 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.94 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.40～2.55 (m, 2H, C16-H), 3.49～3.60 (m, 1H, C3-αH), 5.38 (d, J＝4.8 Hz, 1H, C6-H), 6.32 (s, 1H, 5-ArH), 7.20～7.35 (m, 7H, Ph-H), 8.05 (d, J＝9.0, 2H, Ph-H); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.3 (15-C), 28.1 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 34.2 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.1 (13-C), 50.5 (9-C), 54.0 (14-C), 71.7 (3-C), 105.0 (5-ArC), 121.2 (6-C), 123.6, 127.5, 128.0, 128.2, 128.9, 137.7, 147.0 (Ph-C), 137.8 (4-ArC), 141.0 (5-C), 165.7 (2-ArC), 177.5 (17-C); IR (KBr) ν: 3441, 2927, 1646, 1579, 1488, 1339, 1105, 1050, 853, 751 cm－1; HREIMS calcd for C34H40N4O3S [M＋H]＋ 583.2723, found 583.2743.

去氢表雄酮N-苯基-4-(4'-三氟甲基)苯基噻唑-2(3H)-吖嗪(16):黄色固体, 产率63%. m.p. 143～145 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.94 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.36～2.55 (m, 2H, C16-H), 3.49～3.60 (m, 1H, C3-αH), 5.37 (d, 1H, J＝4.5, C6-H), 6.20 (s, 1H, 5-ArH), 6.93～7.50 (m, 9H, Ph-H); 13C NMR (75MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.3 (15-C), 28.0 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 34.2 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.1 (13-C), 50.5 (9-C), 54.0 (14-C), 71.7 (3-C), 103.0 (4-ArC), 121.3 (6-C), 122.0, 125.6, 129.7, 130.1 (CF3), 125.2, 125.3, 127.3, 128.0, 128.1, 128.7, 135.0, 137.8 (Ph-C), 138.5 (4-ArC), 141.0 (5-C), 166.0 (2-ArC), 177.1 (17-C); IR (KBr) ν: 3441, 2930, 1741, 1646, 1551, 1496, 1319, 1167, 1063, 848, 749 cm－1; HREIMS calcd for C35H40F3N3OS [M＋H]＋ 606.2787, found 606. 2766.

去氢表雄酮N-苯基-4-(4'-氰基)苯基噻唑-2(3H)-吖嗪(17):黄色固体, 产率44%. m.p. 245～247 ℃; 1H NMR (300 MHz, CDCl3) δ: 0.94 (s, 3H, 18-CH3), 1.06 (s, 3H, 19-CH3), 2.39～2.55 (m, 2H, C16-H), 3.49～3.59 (m, 1H, C3-αH), 5.38 (d, J＝4.8 Hz, 1H, C6-H), 6.25 (s, 1H, 5-ArH), 7.19～7.34 (m, 7H, Ph-H), 7.48 (d, J＝8.4 Hz, 2H, Ph-H); 13C NMR (75 MHz, CDCl3) δ: 16.9 (18-C), 19.5 (19-C), 20.7 (11-C), 23.3 (15-C), 28.1 (16-C), 31.4 (2-C), 31.5 (7-C), 31.6 (8-C), 34.2 (12-C), 36.7 (1-C), 37.2 (10-C), 42.3 (4-C), 44.1 (13-C), 50.5 (9-C), 54.0 (14-C), 71.7 (3-C), 104.2 (4-ArC), 118.4 (CN), 121.2 (6-C), 111.5, 127.4, 128.0, 128.1, 128.8, 132.1, 135.9, 137.8 (Ph-C), 138.1 (4-ArC), 141.0 (5-C), 165.8 (2-ArC), 177.4 (17-C); IR (KBr) ν: 3443, 2930, 2227, 1649, 1551, 1496, 1354, 1177, 1048, 846, 749, 699 cm－1; HREIMS calcd for C35H40N4OS [M＋H]＋ 563.2842, found 563. 2845.

4.2.4 4-(4'-氨基)苯基-2-去氢表雄酮-17'-腙噻唑(18)的合成

将220 mg氯化亚锡(1 mmol)溶于0.1 mL浓盐酸中, 加入4 mL乙酸乙酯和100 mg (0.2 mmol)化合物3, 混合液逐渐变清, 然后搅拌回流2 h.待冷却至室温后加4 mL水稀释, 用1 mol/L的NaOH中和至pH值8～9之间.水层用乙酸乙酯萃取, 合并有机相, 饱和食盐水洗, 无水硫酸钠干燥.减压蒸出溶剂, 所得固体柱层析分离[洗脱剂: V(乙酸乙酯):V(二氯甲烷)＝1:10]得到深红色固体46 mg, 产率49%. m.p. 195～197 ℃; 1H NMR (300 MHz, DMSO-d6) δ: 0.85 (s, 3H, 18-CH3), 0.98 (s, 3H, 19-CH3), 3.24～3.45 (m, 1H, C3-αH), 4.62 (br s, 1H, OH), 5.29 (d, J＝3.3 Hz, 1H, C6-H), 6.56 (d, J＝8.4 Hz, 2H, 3', 5'-Ph-H), 6.77 (s, 1H, 5-ArH), 7.51 (d, J＝8.4 Hz, 2H, 2', 6'-Ph-H), 10.43 (br s, 1H, NH); 13C NMR (75 MHz, DMSO-d6) δ: 17.3 (18-C), 19.6 (19-C), 20.7 (11-C), 23.7 (15-C), 27.3 (16-C), 31.2 (8-C), 31.4 (2-C), 31.9 (7-C), 34.4 (12-C), 36.7 (1-C), 37.3 (10-C), 42.7 (4-C), 44.3 (13-C), 50.4 (9-C), 53.8 (14-C), 70.4 (3-C), 98.9 (5-thiazole-C), 120.6 (6-C), 114.2, 123.9, 126.9, 133.7 (Ph-C), 141.9 (5-C), 148.4 (4-thiazole-C), 165.6 (17-C), 170.1 (2-thiazole-C); IR (KBr) ν: 3444, 3324, 2922, 1706, 1604, 1556, 1489, 1372, 1265, 1177, 1048, 826, 736 cm－1; HREIMS calcd for C28H36N4OS [M＋H]＋ 477.2694, found 477.2688.

4.3 体外抗肿瘤活性测试

使用顺铂(Cisplatin)作为阳性对照, 采用噻唑蓝(MTT)法测试目标化合物对人宫颈癌细胞(HeLa)、人肝癌细胞(HepG)、人肺癌细胞(A549)和人肾上皮细胞(HEK-293T)四种细胞株的体外生长增殖抑制活性.所有细胞分别采用RPMI-1640培养液(加入10%牛血清和0.1 g/L青霉素G＋0.1 g/L硫酸链霉素)在37 ℃于5% CO2的潮湿气氛中培养.目标化合物以10 mg/mL的浓度溶解在无菌的DMSO (Sigma)中, 使用时根据使用浓度采用RPMI-1640培养液进行稀释.

细胞存活能力评估:将对数生长期的细胞分别以约(2～3)×104个/mL的密度接种于96孔板中, 每孔接种200 μL, 置于CO2培养箱中培养24 h.然后把不同浓度的待测化合物加入测试孔中, 每一浓度梯度设3个平行孔, 并使用等量的DMSO作为空白对照, 顺铂作为阳性对照.继续在二氧化碳培养箱中培养72 h, 细胞用无菌PBS (phosphate buffer saline)洗涤, 然后每孔加入190 µL RPMI-1640和10 μL的MTT (5 mg/ mL)溶液, 继续在二氧化碳培养箱中温育4 h; 弃去上清液, 加入200 μL DMSO, 在振动器上震荡10 min溶解形成的紫色沉淀, 随后用酶标仪在490 nm波长测定OD值.通过下式求出样品一定浓度下样品对细胞的抑制率:

抑制率(%)＝[(对照OD－空白OD)－(给药OD－空白OD)]/(对照OD－空白OD)×100%

然后以抑制率对药物浓度作图, 求出每个样品的IC50值.

辅助材料(Supporting Information) 合成化合物的1H NMR和13C NMR的谱图.这些材料可以免费从本刊网站(http://sioc-journal.cn/)上下载.

参考文献
[1] Bansal R.; Acharya P. C. Chem. Rev. , 2014, 114 : 6986. DOI:10.1021/cr4002935
[2] Mohared R. M.; Elmegged G. A.; Abdel-Salam O. M. E.; Doss S. H.; William M. G. Steroids , 2011, 76 : 1190. DOI:10.1016/j.steroids.2011.05.011
[3] Krstić N. M.; Bjelaković M. S.; Pavlović V. D.; Robeyns K.; Juranić Z. D.; Matić I.; Novaković I.; Sladić D. M. Steroids , 2012, 77 : 558. DOI:10.1016/j.steroids.2012.01.021
[4] Mohared R. M.; Al-Omran F. Steroids , 2012, 77 : 1551. DOI:10.1016/j.steroids.2012.09.007
[5] Frank E.; Schneider G. J. Steroid Biochem. Mol. Biol. , 2013, 127 : 301.
[6] Chen S.-J.; Cui J.-G.; Li Y.; Fan L. -H. Chin. J. Org. Chem. , 2011, 31 : 187. (in Chinese)
( 陈思静, 崔建国, 李莹, 范良华, 有机化学 , 2011, 31 : 187. )
[7] Zhang X.-J.; Cui J.-G.; Li Y.; Chen S.-J. Chin. J. Org. Chem. , 2010, 30 : 655. (in Chinese)
( 张晓佳, 崔建国, 李莹, 陈思静, 有机化学 , 2010, 30 : 655. )
[8] Traf D. T. P. Cancer Lett. , 2006, 243 : 202. DOI:10.1016/j.canlet.2005.11.034
[9] Cui J.-G.; Liu L.; Gan C.-F.; Xiao Q. Prog. Chem. , 2014, 2/3 : 655–320. (in Chinese)
( 崔建国, 刘亮, 甘春芳, 肖燕敏, 化学进展 , 2014, 2/3 : 655–320. )
[10] Cui S.-F.; Wang Y.; Lü J.-S.; Damu Guri L. V.; Zhou C.-H. Sci. China, Ser B , 2012, 42 : 1105. (in Chinese)
( 崔胜峰, 王艳, 吕敬松, Damu GuriL. V., 周成合, 中国科学, 化学 , 2012, 42 : 1105. )
[11] Lesyk R.; Zimenkovsky B.; Atamanyuk D.; Jensen F.; Kononowicz K. K.; Gzella A. Bioorg. Med. Chem. , 2006, 14 : 5230. DOI:10.1016/j.bmc.2006.03.053
[12] Kashfi K. Adv. Pharmacol. , 2009, 57 : 31. DOI:10.1016/S1054-3589(08)57002-5
[13] Ramla M. M.; Omar M. A.; El-Khamry A. M.; El-Diwan H. I. Bioorg. Med. Chem. , 2006, 14 : 7324. DOI:10.1016/j.bmc.2006.06.033
[14] Sun L. P.; Jiang Z.; Gao L. X.; Liu X. F.; Quan Y. C.; Zheng G. H.; Li J.; Piao H. Chin. J. Org. Chem. , 2013, 33 : 1496. (in Chinese)
( 孙良鹏, 姜哲, 高立信, 刘晓芳, 全迎春, 郑光浩, 李佳, 朴虎日, 有机化学 , 2013, 33 : 1496. DOI:10.6023/cjoc201212012 )
[15] Mohareb R. M.; Al-Omran F. Steroids , 2012, 77 : 1551. DOI:10.1016/j.steroids.2012.09.007
[16] Yuan M.-Y.; Liu M.; Zhang Y.-Q.; Yan H.-F.; Li D.-F.; Zhang D. N.; Liu H. M. Chin. J. Org. Chem. , 2012, 32 : 1746. (in Chinese)
( 袁明月, 刘敏, 张英群, 闫红飞, 李德富, 张冬暖, 刘卉闵, 有机化学 , 2012, 32 : 1746. DOI:10.6023/cjoc1201302 )
[17] Chen C.-J.; Song B. A. Pesticide , 2005, 44(2) : 53. (in Chinese)
( 陈才俊, 宋宝安, 农药 , 2005, 44 (2) : 53. )
[18] Shingate B. B.; Hazra B. G.; Salunke D. B.; Pore V. S.; Shirazi F.; Deshpande M. V. Eur. J. Med. Chem. , 2011, 46 : 3681. DOI:10.1016/j.ejmech.2011.05.032
[19] Cui J.; Qi B.; Gan C.; Liu Z.; Huang H.; Lin Q.; Zhao D.; Huang. Y. Mar. Drugs , 2015, 13 : 2488. DOI:10.3390/md13042488
[20] Cui J.; Liu L.; Zhao D.; Gan C.; Huang X.; Xiao Q.; Qi B.; Yang L.; Huang Y. Steroids , 2015, 95 : 32.
[21] Zhao D.-D.; Qi B.-B.; Yang L.; Huang Y.-M.; Gan C.-F.; Cui J.-G. Chin. J. Med. Chem. , 2014, 24 : 470. (in Chinese)
( 赵丹丹, 戚斌斌, 杨雷, 黄燕敏, 甘春芳, 崔建国, 中国药物化学杂志 , 2014, 24 : 470. )
[22] http://www.cnki.com.cn/article/cjfdtotal-hxyj201505020.htmQi, B.-B.; Liu, L.; Zhao, D.-D; Yang, L.; Huang, Y. M.; Cui, J.-G. Chem. Res. Appl. 2015, 27, 665 (in Chinese).
(戚斌斌, 刘亮, 赵丹丹, 杨雷, 甘春芳, 黄燕敏, 崔建国, 化学研究与应用, 2015, 27, 665.)
[23] Yang S.; Fu Z.-D.; Han R. Acta Pharm. Sinica , 2001, 36 : 576. (in Chinese)
( 杨笋, 付招娣, 韩锐, 药学学报 , 2001, 36 : 576. )
[24] Ahluwalia V. K.; Chibber S. S.; Goyal B. Indian J. Chem.. Sect. B , 1996, 35 : 856.
[25] Yuan M.-Y.; Liu W. H.; Zhang Y. Q.; Yan H. F.; Zhang D.-N.; Liu H.-M.; Wang J.-P. Chin. J. Org. Chem. , 2013, 33 : 1108. (in Chinese)
( 袁明月, 刘伟华, 张英群, 闫红飞, 张冬暖, 刘卉闵, 王建平, 有机化学 , 2013, 33 : 1108. DOI:10.6023/cjoc201210019 )
X