2-取代苯氨基喹唑啉衍生物的合成及抗肿瘤活性研究

引用本文: 潘晓乐, 王磊, 顿艳艳, 方浩. 2-取代苯氨基喹唑啉衍生物的合成及抗肿瘤活性研究[J]. 有机化学, 2016, 36(5): 1044-1050. doi: 10.6023/cjoc201510018 shu

Citation: Pan Xiaole, Wang Lei, Dun Yanyan, Fang Hao. Synthesis and Antiproliferative Activity Studies on 2-Substitued Aniline Quinazoline Derivatives[J]. Chinese Journal of Organic Chemistry, 2016, 36(5): 1044-1050. doi: 10.6023/cjoc201510018 shu

2-取代苯氨基喹唑啉衍生物的合成及抗肿瘤活性研究

1.
山东大学药学院济南 250012

通讯作者: 方浩,E-mail: haofangcn@sdu.edu.cn

基金项目:
国家自然科学基金 No.81373281

摘要: 以2,4-喹唑啉二酮为起始原料,通过氯代、Suzuki偶联和芳环亲核取代反应得到一系列2-取代苯氨基喹唑啉衍生物,并经¹H NMR,¹³C NMR和HRMS进行结构确证.应用溴化噻唑蓝四氮唑(MTT)法对目标化合物进行了初步体外抗肿瘤细胞增殖活性研究.结果表明,部分化合物具有较好的体外抗肿瘤细胞增殖活性,其中活性化合物4u和4v对Hela,A549和MCF-7三种肿瘤细胞株的增殖均有明显抑制作用.

关键词:

English

Synthesis and Antiproliferative Activity Studies on 2-Substitued Aniline Quinazoline Derivatives

1.
Department of Medicinal Chemistry, School of Pharmaceutical Sciences, Shandong University, Ji'nan 250012

Corresponding author: Fang Hao, haofangcn@sdu.edu.cn

Received Date: 16 October 2015
Revised Date: 03 January 2016
Fund Project:
Project supported by the National Natural Science Foundation of China

Abstract: A series of 2-substituted aniline quinazoline derivatives were prepared with the starting material of quinazoline-2,4(1H,3H)-dione, and then followed by the reaction of chlorination, Suzuki reaction and nucleophilic substitution. The structures of target compounds were identified by ¹H NMR, ¹³C NMR and HRMS. The preliminary biological evaluations were performed using thiazolyl blue tetrazolium bromide (MTT) method to test their antiproliferative activities against some tumor cell lines. The results suggest that some compounds have the abilities to inhibit the growth of Hela tumor cell lines, and most potent compounds 4u and 4v alsoexhibit good antiproliferative activities against A549 and MCF-7 tumor cell lines.

Key words:

喹唑啉类化合物最早从Gabriel植物中分离提取得到,随后发现其具有镇静^{[1, 2]}、抗炎^[3]、抗高血压^[4～6]、抗疟^[7]和抗真菌^[8～10]等多种药理作用. 近年来,喹唑啉骨架作为药效活性片段,已在抗肿瘤药物的研发中不断被成功应用^[11～13]. 到目前为止,已有Albaconazole^{[14, 15]}、Gefitinib^[16]、Erlotinib^[17]、Vandetanib^[18]等多个喹唑啉衍生物结构作为抗肿瘤药物上市,被应用于肺癌和胰腺癌等恶性肿瘤的临床治疗^[19～21].

本课题组近期曾针对国外Ⅱ期临床研究的药物R-Roscovitine进行结构改造^[22]. 前期针对该药的构效关系研究主要基于其嘌呤环(嘧啶并咪唑)母核,在嘌呤环2位和4位引入不同的芳胺或脂肪胺片断^[23]. 根据药物设计学原理,我们在保留R-Roscovitine母核嘧啶环的基础上,将咪唑环以苯环替代,设计合成了一系列2-取代芳胺喹唑啉类衍生物,并进行构效关系研究. 合成路线设计见Scheme 1.

图图式1 目标化合物的合成 Figure 图式1. Synthetic route of the target compounds

图选项

下载全尺寸图像

下载幻灯片

表 1 化合物4的结构 Table 1. The structure of the compounds 4

表选项

导出Excel

下载全尺寸表图像

Compd.	R¹	R²
4a	Ph	H
4b	Ph	4-CH₃
4c	Ph	3-NO₂
4d	Ph	3-Morpholino
4e	Ph	4-Morpholino
4f	Ph	4-SO₂NH₂
4g	4-CH₃C₆H₄	4-CH₃
4h	4-CH₃C₆H₄	3-NO₂
4i	4-CH₃C₆H₄	4-Morpholino
4j	3-NO₂C₆H₄	H
4k	3-NO₂C₆H₄	4-CH₃
4l	3-NO₂C₆H₄	3-NO₂
4m	3-NO₂C₆H₄	4-Morpholino
4n	Thiophen-3-yl	H
4o	Thiophen-3-yl	4-CH₃
4p	Thiophen-3-yl	3-NO₂
4q	Thiophen-3-yl	4-Morpholino
4r	Thiophen-3-yl	4-CH₃
4s	Thiophen-3-yl	3-NO₂
4t	Thiophen-3-yl	4-Morpholino
4u	Furan-2-yl	H
4v	Furan-2-yl	4-CH₃
4w	Furan-2-yl	3-NO₂
4x	Furan-2-yl	4-Morpholino

表 1 化合物4的结构
Table 1. The structure of the compounds 4

1 结果与讨论

1.1 目标化合物的合成

化合物4a～4x的合成是以2,4-喹唑啉二酮为起始原料,首先经过三氯氧磷的处理得到2,4-二氯喹唑啉(2),然后与各种芳香硼酸发生Suzuki偶联得关键中间体3a～3f,最后在N₂的保护下或者微波条件下与各种取代的苯胺反应得到终产物4a～4x. 目标化合物的结构均经¹H NMR,¹³C NMR和HRMS确证.

1.2 目标化合物的抗肿瘤活性

所有目标化合物浓度在50 μmol/L条件下,应用Hela肿瘤细胞株对其进行体外抗增殖活性测试,结果见图 1. 实验结果说明,喹唑啉C(4)位引入3-硝基苯基(如化合物4j、4k、4l和4m)、3-噻吩基(如化合物4o、4p和4q)及2-噻吩基(如化合物4r、4s和4t)的活性较差; 喹唑啉C(2)位的苯胺若带有硝基或者吗啉基(如化合物4h、4m、4p、4w和4x)不利于提高其对肿瘤细胞的抑制作用,而在喹唑啉的C(4)位引入2-呋喃基、C(2)位引入苯胺和对甲基苯胺时(如化合物4u和4v),目标化合物的抗肿瘤活性得到明显改善.

图 1 目标化合物浓度为50 μmol/L时对Hela细胞的抑制作用 Figure 1. Inhibitions of all the target compounds to Hela cell line at 50 μmol/L

图选项

下载全尺寸图像

下载幻灯片

我们选取活性化合物4u和4v(抑制率大于80%)并测试其对Hela、A549和MCF-7三种肿瘤细胞株的抗增殖效果(Table 2). 结果显示,两个活性化合物抑制Hela、A549肿瘤细胞株的活性与阳性对照药R-Roscovitine相当或优于阳性对照药R-Roscovitine,对MCF-7肿瘤细胞株的抑制作用不如阳性对照药R-Roscovitine. 分析两个活性化合物对不同瘤株的抗增殖效果,可发现它们对于Hela肿瘤细胞的抑制作用明显好于其它两株肿瘤细胞A549和MCF-7.

表 2 代表化合物的体外抗增殖活性 Table 2. Antiproliferative activities of representative compounds

表选项

导出Excel

下载全尺寸表图像

Compd.	IC₅₀/(μmol•L^-1)
Compd.	Hela	A549	MCF-7
4u	12.76±2.36	32.95±1.17	16.21±2.00
4v	14.93±1.59	30.47±1.56	26.24±0.76
R-Roscovitine	14.01±2.62	78.70±6.03	12.72±1.25

表 2 代表化合物的体外抗增殖活性
Table 2. Antiproliferative activities of representative compounds

2 结论

本文以R-Roscovitine为先导化合物并结合喹唑啉骨架在肿瘤药物研发的应用,根据药物设计学原理,设计并合成了24个结构全新的目标化合物. 所有目标化合物的结构均经¹H NMR,¹³C NMR和HRMS确证. 在体外抗肿瘤细胞增殖活性的试验中,有多个化合物在浓度为50 μmol/L时对Hela肿瘤细胞株都有明显的抑制作用; 其中初筛时活性最好的化合物4u、4v对Hela、A549两株肿瘤细胞抗增殖作用与阳性对照药R-Roscovitine相当或优于R-Roscovitine,对MCF-7肿瘤细胞株的抗增殖作用不如阳性对照药R-Roscovitine. 上述结果为今后研究新型喹唑啉类抗肿瘤药物奠定了良好基础.

3 实验部分

3.1 仪器与试剂

合成中所用到的原料均为市售的国产试剂,纯度为96%～99%; 反应溶剂为分析纯,未经处理直接使用. 人子宫颈癌细胞Hela、人乳腺癌细胞MCF-7、人肺腺癌细胞A549; 改良型RPMI1640培养基(美国Hyclone公司),2.5 g/L胰蛋白酶(美国Gibco公司),10%(体积分数)胚牛血清(美国Hyclone公司). 阳性对照药R-Roscovatine购自MedChemExpress (MCE)中国分公司.

Bruker-300 MHz和Bruker-400 MHz核磁共振波谱仪; Agilent 6520型高分辨电喷雾电离质谱仪; RY-1型熔点仪(天津市分析仪器厂); Thermo Varioskan Flash全波长多功能酶标仪; Thermo Forma 3111型二氧化碳恒温培养箱; Olympus CKX31双目倒置显微镜.

3.2 化合物的合成

3.3 抗肿瘤活性实验

采用四甲基偶氮唑盐(MTT)法对24个目标化合物进行体外抗肿瘤活性评价. 测试细胞为Hela、A549和MCF-7.

将处于对数生长期的肿瘤细胞,用含10%(体积分数)胚牛血清的RPMI1640培养液调整细胞数(所选细胞先用2.5×10^-3 g/mL胰蛋白酶消化后调整为细胞数4×10⁴ cell/mL的细胞悬液),接种于96孔培养板中,37 ℃、5% CO₂条件下培养8 h后,加入不同浓度的待测化合物,继续培养48 h. 然后每孔加入10 μL MTT (5 mg/mL),继续培养4 h后,弃去孔中液体,每孔加入150 μL DMSO,37 ℃恒温振摇5～10 min,检测570 nm波长处的OD值,计算所测化合物对细胞的抑制率以及IC₅₀值.

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

3.2.2 目标化合物4a～4x的合成

将化合物3 (1 mmol)与各种取代的苯胺(1 mmol)混合,加入正丁醇或二甲亚砜(4～6 mL),在氮气保护下110～120 ℃反应6～8 h(或反应混合物微波加热110～150 ℃反应15～45 min),TLC监测至反应结束. 反应液冷至室温,静置12 h后过滤,滤渣经柱层析得到目标化合物4a～4x.

N,4-二苯基喹唑啉-2-胺(4a): 黄色固体,产率75%. m.p. 133～134 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 9.93 (s,1H,NH),8.04 (d,J=7.8 Hz,2H,ArH),7.85～7.74 (m,5H,ArH),7.65～7.61 (m,3H,ArH),7.38～7.31 (m,3H,ArH),6.98 (t,J=7.2 Hz,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 169.80,156.78,152.63,141.19,137.11,134.60,130.31,129.96,128.98,128.90,127.45,126.74,124.05,121.93,119.22,118.78; HRMS (AP-ESI) calcd for C₂₀H₁₆N₃ [M+H]⁺ 298.1344,found 298.1335.

4-苯基-N-(4-甲基苯基)喹唑啉-2-胺(4b): 黄色固体,产率55%. m.p. 137～139 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.86 (s,1H,NH),7.90 (d,J=8.4 Hz,2H,ArH),7.83～7.72 (m,5H,ArH),7.63～7.61 (m,3H,ArH),7.35～7.31 (m,1H,ArH),7.15 (d,J=8.0 Hz,2H,ArH),2.28 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ:169.81,156.79,152.56,138.55,137.14,134.57,130.84,130.30,129.94,129.40,128.89,127.46,126.55,123.87,119.44,118.68,20.92; HRMS (AP-ESI) calcd for C₂₁H₁₈N₃ [M+H]⁺ 312.1501,found 312.1495.

N-(3-硝基苯基)-4-苯基喹唑啉-2-胺(4c): 黄色固体,产率89%. m.p. 166～168 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 10.54 (s,1H,NH),9.21 (s,1H,ArH),8.32 (d,J=8.0 Hz,1H,ArH),7.92～7.88 (m,2H,ArH),7.85～7.80 (m,4H,ArH),7.65～7.60 (m,4H,ArH),7.46 (t,J=8.0 Hz,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 170.54,155.93,151.31,148.61,142.09,136.75,135.20,130.69,130.25,130.14,128.99,127.71,126.19,125.38,125.03,118.99,116.49,113.32; HRMS (AP-ESI) calcd for C₂₀H₁₅N₄O₂ [M+H]⁺ 343.1195,found 343.1192.

N-(3-吗啉基苯基)-4-苯基喹唑啉-2-胺(4d): 红色固体,产率73%. m.p. 154～156 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 10.16 (s,1H,NH),8.04 (s,1H,ArH),7.89～7.78 (m,5H,ArH),7.65～7.61 (m,3H,ArH),7.45～7.38 (m,2H,ArH),7.30 (t,J=8.0 Hz,1H,ArH),6.85 (s,1H,ArH),3.87 (br s,4H,O(CH₂)₂),3.26 (br s,4H,N(CH₂)₂); ¹³C NMR (100 MHz,DMSO-d₆) δ: 171.74,154.95,148.92,140.69,136.53,135.43,130.86,130.07,129.94,128.86,127.96,124.86,118.30,112.30,109.53,65.54,51.47; HRMS (AP-ESI) calcd for C₂₄H₂₃N₄O [M+H]⁺ 383.1872,found 383.1866.

N-(4-吗啉基苯基)-4-苯基喹唑啉-2-胺(4e): 黄色固体,产率70%. m.p. 152～154 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.72 (s,1H,NH),7.87 (d,J=8.8 Hz,2H,ArH),7.80～7.74 (m,4H,ArH),7.69 (d,J=8.4 Hz,1H,ArH),7.62～7.61 (m,3H,ArH),7.31 (t,J=8.0 Hz,1H,ArH),6.96 (d,J=8.8 Hz,2H,ArH),3.76 (d,J=4.4 Hz,4H,O(CH₂)₂),3.07 (d,J=4.4 Hz,4H,N(CH₂)₂). ¹³C NMR (100 MHz,DMSO-d₆) δ: 169.09,156.41,152.38,146.15,136.71,133.97,133.13,129.72,129.40,128.37,126.93,126.04,123.02,119.96,118.09,115.62,66.17,49.27; HRMS (AP-ESI) calcd for C₂₄H₂₃N₄O [M+H]⁺ 383.1872,found 383.1861.

4-[(4-苯基喹唑啉-2-基)胺基]苯磺酰胺(4f): 黄色固体,产率64%. m.p. 246～248 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 10.38 (s,1H,NH),8.20 (d,J=8.8 Hz,2H,ArH),7.90～7.77 (m,7H,ArH),7.65～7.63 (m,3H,ArH),7.44～7.40 (m,1H,ArH),7.22 (s,2H,SO₂NH₂); ¹³C NMR (100 MHz,DMSO-d₆) δ: 169.59,155.85,151.79,143.75,136.43,136.27,134.34,129.98,129.55,128.47,127.04,126.56,126.47,124.29,118.60,117.92; HRMS (AP-ESI) calcd for C₂₀H₁₇N₄O₂S [M+H]⁺ 377.1072,found 377.1061.

N,4-二(4-甲基苯基)喹唑啉-2-胺(4g): 黄色固体,产率76%. m.p. 218～220 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 10.01 (s,1H,NH),7.86～7.80 (m,4H,ArH),7.75～7.66 (m,3H,ArH),7.45 (d,J=8.1 Hz,2H,ArH),7.38 (t,J=7.8 Hz,1H,ArH),7.17 (d,J=8.4 Hz,2H,ArH),2.45 (s,3H,CH₃),2.29 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 172.80,154.39,147.05,141.38,136.15,133.61,133.47,130.37,129.81,129.69,128.50,125.15,122.60,121.58,118.18,21.53,21.00; HRMS (AP-ESI) calcd for C₂₂H₂₀N₃ [M+H]⁺ 326.1657,found 326.1653.

N-(3-硝基苯基)-4-(4-甲基苯基)喹唑啉-2-胺(4h): 黄色固体,产率51%. m.p. 201～203 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 10.48 (s,1H,NH),9.21 (s,1H,ArH),8.32 (dd,J=8.1,1.2 Hz,1H,ArH),7.93～7.86 (m,2H,ArH),7.84～7.79 (m,2H,ArH),7.74 (d,J=8.1 Hz,2H,ArH),7.64 (t,J=8.1 Hz,1H,ArH),7.46～7.40 (m,3H,ArH),2.46 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 170.44,156.01,151.44,148.64,142.18,140.57,135.14,134.00,130.29,130.22,129.59,127.80,126.25,125.38,124.99,119.04,116.46,113.30,21.50; HRMS (AP-ESI) calcd for C₂₁H₁₇N₄O₂ [M+H]⁺ 357.1352,found 357.1354.

N-(4-吗啉基苯基)-4-(4-甲基苯基)喹唑啉-2-胺(4i): 黄色固体,产率18%. m.p. 159～161 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.69 (s,1H,NH),7.87 (d,J=8.8 Hz,2H,ArH),7.80～7.77 (m,2H,ArH),7.67～7.65 (m,3H,ArH),7.43 (d,J=8 Hz,2H,ArH),7.30 (t,J=8 Hz,1H,ArH),6.95 (d,J=9.2 Hz,2H,ArH),3.76 (t,J=4.4 Hz,4H,O(CH₂)₂),3.07 (t,J=4.4 Hz,4H,N(CH₂)₂),2.44 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 169.51,156.95,152.91,146.63,139.95,134.43,134.38,133.68,129.95,129.45,127.48,126.52,123.44,120.47,118.63,116.12,66.67,49.79,21.46; HRMS (AP-ESI) calcd for C₂₅H₂₅N₄O [M+H]⁺ 397.2028,found 397.2025.

4-(3-硝基苯基)-N-苯基喹唑啉-2-胺(4j): 黄色固体,产率71%. m.p. 178～179 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 10.03 (s,1H,NH),8.60 (t,J=2.1 Hz,1H,ArH),8.50～8.46 (m,1H,ArH),8.27 (dt,J=7.5,0.9 Hz,1H,ArH),8.04 (d,J=7.8 Hz,2H,ArH),7.95～7.78 (m,4H,ArH),7.41～7.32 (m,3H,ArH),7.02 (t,J=7.5 Hz,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 167.43,156.64,152.77,148.32,141.03,138.50,136.48,134.94,130.64,128.99,126.99,126.88,125.03,124.72,124.41,122.08,119.27,118.51; HRMS (AP-ESI) calcd for C₂₀H₁₅N₄O₂ [M+H]⁺ 343.1195,found 343 .1194.

4-(3-硝基苯基)-N-(4-甲基苯基)喹唑啉-2-胺(4k): 红色固体,产率56%. m.p. 184～186 ℃; ¹H NMR (400 MHz,CDCl₃) δ: 11.25 (brs,1H,NH),8.67 (s,1H,ArH),8.55 (d,J=7.6 Hz,1H,ArH),8.15 (d,J=6.8 Hz,1H,ArH),7.99～7.94 (m,3H,ArH),7.86 (t,J=7.6 Hz,1H,ArH),7.55 (d,J=7.6 Hz,3H,ArH),7.22 (d,J=7.6 Hz,2H,ArH),2.36 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 169.16,155.38,149.53,148.31,138.07,136.99,136.49,135.86,132.55,130.79,129.66,127.55,125.43,124.95,124.81,124.49,120.68,118.24,20.96; HRMS (AP-ESI) calcd for C₂₁H₁₇N₄O₂ [M+H]⁺357.1352,found 357.1344.

N,4-二(-3-硝基苯基)喹唑啉-2-胺(4l): 黄色固体,产率26%. m.p. 236～238 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 10.59 (s,1H,NH),9.20 (t,J=2.1 Hz,1H,ArH),8.64 (t,J=2.1 Hz,1H,ArH),8.52～8.48 (m,1H,ArH),8.34～8.27 (m,2H,ArH),7.97～7.84 (m,5H,ArH),7.66 (t,J=8.4 Hz,1H,ArH),7.50～7.44 (m,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 167.80,156.23,152.36,148.66,148.38,142.33,138.32,136.57,135.32,130.72,130.29,127.16,127.02,125.21,125.08,124.84,118.91,116.32,112.96; HRMS (AP-ESI) calcd for C₂₀H₁₄N₅O₄ [M+H] ⁺ 388.1046,found 388.1040.

N-(4-吗啉基苯基)-4-(3-硝基苯基)喹唑啉-2-胺(4m): 红色固体,产率19%. m.p. 144～146 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.83 (s,1H,NH),8.58 (t,J=2.0 Hz,1H,ArH),8.49 (dd,J=8.0,1.2 Hz,1H,ArH),8.25 (d,J=8 Hz,1H,ArH),7.93 (t,J=8 Hz,1H,ArH),7.87～7.84 (m,2H,ArH),7.82～7.80 (m,1H,ArH),7.79 (d,J=8.4 Hz,1H,ArH),7.72 (d,J=8 Hz,1H,ArH),7.35 (t,J=7.2 Hz,1H,ArH),6.97 (d,J=8.8 Hz,2H,ArH),3.77 (t,J=4.8 Hz,4H,O(CH₂)₂),3.08 (t,J=4.8 Hz,4H,N(CH₂)₂); ¹³C NMR (100 MHz,DMSO-d₆) δ: 167.24,156.78,153.04,148.32,146.77,138.62,136.44,134.82,133.44,130.61,126.97,126.68,124.96,124.67,123.88,120.52,118.32,116.09,66.66,49.73; HRMS (AP-ESI) calcd for C₂₄H₂₂N₅O₃ [M+H]⁺ 428.1723,found 428.1714.

N-苯基-4-(噻吩-3-基)喹唑啉-2-胺(4n): 黄色固体,产率50%. m.p. 126～128 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.84 (s,1H,NH),8.22 (dd,J=3.2,1.2 Hz,1H,ArH),8.09 (d,J=8.0 Hz,1H,ArH),8.02 (d,J=8.0 Hz,2H,ArH),7.84～7.80 (m,2H,ArH),7.74 (d,J=8.0 Hz,1H,ArH),7.66 (dd,J=5.2,1.2 Hz,1H,ArH),7.40～7.31 (m,3H,ArH),6.70 (t,J=7.2 Hz,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 164.51,156.85,152.76,141.22,138.42,134.55,129.60,129.41,128.97,127.37,127 .25,126.71,124.14,121.89,119.26,118.83; HRMS (AP-ESI) calcd for C₁₈H₁₄N₃S [M+H]⁺ 304.0908,found 304.0901.

4-(噻吩-3-基)-N-(4-甲基苯基)喹唑啉-2-胺(4o): 砖红色固体,产率54%. m.p. 224～226 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 9.91 (s,1H,NH),8.25～8.24 (m,1H,ArH),8.10 (d,J=8.1 Hz,1H,ArH),7.86～7.80 (m,4H,ArH),7.73 (d,J=8.1 Hz,1H,ArH),7.65 (dd,J=5.1,1.2 Hz,1H,ArH),7.42 (t,J=7.8 Hz,1H,ArH),7.17 (d,J=8.4 Hz,2H,ArH),2.29 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 167.17,154.19,137.75,136.35,135.88,133.68,132.39,129.85,129.41,128.28,128.05,125.46,121.74,118.01,21.02; HRMS (AP-ESI) calcd for C₁₉H₁₆N₃S [M+H]⁺ 318.1065,found 318.1050.

N-(3-硝基苯基)-4-(噻吩-3-基)喹唑啉-2-胺(4p): 黄色固体,产率89%. m.p. 205～207 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 10.99 (s,1H,NH),9.77 (t,J=2.1 Hz,1H,ArH),8.87～8.84 (m,2H,ArH),8.74 (d,J=8.4 Hz,1H,ArH),8.49～8.34 (m,4H,ArH),8.27 (dd,J=5.4,1.2 Hz,1H,ArH),8.21 (t,J=8.1 Hz,1H,ArH),8.06～8.01 (m,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 165.16,155.84,151.16,148.59,142.01,138.09,135.23,130.65,130.24,129.37,127.65,127.54,125.92,125.48,125.18,118.92,116.53,113.43; HRMS (AP-ESI) calcd for C₁₈H₁₃N₄O₂S [M+H]⁺ 349.0759,found 349.0751.

N-(4-吗啉基苯基)-4-(噻吩-3-基)喹唑啉-2-胺(4q): 黄色固体,产率8%. m.p. 210～212 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.69 (s,1H,NH),8.25 (dd,J=2.4,0.8 Hz,1H,ArH),8.09 (d,J=8 Hz,1H,ArH),7.91 (d,J=8.8 Hz,2H,ArH),7.86～7.81 (m,2H,ArH),7.72～7.67 (m,2H,ArH),7.40 (t,J=0.8 Hz,1H,ArH),7.01 (d,J=9.2 Hz,2H,ArH),3.81 (t,J=4.4 Hz,4H,O(CH₂)₂), 3.12 (t,J=4.4 Hz,4H,N(CH₂)₂); ¹³C NMR (100 MHz,DMSO-d₆) δ: 164.35,156.98,152.99,146.64,138.51,134.45,133.66,129.43,129.40,127.32,127.25,126.51,123.63,120.50,118.65,116.13,66.67,49.78; HRMS (AP-ESI) calcd for C₂₂H₂₁N₄OS [M+H]⁺ 389.1436,found 389.1430.

4-(噻吩-2-基)-N-(4-甲基苯基)喹唑啉-2-胺(4r): 红色固体,产率59%. m.p. 206～208 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 10.13 (s,1H,NH),8.41 (d,J=8 Hz,1H,ArH),8.09 (d,J=3.6 Hz,1H,ArH),8.05 (d,J=5.2 Hz,1H,ArH),7.90～7.87 (m,1H,ArH),7.77～7.73 (m,3H,ArH),7.49 (t,J=14.8 Hz,1H,ArH),7.39 (dd,J=4.8,3.6 Hz,1H,ArH),7.21 (d,J=8.4 Hz,2H,ArH),2.31 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 164.64,153.06,145.47,140.21,136.78,135.47,134.97,133.98,129.93,129.82,127.98,125.95,121 .85,121.35,116.70,21.02; HRMS (AP-ESI) calcd for C₁₉H₁₆N₃S [M+H]⁺ 318.1065,found 318.1058.

N-(3-硝基苯基)-4-(噻吩-2-基)喹唑啉-2-胺(4s): 黄色固体,产率16%. m.p. 169～171 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 10.37 (s,1H,NH),9.18 (t,J=2.4 Hz,1H,ArH),8.43 (d,J=8 Hz,1H,ArH),8.30 (dd,J=8.4,1.6 Hz,1H,ArH),8.07 (dd,J=3.6,0.8 Hz,1H,ArH),8.03 (dd,J=5.2,1.2 Hz,1H,ArH),7.93 (m,1H,ArH),7.85 (dd,J=8,1.6 Hz,1H,ArH),7.79 (d,J=8 Hz,1H,ArH),7.64 (t,J=8 Hz,1H,ArH),7.53～7.49 (m,1H,ArH),7.39～7.37 (m,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 161.74,156.05,152.89,148.62,142.46,140.56,134.93,132.54,131.96,130.12,129.16,126.99,126.94,125.19,125.11,117.93,116.08,113.03; HRMS (AP-ESI) calcd for C₁₈H₁₃N₄O₂S [M+H]⁺ 349.0759,found 349.0757.

N-(4-吗啉基苯基)-4-(噻吩-2-基)喹唑啉-2-胺(4t): 橙色固体,产率36%. m.p. 229～231 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.57 (s,1H,NH),8.32 (d,J=8.4 Hz,1H,ArH),7.99 (d,J=3.6 Hz,1H,ArH),7.96 (d,J=5.2 Hz,1H,ArH),7.84～7.78 (m,3H,ArH),7.66 (d,J=8 Hz,1H,ArH),7.39～7.33 (m,2H,ArH),6.96 (d,J=8.8 Hz,2H,ArH),3.76 (t,J=4.4 Hz,4H,O(CH₂)₂),3.07 (t,J=4.4 Hz,4H,N(CH₂)₂); ¹³C NMR (100 MHz,DMSO-d₆) δ: 161.37,156.68,153.53,146.69,140.88,134.59,133.54,132.01,131.42,129.01,126.83,126.69,123.96,120.64,117.47,116.08,66.67,49.76; HRMS (AP-ESI) calcd for C₂₂H₂₁N₄OS [M+H]⁺ 389.1436,found 389.1431.

4-(呋喃-2-基)-N-苯基喹唑啉-2-胺(4u): 黄色固体,产率14%. m.p. 126～128 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.84 (s,1H,NH),8.62 (d,J=8 Hz,1H,ArH),8.17 (d,J=1.2 Hz,1H,ArH),8.02 (d,J=8 Hz,2H,ArH),7.85～7.81 (m,1H,ArH),7.72 (d,J=8.4 Hz,1H,ArH),7.57 (d,J=3.2 Hz,1H,ArH),7.45～7.41 (m,1H,ArH),7.36 (t,J=7.6 Hz,2H,ArH),7.01 (t,J=7.6 Hz,1H,ArH),6.88～6.86 (m,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 156.7 9,156.43,153.42,152.23,147.25,141.23,134.61,128.98,126.92,126.89,124.41,121.86,119.22,116.89,116.60,113.07; HRMS (AP-ESI) calcd for C₁₈H₁₄N₃O [M+H]⁺ 288.1137,found 288.1134.

4-(呋喃-2-基)-N-(4-甲基苯基)喹唑啉-2-胺(4v): 黄色固体,产率60%. m.p. 124～126 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.83 (s,1H,NH),8.61 (d,J=8 Hz,1H,ArH),8.17 (s,1H,ArH),7.85～7.80 (m,3H,ArH),7.69 (d,J=8.4 Hz,1H,ArH),7.56 (d,J=3.2 Hz,1H,ArH),7.42 (t,J=7.5 Hz,1H,ArH),7.16 (d,J=7.6 Hz,2H,ArH),6.87 (dd,J=3.2,1.5 Hz,1H,ArH),2.29 (s,3H,CH₃); ¹³C NMR (100 MHz,DMSO-d₆) δ: 157.70,153.43,151.24,148.75,135.73,135.39,132.77,129.20,127.14,124.96,121.76,120.82,119.37,115.47,113.25,20.43; HRMS (AP-ESI) calcd for C₁₉H₁₆N₃O [M+H]⁺ 302.1293,found 302.1285.

4-(呋喃-2-基)-N-(3-硝基苯基)喹唑啉-2-胺(4w): 黄色固体,产率22%. m.p. 180 (dec.) ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 10.40 (s,1H,NH),9.23 (t,J=2 Hz,1H,ArH),8.70 (d,J=8 Hz,1H,ArH),8.26 (d,J=7.6 Hz,1H,ArH),8.20 (d,J=0.8 Hz,1H,ArH),7.91～7.87 (m,1H,ArH),7,84 (dd,J=8,1.6 Hz,1H,ArH),7.77 (d,J=8.4 Hz,1H,ArH),7.66～7.60 (m,2H,ArH),7.53 (t,J=8 Hz,1H,ArH),6.91～6.89 (m,1H,ArH); ¹³C NMR (100 MHz,DMSO-d₆) δ: 156.42,156.34,153.10,152.24,148.61,147.53,142.55,134.82,130.11,126.97,125.08,124.96,117.10,116.96,115.95,113.16,112.84; HRMS (AP-ESI) calcd for C₁₈H₁₃N₄O₃ [M+H]⁺ 333.0988,found 333.0981.

4-(呋喃-2-基)-N-(4-吗啉基苯基)喹唑啉-2-胺(4x): 黄色固体,产率69%. m.p. 235 (dec.) ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 9.61 (s,1H,NH),8.57 (d,J=8 Hz,1H,ArH),8.15 (d,J=1.2 Hz,1H,ArH),7.85 (d,J=8.8 Hz,2H,ArH),7.80 (td,J=8.4,1.2 Hz,1H,ArH),7.64 (d,J=8 Hz,1H,ArH),7.52 (d,J=3.2 Hz,1H,ArH),7.39～7.35 (m,1H,ArH),6.96 (d,J=8.8 Hz,2H,ArH),6.86～6.85 (m,1H,ArH),3.76 (t,J=4.4 Hz,4H,O(CH₂)₂),3.07 (t,J =4.4 Hz,4H,N(CH₂)₂); ¹³C NMR (100 MHz,DMSO- d₆) δ: 156.92,156.33,153.63,152.30,147.09,146.64,134.49,133.66,126.87,126.71,123.89,120.49,116.74,116.35,116.12,112.99,66.67,49.78; HRMS (AP-ESI) calcd for C₂₂H₂₁N₄O₂ [M+H]⁺ 373.1665,found 373.1657.

3.2.1 中间体2及3a～3f的合成

中间体2的合成参考相关文献^{[24, 25]}.

中间体3a～3f的合成. 将化合物2 (0.20 g,1 mmol)、R¹B(OH)₂ (1 mmol)、Na₂CO₃或K₂CO₃或Cs₂CO₃ (3 mmol)、Pd(PPh₃)₄ (0.03～0.05 mmol)置于50 mL两颈瓶中,以甲苯(或二氧六环等)为溶剂,氮气(N₂)保护下于90～110 ℃反应. 用薄层色谱(TLC)监测反应,反应结束后向反应体系中加适量水并用EtOAc萃取、硫酸镁干燥、浓缩或者直接过滤然后用减压蒸馏蒸干溶剂,经柱层析得到所要产物.

2-氯-4-苯基喹唑啉(3a): 白色固体,产率78%. m.p. 108～110 ℃(文献值^[26] 112～114 ℃); ¹H NMR (300 MHz,DMSO-d₆) δ: 8.14～8.04 (m,3H,ArH),7.84～7.77 (m,3H,ArH),7.71～7.62 (m,3H,ArH).

2-氯-4-(4-甲基苯基)喹唑啉(3b): 白色固体,产率72%. m.p. 132～134 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 8.16 (d,J=8.7 Hz,1H,ArH),8.11～8.03 (m,2H,ArH),7.80～7.72 (m,3H,ArH),7.48 (d,J=8.1 Hz,2H,ArH),2.46 (s,3H,CH₃). HRMS (AP-ESI) calcd for C₁₅H₁₂ClN₂ [M+H]⁺ 255.0689,found 255.0690.

2-氯-4-(3-硝基苯基)喹唑啉(3c): 白色固体,产率53%. m.p. 190～192 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 8.61 (t,J=1.8 Hz,1H,ArH),8.53～8.50 (m,1H,ArH),8.29 (dt,J=7.8,1.2 Hz,1H,ArH),8.18～8.09 (m,3H,ArH),7.94 (t,J=8.1 Hz,1H,ArH),7.85～7.79 (m,1H,ArH). HRMS (AP-ESI) calcd for C₁₄H₉ClN₃O₂ [M+H]⁺ 286.0383,found 286.0377.

2-氯-4-(噻吩-3-基)喹唑啉(3d): 白色固体,产率34%. m.p. 136～138 ℃; ¹H NMR (300 MHz,DMSO-d₆) δ: 8.41～8.38 (m,2H,ArH),8.14～8.08 (m,1H,ArH),8.04 (m,1H,ArH),7.87～7.79 (m,2H,ArH),7.70 (dd,J=5.1,1.2 Hz,1H,ArH). HRMS (AP-ESI) calcd for C₁₂H₈Cl- N₂S [M+H]⁺ 247.0097,found 247.0092.

2-氯-4-(噻吩-2-基)喹唑啉(3e): 白色固体,产率41%. m.p. 104～106 ℃(文献值^[26] 107～108 ℃); ¹H NMR (400 MHz,DMSO-d₆) δ: 8.64 (d,J=8.0 Hz,1H,ArH),8.18～8.17 (m,1H,ArH),8.11～8.08 (m,2H,ArH),8.02 (d,J=8.0 Hz,1H,ArH),7.86～7.84 (m,1H,ArH),7.41 (dd,J=4.8,3.6 Hz,1H,ArH).

2-氯-4-(呋喃-2-基)喹唑啉(3f): 白色固体,产率34%. m.p. 143～145 ℃; ¹H NMR (400 MHz,DMSO-d₆) δ: 8.93 (d,J=8.8 Hz,1H,ArH),8.26 (s,1H),8.11 (t,J=7.2 Hz,1H,ArH),7.99 (d,J=8.0 Hz,1H,ArH),7.87 (t,J=7.2 Hz,1H,ArH),7.77 (d,J=3.6 Hz,1H,ArH),6.92 (br s,1H,ArH). HRMS (AP-ESI) calcd for C₁₂H₈ClN₂O [M+H]⁺ 231.0325,found 231.0322.

1. [1]
  Jatav, V.; Mishra, P.; Kashaw, S. Eur. J. Med. Chem. 2008, 43, 1945. doi: 10.1016/j.ejmech.2007.12.003
2. [2]
  Goyal, M.; Sasmal, D. J. Ethnopharmacol. 2014, 151, 536. doi: 10.1016/j.jep.2013.11.011
3. [3]
  Smits, R. A.; Adami, M.; Istyastono, E. P. J. Med. Chem. 2010, 53, 2390. doi: 10.1021/jm901379s
4. [4]
  Jain, K. S.; Bariwal, J. B.; Kathiravan, M. K. Bioorg. Med. Chem. 2008, 16, 4759. doi: 10.1016/j.bmc.2008.02.091
5. [5]
  Alagarsamy, V.; Pathak, U. S. Bioorg. Med. Chem. 2007, 15, 3457. doi: 10.1016/j.bmc.2007.03.007
6. [6]
  Ismail, M. A.; Barker, S.; Abou El Ella, D. A. J. Med. Chem. 2006, 49, 1526. doi: 10.1021/jm050232e
7. [7]
  Verhaeghe, P.; Azas, N.; Gasquet, M. Bioorg. Med. Chem. Lett. 2008, 18, 396. doi: 10.1016/j.bmcl.2007.10.027
8. [8]
  Pandey, S. K.; Singh, A.; Singh, A. Eur. J. Med. Chem. 2009, 44, 1188. doi: 10.1016/j.ejmech.2008.05.033
9. [9]
  Grover, G.; Kini, S. G. Eur. J. Med. Chem. 2006, 41, 256. doi: 10.1016/j.ejmech.2005.09.002
10. [10]
  Maggio, B.; Daidone, G.; Raffa, D. Eur. J. Med. Chem. 2001, 36, 737. doi: 10.1016/S0223-5234(01)01259-4
11. [11]
  Li, D.-D.; Hou, Y.-P.; Wang, W. Curr. Med. Chem. 2012, 19, 871.
12. [12]
  Khan, I.; Ibrar, A.; Abbas, N.; Saeed, A. Eur. J. Med. Chem. 2014, 76, 193. doi: 10.1016/j.ejmech.2014.02.005
13. [13]
  Connolly, D. J.; Cusack, D.; O'sullivan, T. P. Tetrahedron. 2005, 61, 10153. doi: 10.1016/j.tet.2005.07.010
14. [14]
  Hwang, D.-F.; Shiu, Y.-C.; Hwang, P.-A. J. Food Prot. 2002, 65, 1341.
15. [15]
  Hwang, D.-F.; Hsieh, Y.-W.; Shiu, Y.-C. J. Food Prot. 2002, 65, 389.
16. [16]
  Pao, W.; Miller, V.; Zakowski, M. Proc. Natl. Acad. Sci. 2004, 101, 13306. doi: 10.1073/pnas.0405220101
17. [17]
  Raymond, E.; Faivre, S.; Armand, J. P. Drugs 2000, 60, 15. doi: 10.2165/00003495-200060001-00002
18. [18]
  Wells, S. A.; Robinson, B. G.; Gagel, R. F. J. Clin. Oncol. 2012, 30, 134. doi: 10.1200/JCO.2011.35.5040
19. [19]
  Backes, A.; Zech, B.; Felber, B. Expert Opin. Drug Discovery 2008, 12, 1409.
20. [20]
  Backes, A.; Zech, B.; Felber, B. Expert Opin. Drug Discovery 2008, 12, 1427.
21. [21]
  Mclaughlin, N. P.; Evans, P. J. Org. Chem. 2009, 75, 518.
22. [22]
  Asghar, U; Witkiewicz, A. K.; Turner, N. C. Nat. Rev. Drug Discovery 2015, 14, 130. doi: 10.1038/nrd4504
23. [23]
  Wang, J. H.; Wang, Q. D.; Dun, Y. Y. Chem. J. Chin. Univ. 2014, 35, 1189 (in Chinese). (王军华, 王泉德, 顿艳艳, 高等学校化学学报, 2014, 35, 1189.)
24. [24]
  Sun, Z.; Wang, H.; Wen, K. J. Org. Chem. 2011, 76, 4149. doi: 10.1021/jo2003715
25. [25]
  Jiang, N.; Zhai, X.; Zhao, Y. Eur. J. Med. Chem. 2012, 54, 534. doi: 10.1016/j.ejmech.2012.05.039
26. [26]
  Harden, D. B. J. Org. Chem. 1988, 53(17), 4137. doi: 10.1021/jo00252a057

图式1 目标化合物的合成

Scheme 1 Synthetic route of the target compounds

下载: 全尺寸图片幻灯片

图 1 目标化合物浓度为50 μmol/L时对Hela细胞的抑制作用

Figure 1 Inhibitions of all the target compounds to Hela cell line at 50 μmol/L

下载: 全尺寸图片幻灯片

表 1 化合物4的结构

Table 1. The structure of the compounds 4

Compd.	R¹	R²
4a	Ph	H
4b	Ph	4-CH₃
4c	Ph	3-NO₂
4d	Ph	3-Morpholino
4e	Ph	4-Morpholino
4f	Ph	4-SO₂NH₂
4g	4-CH₃C₆H₄	4-CH₃
4h	4-CH₃C₆H₄	3-NO₂
4i	4-CH₃C₆H₄	4-Morpholino
4j	3-NO₂C₆H₄	H
4k	3-NO₂C₆H₄	4-CH₃
4l	3-NO₂C₆H₄	3-NO₂
4m	3-NO₂C₆H₄	4-Morpholino
4n	Thiophen-3-yl	H
4o	Thiophen-3-yl	4-CH₃
4p	Thiophen-3-yl	3-NO₂
4q	Thiophen-3-yl	4-Morpholino
4r	Thiophen-3-yl	4-CH₃
4s	Thiophen-3-yl	3-NO₂
4t	Thiophen-3-yl	4-Morpholino
4u	Furan-2-yl	H
4v	Furan-2-yl	4-CH₃
4w	Furan-2-yl	3-NO₂
4x	Furan-2-yl	4-Morpholino

下载: 导出CSV

表 2 代表化合物的体外抗增殖活性

Table 2. Antiproliferative activities of representative compounds

Compd.	IC₅₀/(μmol•L^-1)
Compd.	Hela	A549	MCF-7
4u	12.76±2.36	32.95±1.17	16.21±2.00
4v	14.93±1.59	30.47±1.56	26.24±0.76
R-Roscovitine	14.01±2.62	78.70±6.03	12.72±1.25

下载: 导出CSV

扫一扫看文章

计量

PDF下载量: 0
文章访问数: 1102
HTML全文浏览量: 160

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142