新型四氢苯并[4', 5']噻吩并[3', 2':5, 6]吡啶并[4, 3-<i>d</i>]嘧啶类化合物衍生物的合成及抗肿瘤活性

引用本文: 王胜红, 韩潇, 毛婷, 曾慧君, 陈瑞, 陈慧琼. 新型四氢苯并[4', 5']噻吩并[3', 2':5, 6]吡啶并[4, 3-d]嘧啶类化合物衍生物的合成及抗肿瘤活性[J]. 化学通报, 2019, 82(7): 623-629. shu

Citation: Wang Shenghong, Han Xiao, Mao Ting, Zeng Huijun, Chen Rui, Chen Huiqiong. Synthesis and Antitumor Activity Evaluation of Novel Tetrahydrobenzo[4', 5']thieno[3', 2': 5, 6]pyrido[4, 3-d]pyrimidine Derivatives[J]. Chemistry, 2019, 82(7): 623-629. shu

新型四氢苯并[4', 5']噻吩并[3', 2':5, 6]吡啶并[4, 3-d]嘧啶类化合物衍生物的合成及抗肿瘤活性

王胜红 ^1, ,
韩潇 ^{1,
,} ,
毛婷 ^1, ,
曾慧君 ^2, ,
陈瑞 ^3, ,
陈慧琼 ^4,

1.
长江职业学院武汉 430064
2.
武汉食品化妆品检验所武汉 430012
3.
武汉工程大学武汉 430205
4.
武汉爱民制药股份有限公司鄂州 436070

通讯作者: 韩潇 E-mail:hxiao12378@126.com

基金项目:
湖北省普通高等学校人文社会科学重点研究基地湖北技能型人才培养研究中心项目（2018JB009）和2017年度湖北省教育厅科研计划项目（B2017578）资助

摘要: 本文以环己酮为原料，通过氮杂Wittig反应合成了一系列结构新颖的取代四氢苯并噻吩并吡啶并嘧啶衍生物，并采用MTT法考察所合成目标化合物对CNE2、KB、MGC-803、MCF-7和PC3这5种肿瘤细胞的抑制活性。初步的生物活性结果表明，目标化合物对5种肿瘤细胞均有抑制活性，尤其是对胃癌MGC-803细胞展现出了更强的抑制活性。其中3-（4-氟苯基）-2-（（4-氟苯基）氨基）-5-甲基-8，9，10，11-四氢苯并[4'，5']噻吩并[3'，2'：5，6]吡啶并[4，3-d]嘧啶-4（3H）-酮[化合物8c，IC₅₀=（0.9±0.25）μmol·L^-1]对MGC-803的活性最强，是5-氟尿嘧啶[IC₅₀=（18.4±1.43）μmol·L^-1]的20倍；同时，目标化合物对正常的胃黏膜上皮细胞GES-1没有毒性。四氢苯并[4'，5']噻吩并[3'，2'：5，6]吡啶并[4，3-d]嘧啶类化合物具有良好的抗肿瘤活性，值得进一步深入研究。

关键词:

English

Synthesis and Antitumor Activity Evaluation of Novel Tetrahydrobenzo[4', 5']thieno[3', 2': 5, 6]pyrido[4, 3-d]pyrimidine Derivatives

1.
Changjiang Polytechnic, Wuhan 430064
2.
Wuhan Institute for Food and Cosmetic Control, Wuhan 430012
3.
Wuhan Institute of Technology, Wuhan 430205
4.
Wuhan Aimin Pharmaceutical Limited company, Ezhou 436070

Corresponding author: Han Xiao, hxiao12378@126.com

Received Date: 24 February 2019
Accepted Date: 16 March 2019
Available Online: 01 July 2019

Abstract: A series of novel tetrahydrobenzo[4', 5']thieno[3', 2':5, 6]pyrido[4, 3-d]pyrimidine derivatives were designed and synthesized via a tandem aza-Wittig reaction, and their anti-tumor activities in vitro were evaluated for CNE2, KB, MGC-803, MCF-7 and PC3 cells by MTT assay. The preliminary biological results showed that these conjugates display significant antiproliferative effect on these five cancer cells, especially for MGC-803. Compound 8c (IC₅₀=0.9±0.25μmol·L^-1) stood out as the most potent compound against MGC-803, and is 20 times more active than 5-FU (IC₅₀=18.4±1.43μmol·L^-1). Additionally, all compounds were nontoxic to health GES-1 and VERO cells, while 5-FU showed essential toxicity. Tetrahydrobenzo[4', 5'] thieno[3', 2':5, 6] pyrido[4, 3-d] pyrimidine compounds displayed good anti-tumor activity and deserved further research.

Key words:

杂环化合物广泛的分布在自然界中，许多重要的物质如核糖、核酸、血红素、临床疗效显著的药物，以及一些高效低毒的农药等都含杂环结构^[1~3]。因此，杂环化合物在医药和农药领域发挥着重要的作用。噻吩并嘧啶衍生物是一类具有杀菌、抗过敏、除草、抗惊厥和抗癌等良好生物活性的稠杂环化合物^[4~8]，具有较好的临床开发价值，特别是其抗癌活性引起了人们的重视并展开了相关的研究^[9~11]。近年来研究发现，吡啶并嘧啶衍生物如吡啶并[4, 3-d]嘧啶化合物也是一类具有良好抗肿瘤活性的杂环化合物^{[12, 13]}，并对多种肿瘤细胞均有较强的抑制作用。这些噻吩并嘧啶、吡啶并嘧啶类杂环化合物不仅能够抑制多种肿瘤细胞的形成、增殖，而且还可以诱导细胞的凋亡，有望成为新型的抗肿瘤药物。然而，这些杂环化合物通常由于其自身抗肿瘤作用较弱、生物利用度低和毒性较强等缺点限制了其在临床上的应用。因此，开发出抗肿瘤活性更强、毒性更低的杂环化合物具有重要的意义。基于噻吩并嘧啶、吡啶并嘧啶骨架展现出的优秀抗肿瘤活性，我们拟合成一系列同时具有噻吩、吡啶、嘧啶环的新型抗肿瘤杂环化合物。

参考噻吩并嘧啶、吡啶并嘧啶类化合物的构效关系发现，嘧啶环上的取代基对抗肿瘤活性影响甚大^[14~16]。因此，本文应用氮杂Wittig反应合成了25个嘧啶环上具有不同取代基的新型四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶衍生物6~10。为了对其构效关系进行探讨，嘧啶环上氨基(R₁)和酰胺氮原子(R₂)均用芳基和烷基(正丙基)进行了取代；同时为了进一步研究R₁、R₂苯环上电子效应对活性的影响，给电子基(OMe)、吸电子基(F，Cl)类化合物也被合成出来。这些目标杂环化合物的具体合成路线见图式 1。首先以环己酮为原料，利用Gewald反应制得中间体2-氨基-3-氰基-4, 5, 6, 7-四氢苯并^{[4, 5]}噻吩(化合物2)^[17]；接着用无水四氯化锡为催化剂，将中间体2与乙酰乙酸乙酯缩合，制得化合物3^[4]；应用氮杂Wittig反应将膦亚胺中间体4与异氰酸酯反应，得到化合物5；最后，以无水乙腈作溶剂，在催化量乙醇钠的存在下，将化合物5与相应的胺关环得到目标化合物6~10(图式 1)。随后，对目标化合物6~10进行了初步的抗肿瘤活性筛选。

图式 1

图式 1. 四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶化合物6~10的合成路线

Scheme 1. Synthetic route of tetrahydrobenzo[4′, 5′]thieno[3′, 2′:5, 6]pyrido[4, 3-d]pyrimidine compounds 6~10

下载: 全尺寸图片幻灯片

1. 实验部分

1.1 仪器与试剂

NE×US 470型傅立叶变换红外光谱仪(美国Nicolet公司)；BrukerAM-400Hz型核磁共振谱仪(TMS内标，德国Bruker公司)；XR4显微熔点测定仪(上海光学仪器厂)；ZF-I三用紫外分析仪(上海顾村电光仪器厂)；超净工作台(北京半导体设备一厂)；BB16/BB5060 CO₂培养箱(上海力创科学仪器有限公司)；CKX31型倒置显微镜(奥林巴斯公司)；ELx800通用酶标仪(美国BioTek公司)。

所有原料、试剂和溶剂如无说明均为市售分析纯级。无水甲苯用金属钠干燥后减压蒸馏制得，无水乙腈用氢化钙干燥后减压蒸馏制得。粒径48~75 μm柱色谱硅胶(青岛海洋化工厂)；GF254硅胶薄层板(青岛海洋化工厂)。

1.2 8, 9, 10, 11-四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶-4(3H)-酮(6~10)的合成

中间体2和3分别按照文献[17]和^[4]报道的方法合成。氩气保护下，将按照文献[4]报道方法制得的中间体4(2.530g，4.6mmol)和相应的异氰酸酯(5.52mmol)依次加入到二氯甲烷(15mL)中，加热回流反应12h，减压除去溶剂，得到化合物5，不经处理直接投入下一步反应。将所得化合物5溶解到乙腈(15mL)中，加入各种不同的取代胺(4.6mmol)以及催化量的乙醇钠(20mg，0.3mmol)，加热回流反应6h，过滤，减压除去溶剂，经柱色谱纯化(淋洗剂为石油醚/二氯甲烷)得四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶-4(3H)-酮(6~10)，其理化和波谱数据见表 1和表 2。

表 1

表 1 化合物6~10的理化数据及高分辨质谱数据

Table 1. Physical contants and HR-MS of synthesized compounds 6~10

下载: 导出CSV

Compd.	R₁	R₂	Yield/%	mp/℃	Physical property	IR (KBr) /cm^-1	HR-MS (ESI)m/z ([M+H]⁺)
Compd.	R₁	R₂	Yield/%	mp/℃	Physical property	IR (KBr) /cm^-1		Calcd.	Found
6a	Ph	Ph	64.2	224~226	White solid	1671 (C=O)	C₂₆H₂₃N₄OS	439.5520	439.5525
6b	Ph	4-OMe-Ph	75.4	218~220	White solid	1678 (C=O)	C₂₇H₂₅N₄O₂S	469.5780	469.5786
6c	Ph	4-F-Ph	64.2	224~226	White solid	1671 (C=O)	C₂₆H₂₂FN₄OS	457.5425	457.5431
6d	Ph	4-Cl-Ph	61.8	234~237	White solid	1684 (C=O)	C₂₆H₂₂ClN₄OS	473.9971	473.9976
6e	Ph	C₃H₇	78.4	215~218	White solid	1672 (C=O)	C₂₃H₂₅N₄OS	405.1749	405.1753
7a	4-OMe-Ph	Ph	68.9	232~234	Yellow solid	1675 (C=O)	C₂₇H₂₅N₄O₂S	469.5780	469.5786
7b	4-OMe-Ph	4-OMe-Ph	72.1	234~237	Yellow solid	1668 (C=O)	C₂₈H₂₇N₄O₃S	499.6040	499.6044
7c	4-OMe-Ph	4-F-Ph	63.6	225~228	White solid	1675 (C=O)	C₂₇H₂₄FN₄O₂S	487.5685	487.5691
7d	4-OMe-Ph	4-Cl-Ph	74.2	244~246	White solid	1669 (C=O)	C₂₇H₂₄ClN₄O₂S	504.0231	504.0236
7e	4-OMe-Ph	C₃H₇	82.5	221~223	White solid	1676 (C=O)	C₂₄H₂₇N₄O₂S	435.5618	435.5622
8a	4-F-Ph	Ph	56.8	226~229	White solid	1672 (C=O)	C₂₆H₂₂FN₄OS	457.5425	457.5431
8b	4-F-Ph	4-OMe-Ph	63.9	223~225	White solid	1670 (C=O)	C₂₇H₂₄FN₄O₂S	487.5685	487.5691
8c	4-F-Ph	4-F-Ph	55.4	226~229	Yellow solid	1672 (C=O)	C₂₆H₂₁F₂N₄OS	475.5329	475.5333
8d	4-F-Ph	4-Cl-Ph	53.2	235~238	Yellow solid	1675 (C=O)	C₂₆H₂₁ClFN₄OS	491.9875	491.9882
8e	4-F-Ph	C₃H₇	53.2	222~225	White solid	1669 (C=O)	C₂₃H₂₄FN₄OS	423.5263	423.5267
9a	4-Cl-Ph	Ph	68.4	242~244	White solid	1678 (C=O)	C₂₆H₂₂ClN₄OS	473.9971	473.9976
9b	4-Cl-Ph	4-OMe-Ph	72.5	243~246	White solid	1671 (C=O)	C₂₇H₂₄ClN₄O₂S	504.0231	504.0236
9c	4-Cl-Ph	4-F-Ph	70.4	240~242	White solid	1677 (C=O)	C₂₆H₂₁ClFN₄OS	491.9875	491.9882
9d	4-Cl-Ph	4-Cl-Ph	67.3	255~257	White solid	1678 (C=O)	C₂₆H₂₁Cl₂N₄OS	508.4421	508.4426
9e	4-Cl-Ph	C₃H₇	61.9	246~249	Yellow solid	1672 (C=O)	C₂₃H₂₄ClN₄OS	439.9809	439.9816
10a	C₃H₇	Ph	57.8	211~213	Yellow solid	1675 (C=O)	C₂₃H₂₅N₄OS	405.1749	405.1753
10b	C₃H₇	4-OMe-Ph	54.6	214~217	Yellow solid	1665 (C=O)	C₂₄H₂₇N₄O₂S	435.5618	435.5622
10c	C₃H₇	4-F-Ph	60.7	212~215	Yellow solid	1668 (C=O)	C₂₃H₂₄FN₄OS	423.5263	423.5267
10d	C₃H₇	4-Cl-Ph	52.9	226~229	Yellow solid	1677 (C=O)	C₂₃H₂₄ClN₄OS	439.9809	439.9816
10e	C₃H₇	C₃H₇	63.7	211~214	Yellow solid	1664 (C=O)	C₂₀H₂₇N₄OS	371.5196	371.5201

表 2

表 2 化合物6~10的¹H NMR和¹³C NMR数据

Table 2. The ¹H NMR and ¹³C NMR data of compounds 6~10

下载: 导出CSV

Compd.	¹H NMR (400MHz, CDCl₃) δ	¹³C NMR (101MHz, CDCl₃) δ
6a	7.76 (d, J=8.0Hz, 2H), 7.65 (m, 3H), 7.48 (d, J=8.4Hz, 2H), 7.37 (d, J=8.0Hz, 2H), 7.11 (m, 1H), 3.19 (m, 2H), 3.09 (s, 3H), 2.88 (m, 2H), 1.91 (m, 2H)	164.0, 161.1, 157.5, 151.2, 150.1, 137.8, 136.2, 133.6, 131.2, 130.5, 129.4, 128.2, 127.9, 125.6, 124.4, 122.7, 118.4, 27.2, 26.9, 25.7, 23.1, 22.5
6b	7.79 (d, J=8.0Hz, 2H), 7.41 (d, J=8.0Hz, 2H), 7.12 (d, J=8.0Hz, 2H), 7.07 (m, 1H), 6.97 (d, J=8.4Hz, 2H), 3.79 (s, 3H, OCH₃), 3.21 (m, 2H, CH₂), 3.09 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 2H)	163.7, 163.5, 161.2, 157.4, 152.1, 150.5, 136.9, 133.6, 132.4, 131.1, 129.6, 128.5, 125.7, 124.4, 120.7, 118.8, 116.6, 55.6, 27.2, 26.9, 25.9, 22.7, 22.3
6c	7.81 (d, J=8.8Hz, 2H), 7.83 (d, J=8.4Hz, 2H), 7.52 (d, J=8.0Hz, 2H), 7.29 (d, J=8.4Hz, 2H), 7.14 (m, 1H), 3.21 (m, 2H, CH₂), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.88 (m, 4H)	165.2(d, ¹J_CF =250.4Hz), 163.7, 161.4, 157.9, 151.6, 150.2, 137.9, 136.7, 133.4, 132.2, 130.6 (d, ³J_CF =8.8Hz), 129.7, 128.1(d, ⁴J_CF =4.0Hz), 125.8, 124.4, 116.7 (d, ²J_CF =22.4Hz), 113.1, 27.1, 26.7, 25.8, 22.9, 22.6
6d	7.78 (d, J=8.4Hz, 2H), 7.62 (d, J=8.0Hz, 2H), 7.41 (d, J=8.4Hz, 2H), 7.38 (d, J=8.4Hz, 2H), 7.14 (m, 1H), 3.18 (m, 2H), 3.09 (s, 3H, CH₃), 2.87 (m, 2H), 1.91 (m, 4H)	63.9, 161.3, 158.2, 151.8, 151.1, 137.5, 136.1, 133.2, 132.1, 130.8, 130.1, 128.4, 127.5, 125.8, 124.9, 124.2, 113.1, 27.3, 26.9, 25.6, 22.7, 22.8
6e	7.80 (d, J=8.0Hz, 2H), 7.42 (d, J=8.0Hz, 2H), 7.13 (m, 1H), 3.91 (t, J=7.2Hz, 2H, CH₂), 3.19 (m, 2H), 3.10 (s, 3H, CH₃), 2.88 (m, 2H), 1.88 (m, 4H, CH₂), 1.72 (m, 2H), 0.99 (t, J=6.4Hz, 3H, CH₃)	163.2, 161.2, 157.9, 151.2, 149.5, 136.4, 134.2, 131.6, 128.7, 125.8, 124.4, 122.6, 112.7, 48.8, 27.2, 26.6, 26.5, 22.7, 22.5, 22.1, 11.3
7a	7.74 (d, J=8.0Hz, 2H), 7.61 (d, J=8.4Hz, 2H), 7.41 (d, J=7.6Hz, 2H), 7.15 (m, 1H), 6.98 (d, J=8.0Hz, 2H), 3.78 (s, 3H, OCH₃), 3.21 (m, 2H), 3.11 (s, 3H, CH₃), 2.87 (m, 2H), 1.90 (m, 4H)	163.6, 163.2, 161.4, 158.3, 151.8, 149.7, 137.6, 133.3, 132.5, 130.8, 129.8, 128.4, 127.6, 125.8, 124.9, 124.1, 118.5, 55.7, 27.7, 26.9, 25.7, 22.9, 22.5
7b	7.76 (d, J=8.4Hz, 2H), 7.71 (d, J=8.0Hz, 2H), 7.05 (d, J=7.6Hz, 2H), 6.98 (d, J=8.0Hz, 2H), 3.81 (s, 3H, OCH₃), 3.78 (s, 3H, OCH₃), 3.21 (m, 2H), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 4H)	163.8, 163.6, 163.2, 161.1, 157.9, 151.8, 149.5, 137.8, 136.6, 133.5, 132.4, 130.8, 128.7, 125.3, 124.4, 122.7, 118.8, 55.7, 55.6, 27.9, 26.8, 25.7, 22.9, 22.6
7c	7.80 (d, J=8.4Hz, 2H), 7.72 (d, J=8.0Hz, 2H), 7.21 (d, J=8.0Hz, 2H), 7.02 (d, J=8.0Hz, 2H), 3.79 (s, 3H, OMe), 3.22 (m, CH₂, 2H), 3.11 (s, 3H, CH₃), 2.89 (m, 2H), 1.89 (m, 4H)	165.1(d, ¹J_CF =254.2Hz), 163.5, 163.2, 161.0, 157.9, 151.8, 150.2, 137.9, 136.7, 133.8, 132.4, 130.7 (d, ³J_CF =7.6Hz), 129.4 (d, ⁴J_CF =3.6Hz), 125.7, 124.5, 116.6 (d, ²J_CF =22.8Hz), 113.6, 55.7, 27.9, 26.7, 25.6, 22.9, 22.7
7d	7.78 (d, J=8.0Hz, 2H), 7.65 (d, J=8.4Hz, 2H), 7.39 (d, J=7.2Hz, 2H), 7.01 (d, J=7.6Hz, 2H), 3.79 (s, 3H, OCH₃), 3.20 (m, 2H), 3.12 (s, 3H, CH₃), 2.87 (m, 2H), 1.91 (m, 4H)	163.5, 163.2, 161.2, 159.6, 157.8, 150.3, 148.9, 138.4, 137.4, 132.6, 130.7, 129.6, 129.3, 128.6, 126.3, 124.4, 122.5, 55.9, 27.7, 26.9, 26.5, 22.9, 22.6
7e	7.72 (d, J=8.0Hz, 2H), 6.97 (d, J=7.2Hz, 2H), 3.96 (t, J=6.4Hz, 2H), 3.20 (m, 2H), 3.10 (s, 3H, CH₃), 3.78 (s, 3H, OCH₃), 2.88 (m, 2H), 1.90 (m, 4H), 1.79 (m, 2H), 1.01 (t, J=7.2Hz, 3H, CH₃)	163.3, 161.2, 160.2, 157.9, 151.4, 149.8, 133.5, 131.4, 129.7, 127.1, 125.3, 124.4, 120.5, 55.8, 48.8, 27.9, 27.6, 26.7, 22.9, 22.6, 22.5, 11.3
8a	7.85 (d, J=8.4Hz, 2H), 7.76 (d, J=8.0Hz, 2H), 7.54 (d, J=8.0Hz, 2H), 7.17 (d, J=8.0Hz, 2H), 7.15 (m, 1H), 3.20 (m, 2H, CH₂), 3.10 (s, 3H, CH₃), 2.87 (m, 2H), 1.90 (m, 4H)	165.3(d, ¹J_CF =262.8Hz), 163.4, 161.1, 157.6, 151.2, 149.6, 137.7, 136.5, 133.2, 132.1, 130.7 (d, ³J_CF =9.2Hz), 129.5, 128.4 (d, ⁴J_CF =4.4Hz), 125.9, 124.5, 116.6 (d, ²J_CF =24.0Hz), 113.4, 27.2, 26.9, 25.7, 22.7, 22.5
8b	7.84 (d, J=8.0Hz, 2H), 7.41 (d, J=8.4Hz, 2H), 7.15 (d, J=8.4Hz, 2H), 6.94 (d, J=8.4Hz, 2H), 3.78 (s, 3H, OMe), 3.21 (m, 2H, CH₂), 3.11 (s, 3H, CH₃), 2.88 (m, 2H), 1.91 (m, 2H)	165.3(d, ¹J_CF =261.6Hz), 163.1, 161.2, 157.7, 151.8, 149.6, 137.7, 136.4, 133.7, 132.6, 130.9 (d, ³J_CF =8.4Hz), 129.1, 128.7 (d, ⁴J_CF =4.0Hz), 125.5, 124.4, 111.7 (d, ²J_CF =23.6Hz), 113.6, 55.8, 27.8, 26.9, 25.5, 22.9, 22.6
8c	7.84 (d, J=8.4Hz, 2H), 7.76 (d, J=8.8Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 7.16 (d, J=8.0Hz, 2H), 3.20 (m, 2H, CH₂), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 4H)	165.3(d, ¹J_CF =252.4Hz), 165.1 (d, ¹J_CF =254.8Hz), 163.4, 161.0, 157.5, 151.7, 150.1, 137.9, 132.2, 131.1 (d, ³J_CF =7.2Hz), 130.8 (d, ³J_CF =8.4Hz), 129.4 (d, ⁴J_CF =3.2Hz), 128.6 (d, ⁴J_CF =3.6Hz), 125.5, 117.4 (d, ²J_CF =24.4Hz), 116.6 (d, ²J_CF =23.6Hz), 113.2, 27.5, 26.9, 25.5, 22.8, 22.5
8d	7.86 (d, J=8.0Hz, 2H), 7.82 (d, J=8.8Hz, 2H), 7.18 (d, J=8.4Hz, 2H), 7.15 (d, J=8.0Hz, 2H), 3.21 (m, 2H, CH₂), 3.09 (s, 2H, CH₃), 2.88 (m, 2H), 1.91 (m, 2H)	165.4, 165.2(d, ¹J_CF =254.6Hz), 163.2, 161.1, 157.7, 151.8, 149.5, 137.6, 136.8, 133.2, 132.4, 130.5 (d, ³J_CF =8.4Hz), 129.6 (d, ⁴J_CF =4.0Hz), 125.6, 124.4, 116.9 (d, ²J_CF =23.2Hz), 113.3, 27.5, 26.9, 25.7, 22.9, 22.6
8e	7.85 (d, J=8.0Hz, 2H), 7.16 (d, J=8.0Hz, 2H), 3.93 (t, J=7.2Hz, 2H), 3.20 (m, 2H), 3.09 (s, 3H, CH₃), 2.87 (m, 2H), 1.91 (4H, m), 1.79 (m, 2H), 1.01 (t, J=6.8Hz, 3H, CH₃)	165.2(d, ¹J_CF =236.4Hz), 163.4, 161.3, 157.9, 151.7, 150.1, 137.8, 130.7 (d, ³J_CF =8.0Hz), 129.9, 128.3 (d, ⁴J_CF =4.0Hz), 124.6, 116.5 (d, ²J_CF =22.8Hz), 113.4, 48.8, 27.2, 26.8, 25.9, 22.9, 22.7, 22.5, 11.3
9a	7.80 (d, J=8.4Hz, 2H), 7.68 (d, J=8.0Hz, 2H), 7.54 (d, J=8.0Hz, 2H), 7.46 (d, J=8.4Hz, 2H), 7.15 (m, 1H), 3.22 (m, 2H), 3.12 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 4H)	163.4, 161.2, 159.4, 157.7, 150.1, 148.8, 138.5, 136.7, 133.4, 131.6, 130.4, 129.8, 129.3, 126.1, 124.2, 122.4, 121.5, 27.2, 26.9, 26.1, 22.9, 22.8
9b	7.81 (d, J=8.8Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 7.11 (d, J=7.6Hz, 2H), 7.04 (d, J=7.6Hz, 2H), 3.78 (s, 3H, OCH₃), 3.21 (m, 2H), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.89 (m, 4H)	163.6, 163.2, 161.2, 159.9, 157.4, 149.6, 148.5, 138.4, 137.2, 132.6, 130.7, 129.8, 129.3, 128.6, 126.0, 124.5, 122.6, 55.9, 27.2, 26.9, 26.3, 22.9, 22.7
9c	7.82 (d, J=8.0Hz, 2H), 7.80 (d, J=8.4Hz, 2H), 7.45 (d, J=8.4Hz, 2H), 7.15 (d, J=8.4Hz, 2H), 3.21 (m, CH₂, 2H), 3.12 (s, 3H, CH₃), 2.88 (m, 2H), 1.91 (m, 4H)	65.4(d, ¹J_CF =225.2Hz), 161.6, 159.7, 157.8, 150.1, 148.9, 138.5, 137.1, 132.3, 130.8 (d, ³J_CF =7.6Hz), 129.5, 129.3, 128.4 (d, ⁴J_CF =4.4Hz), 126.0, 124.6, 116.9 (d, ²J_CF =23.2Hz), 113.6, 27.1, 26.9, 26.5, 22.9, 22.6
9d	7.83 (d, J=8.0Hz, 2H), 7.78 (d, J=8.0Hz, 2H), 7.51 (d, J=7.6Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 3.22 (m, 2H), 3.11 (s, 3H, CH₃), 2.89 (m, 2H), 1.92 (m, 4H)	163.6, 161.0, 159.6, 157.8, 150.1, 148.9, 138.7, 136.9, 133.2, 131.8, 130.2, 129.9, 129.6, 126.5, 124.7, 123.8, 122.5, 27.1, 26.7, 25.8, 22.8, 22.6
9e	7.81 (d, J=8.0Hz, 2H), 7.42 (d, J=8.4Hz, 2H), 3.93 (t, J=7.2Hz, 2H), 3.23 (m, 2H), 3.12 (s, 3H, CH₃), 2.89 (m, 2H), 1.91 (m, 4H), 1.78 (m, 2H), 1.01 (t, J=7.2Hz, 3H)	163.4, 161.3, 159.7, 157.6, 149.7, 148.3, 137.6, 133.8, 130.2, 124.6, 122.4, 118.4, 112.8, 48.9, 27.2, 26.9, 26.1, 22.9, 22.8, 22.5, 11.3
10a	7.86 (d, J=8.4Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 7.12 (m, 1H), 3.94 (t, J=7.6Hz, 2H), 3.20 (m, 2H), 3.10 (s, 3H, CH₃), 2.88 (m, 2H), 1.91 (m, 4H), 1.78 (m, 2H), 0.98 (t, J=7.2Hz, 3H, CH₃)	163.0, 160.5, 157.8, 151.1, 149.4, 136.2, 133.8, 131.1, 128.5, 125.6, 124.7, 122.3, 112.5, 48.4, 27.1, 26.9, 26.1, 22.6, 22.3, 22.1, 11.4
10b	7.73 (d, J=8.4Hz, 2H), 6.96 (d, J=7.6Hz, 2H), 3.97 (t, J=6.8Hz, 2H), 3.18 (m, 2H), 3.11 (s, 3H, CH₃), 3.78 (s, 3H, OCH₃), 2.86 (m, 2H), 1.91 (m, 4H), 1.79 (m, 2H), 1.00 (t, J=7.2Hz, 3H, CH₃)	163.0, 161.4, 160.2, 157.8, 151.4, 150.7, 133.2, 131.4, 129.6, 128.2, 125.1, 123.4, 119.2, 55.8, 48.6, 27.6, 27.1, 26.2, 23.8, 22.6, 22.0, 11.5
10c	7.28 (d, J=8.8Hz, 2H), 7.07 (d, J=8.0Hz, 2H), 3.93 (t, J=7.6Hz, 2H), 3.20 (m, 2H), 3.09 (s, 3H, CH₃), 2.86 (m, 2H), 1.89 (m, 4H), 1.74 (m, 2H), 1.02 (t, J=7.2Hz, 3H, CH₃)	165.4(d, ¹J_CF =228.8Hz), 163.2, 161.2, 159.4, 157.6, 150.4, 149.6, 137.4, 130.2 (d, ³J_CF =8.0Hz), 128.6 (d, ⁴J_CF =3.2Hz), 120.4, 117.8 (d, ²J_CF =22.4Hz), 112.3, 49.1, 27.5, 26.7, 26.1, 23.4, 22.8, 22.6, 11.3
10d	7.47 (d, J=8.0Hz, 2H), 7.23 (d, J=7.6Hz, 2H), 3.94 (t, J=6.4Hz, 2H), 3.17 (m, 2H), 3.09 (s, 3H, CH₃), 2.89 (m, 2H), 1.92 (m, 4H), 1.77 (m, 2H), 0.88 (t, J=7.2Hz, 3H, CH₃)	163.1, 160.8, 157.2, 151.1, 150.2, 136.7, 133.7, 130.0, 128.9, 126.7, 125.6, 122.3, 113.2, 49.4, 27.8, 27.3, 25.9, 23.2, 22.9, 22.5, 11.4
10e	3.95 (t, J=7.2Hz, 2H), 3.21 (m, 2H), 3.21 (m, 2H), 3.09 (s, 3H, CH₃), 2.88 (m, 2H), 1.90 (m, 4H), 1.78 (m, 2H), 1.71 (m, 2H), 1.22 (t, J=7.2Hz, 3H, CH₃), 1.01 (t, J=6.8Hz, 3H, CH₃)	163.2, 161.1, 157.4, 150.8, 149.5, 137.2, 128.3, 125.1, 120.8, 48.9, 47.6, 27.1, 26.9, 25.9, 23.1, 22.9, 22.7, 22.5, 11.6, 11.4

1.3 抗肿瘤活性实验

选取鼻咽鳞癌CNE2、口腔表皮样癌KB、胃癌MGC-803、乳腺癌MCF-7和前列腺癌PC3细胞为测试细胞株，正常的胃黏膜细胞GES-1为对照细胞株，以5-氟尿嘧啶(5-FU)为阳性对照药，采用MTT法对合成的化合物进行抗肿瘤活性评价。取对数生长期的测试细胞株接种于96孔细胞培养板中，待细胞完全贴壁后，弃去原培养液，加入100μL的含有测试药物的培养液药物培养72h后，每孔加入5g·L^-1 MTT (30μL)，置于37℃、体积分数5% CO₂培养箱中继续孵育4h，弃去培养液，然后每孔加入100μL DMSO溶解。使用酶标仪在570nm波长测定每孔的吸光度(OD)值，分析实验结果，并计算出IC₅₀，结果见表 3。

表 3

表 3 化合物6~10的抗肿瘤活性结果(n=3)

Table 3. Whole cell antiproliferative activity of compounds 6~10 (n=3)

下载: 导出CSV

Compd.	IC₅₀/(μmol·L^-1)
Compd.	CNE2	KB	MGC-803	MCF-7	PC3
6a	9.7±0.87	10.5±1.03	9.7±0.56	20.1±1.39	17.8±1.44
6b	12.1±1.66	9.9±0.89	8.5±0.42	23.4±1.61	12.6±1.52
6c	8.1±0.74	9.1±0.57	6.4±0.29	18.2±1.72	11.9±0.96
6d	9.7±0.79	11.9±1.31	11.7±1.55	21.8±2.45	20.2±1.73
6e	11.4±1.59	12.1±1.63	14.8±1.92	17.7±1.17	23.5±2.46
7a	11.7±1.85	14.4±2.41	11.5±1.67	24.2±2.32	23.7±2.55
7b	9.6±2.38	12.7±1.49	10.8±2.23	19.1±1.57	20.2±2.78
7c	11.2±1.81	9.9±1.31	8.7±0.78	22.5±1.99	15.5±1.19
7d	13.4±1.46	14.8±1.48	11.6±1.11	11.5±1.22	17.8±2.11
7e	14.8±1.09	15.3±1.76	13.3±1.40	20.8±2.08	13.3±1.71
8a	9.4±0.34	8.6±0.52	7.2±0.66	14.6±1.16	22.5±2.11
8b	8.3±0.77	4.2±0.61	2.4±0.72	17.5±1.63	16.7 ±1.56
8c	3.3±0.54	2.1±0.27	0.9±0.25	9.1±0.99	13.7±1.44
8d	7.9±0.52	3.9±0.50	5.5±0.16	10.6±1.35	17.3±1.27
8e	13.7±0.87	10.5±1.18	9.7±0.93	21.8±2.67	22.5±1.89
9a	14.3±1.17	12.8±1.15	7.8±0.94	17.2±2.27	15.9±1.65
9b	12.4±1.93	9.9±0.58	8.6±0.83	14.4±1.10	13.4±1.33
9c	7.8±0.56	6.4±0.76	4.2±0.45	8.9±0.76	16.3±2.77
9d	9.3±0.82	7.8±0.69	6.9±0.42	13.8±1.52	11.4±1.82
9e	11.5±1.79	10.1±1.76	11.7±2.11	21.7±2.43	17.3±1.35
10a	18.4±2.25	16.5±1.43	16.4±1.73	18.7±2.14	16.6±1.71
10b	21.7±1.84	25.3±2. 74	18.9±1.04	16.4±1.57	21.3±2.18
10c	14.4±1.01	11.5±1.62	14.8±1.14	13.8±1.79	18.2±1.64
10d	11.9±1.47	17.8±1.13	15.1±1.29	18.1±1.35	17.7±1.51
10e	25.2±1.51	22.3±2.14	19.3±2.55	15.8±1.29	21.6±2.26
5-FU	17.8±1.36	12.5±0.87	18.4±1.43	10.8±0.96	15.4±1.22

2. 结果与讨论

2.1 合成部分

本实验合成四氢苯并噻酚并吡啶并嘧啶类化合物6~10的关键步骤是中间体2和3的制备。目前合成2-氨基噻吩衍生物2最简便有效的方法是Gewald反应^[17]。在制备中间体3过程中，四氯化锡在反应中起着非常重要的作用，它既是脱水剂也是催化剂^[18]。然而，无水四氯化锡对湿敏感，易水解析出α-锡酸，此外四氯化锡具有刺激性、腐蚀性和中等毒。因此，在反应中需采用气体保护来减少反应装置里的水和毒性无水四氯化锡的流失，同时通过加入过量的无水四氯化锡来提高产率。

2.2 抗肿瘤活性

体外抗增殖活性的结果(表 3)表明，这些新型的四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶衍生物对CNE2、KB和MGC-803有较强的抑制活性，而对MCF-7和PC3仅展现出了中等的抑制活性；此外，嘧啶环上R₁和R₂取代基对活性影响较大。总的来说，R₁和R₂为F取代的化合物对CNE2、KB和MGC-803的抑制活性要强于其他取代基类化合物。例如在化合物6(R₁=Ph)系列中，R₂为4-氟苯基取代的化合物6c对CNE2、MGC-803和MCF-7肿瘤细胞株的抑制活性较强于同系列的其他化合物6a、6b、6d和6e；进一步在R₁的苯环上引入给电子基(OMe，化合物7a~7e)也没有显著地增加抗肿瘤活性，部分化合物甚至降低了抗癌活性，如化合物7c[IC₅₀=(8.7±0.78)μmol·L^-1]对胃癌MGC-803细胞的抑制活性较化合物6c[IC₅₀=(6.4±0.29)μmol·L^-1]降低了1.4倍；然而，在R₁的苯环上引入吸电子基F和Cl(化合物8a~8e和9a~9e)却能显著增加抗肿瘤活性，尤其是R₁和R₂均为4-氟苯基取代的化合物8c对CNE2、KB和MGC-803这三种肿瘤细胞的抑制活性较6c显著地提高；但是将R₁用烷基取代后(化合物10a~10e)，抗肿瘤活性却明显降低，其中是R₁和R₂均为烷基取代的化合物10e对5种肿瘤细胞的抑制活性均较弱(IC₅₀>20μmol·L^-1)。这些结果说明，在四氢苯并噻吩并吡啶并嘧啶骨架的嘧啶环上(R₁或R₂)引入烷基将降低化合物的抗肿瘤活性，而4-氟苯基能够增加抗肿瘤活性，尤其是R₁和R₂均为4-氟苯基取代的化合物8c对CNE2、KB和MGC-803这3种肿瘤细胞的抑制活性最强，甚至是阳性药物5-FU的4倍以上。

另外，为了进一步考察这类新型四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶衍生物的毒性，用正常的胃黏膜细胞GES-1作为对照来检测对胃癌MGC-803细胞有较强抑制活性化合物6a~6c、7c、8a~8e、9a~9d(IC₅₀<10μmol·L^-1)的毒性，并计算出其体外治疗指数(In vitro therapeutic index，IVTI)，化合物的IVTI值越高，安全性越高^[19]。如表 4所示，这些新型的氮杂环化合物对正常的胃黏膜细胞GES-1和非洲绿猴肾细胞VERO没有毒性，而阳性药物5-FU却展现出了一定的毒性；同时，化合物6a~6c、7c、8a~8e、9a~9d的ITVI也高于5-FU。

表 4

表 4 化合物6a~6c、7c、8a~8e、9a~9d和5-FU的体外治疗指数(IVTI)

Table 4. In vitro therapeutic index of compounds 6a~6c, 7c, 8a~8e, 9a~9d and 5-FU

下载: 导出CSV

Compd.	MGC-803	GES-1	IVTI
6a	9.7±0.56	>100	>10.31
6b	8.5±0.42	>100	>11.76
6c	6.4±0.29	>100	>15.62
7c	8.7±0.78	>100	>11.49
8a	7.2±0.66	>100	>13.88
8b	2.4±0.72	>100	>41.66
8c	0.9±0.25	>100	>111.11
8d	5.5±0.16	>100	>18.18
8e	9.7±0.93	>100	>10.30
9a	7.8±0.94	>100	>12.82
9b	8.6±0.83	>100	>11.62
9c	4.2±0.45	>100	>23.80
9d	6.9±0.42	>100	>14.49
5-FU	18.4±1.43	10.7±0.59	0.58
IVTI-The ratio of IC₅₀(GES-1) and IC₅₀(MGC-803)

3. 结论

本文通过氮杂Wittig反应合成了一系列新型的四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶类化合物。经过测试其抗瘤活性发现，这类新型的氮杂环化合物对CNE2、KB、MGC-803、MCF-7和PC3肿瘤细胞均有抑制活性，其中嘧啶环上R₁和R₂均为4-氟苯基的化合物8c对肿瘤细胞CNE2、KB和MGC-803展现出了最强的抑制活性，并强于阳性药物5-FU；同时，这类氮杂化合物对正常GES-1细胞没有毒性。这些结果说明四氢苯并[4′, 5′]噻酚并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶骨架是一个具有抗肿瘤活性的药效团，值得更进一步研究。

1. [1]
  丁明武. 中国科学:化学, 2010, 40(7):869-877. http://www.cnki.com.cn/Article/CJFDTotal-JBXK201007007.htm
2. [2]
  赵培亮, 游文玮, 段安娜. 药学学报, 2012, 47(5):580-587. http://www.cnki.com.cn/Article/CJFDTotal-YXXB201205006.htm
3. [3]
  尚慧杰, 闫强, 吴书敏等. 沈阳药科大学学报, 2015, 32(8):604-607. http://www.cnki.com.cn/Article/CJFDTotal-SYYD201508005.htm
4. [4]
  任青云, 贺红武, 刘建超. 有机化学, 2007, 27(3):397-401. http://www.cnki.com.cn/Article/CJFDTotal-YJHU200703012.htm
5. [5]
  曾国平, 郑平. 化学学报, 2012, 70(6):759-764. http://www.cnki.com.cn/Article/CJFDTotal-HXXB201206011.htm
6. [6]
  姜楠, 李婷, 陶海燕等. 沈阳药科大学学报, 2012, 29(11):829-833. http://www.cnki.com.cn/Article/CJFDTotal-SYYD201211002.htm
7. [7]
  C H Wu, M S Coumar, C Y Chu et al. J. Med. Chem., 2010, 53(20):7316-7326. doi: 10.1021/jm100607r
8. [8]
  陈玉, 柏舜, 贺红武等. 有机化学, 2014, 34(5):2362-2369. http://www.cnki.com.cn/Article/CJFDTotal-YJHU201411024.htm
9. [9]
  Q Y Ren, Y J Liang, H W He et al. Bioorg. Med. Chem. Lett., 2009, 19(23):6713-6716. doi: 10.1016/j.bmcl.2009.09.117
10. [10]
  张广龙, 欧阳贵平, 李文举等. 精细化工中间体, 2010, 40(3):12-17. http://www.cnki.com.cn/Article/CJFDTotal-HNHG201003007.htm
11. [11]
  A T Mavrova, S Dimov, D Yancheva et al. Eur. J. Med. Chem., 2016, 123:69-79. doi: 10.1016/j.ejmech.2016.07.022
12. [12]
  H Abe, S Kikuchi, K Hayakawa et al. ACS Med. Chem. Lett., 2011, 2(4):320-324. doi: 10.1021/ml200004g
13. [13]
  W Zhou, X F Liu, Z C Tu et al. J. Med. Chem., 2013, 56(20):7821-7837. doi: 10.1021/jm401045n
14. [14]
  J Lou, Z Liu, Y Li et al. Bioorg. Med. Chem. Lett., 2011, 21(22):6662-6666. doi: 10.1016/j.bmcl.2011.09.061
15. [15]
  Y Loidreau, P Marchand, C Dubouilh-Benard et al. Eur. J. Med. Chem., 2012, 58:171-183. doi: 10.1016/j.ejmech.2012.10.006
16. [16]
  W Y Mo, Y J Liang, Y C Gu et al. Bioorg. Med. Chem. Lett., 2011, 21(19):5975-5977. doi: 10.1016/j.bmcl.2011.07.067
17. [17]
  L Aurelio, C Valant, H Figler et al. Bioorg. Med. Chem., 2009, 17(20):7353-7361. doi: 10.1016/j.bmc.2009.08.024
18. [18]
  Y Ai, Y J Liang, J C Liu et al. Eur. J. Med. Chem., 2012, 47(1):206-213.
19. [19]
  P K Muller, M N Molton. Nat. Rev. Drug Discov., 2012, 11(10):751-761. doi: 10.1038/nrd3801

图式 1 四氢苯并[4′, 5′]噻吩并[3′, 2′:5, 6]吡啶并[4, 3-d]嘧啶化合物6~10的合成路线

Scheme 1 Synthetic route of tetrahydrobenzo[4′, 5′]thieno[3′, 2′:5, 6]pyrido[4, 3-d]pyrimidine compounds 6~10

下载: 全尺寸图片幻灯片

表 1 化合物6~10的理化数据及高分辨质谱数据

Table 1. Physical contants and HR-MS of synthesized compounds 6~10

Compd.	R₁	R₂	Yield/%	mp/℃	Physical property	IR (KBr) /cm^-1	HR-MS (ESI)m/z ([M+H]⁺)
Compd.	R₁	R₂	Yield/%	mp/℃	Physical property	IR (KBr) /cm^-1		Calcd.	Found
6a	Ph	Ph	64.2	224~226	White solid	1671 (C=O)	C₂₆H₂₃N₄OS	439.5520	439.5525
6b	Ph	4-OMe-Ph	75.4	218~220	White solid	1678 (C=O)	C₂₇H₂₅N₄O₂S	469.5780	469.5786
6c	Ph	4-F-Ph	64.2	224~226	White solid	1671 (C=O)	C₂₆H₂₂FN₄OS	457.5425	457.5431
6d	Ph	4-Cl-Ph	61.8	234~237	White solid	1684 (C=O)	C₂₆H₂₂ClN₄OS	473.9971	473.9976
6e	Ph	C₃H₇	78.4	215~218	White solid	1672 (C=O)	C₂₃H₂₅N₄OS	405.1749	405.1753
7a	4-OMe-Ph	Ph	68.9	232~234	Yellow solid	1675 (C=O)	C₂₇H₂₅N₄O₂S	469.5780	469.5786
7b	4-OMe-Ph	4-OMe-Ph	72.1	234~237	Yellow solid	1668 (C=O)	C₂₈H₂₇N₄O₃S	499.6040	499.6044
7c	4-OMe-Ph	4-F-Ph	63.6	225~228	White solid	1675 (C=O)	C₂₇H₂₄FN₄O₂S	487.5685	487.5691
7d	4-OMe-Ph	4-Cl-Ph	74.2	244~246	White solid	1669 (C=O)	C₂₇H₂₄ClN₄O₂S	504.0231	504.0236
7e	4-OMe-Ph	C₃H₇	82.5	221~223	White solid	1676 (C=O)	C₂₄H₂₇N₄O₂S	435.5618	435.5622
8a	4-F-Ph	Ph	56.8	226~229	White solid	1672 (C=O)	C₂₆H₂₂FN₄OS	457.5425	457.5431
8b	4-F-Ph	4-OMe-Ph	63.9	223~225	White solid	1670 (C=O)	C₂₇H₂₄FN₄O₂S	487.5685	487.5691
8c	4-F-Ph	4-F-Ph	55.4	226~229	Yellow solid	1672 (C=O)	C₂₆H₂₁F₂N₄OS	475.5329	475.5333
8d	4-F-Ph	4-Cl-Ph	53.2	235~238	Yellow solid	1675 (C=O)	C₂₆H₂₁ClFN₄OS	491.9875	491.9882
8e	4-F-Ph	C₃H₇	53.2	222~225	White solid	1669 (C=O)	C₂₃H₂₄FN₄OS	423.5263	423.5267
9a	4-Cl-Ph	Ph	68.4	242~244	White solid	1678 (C=O)	C₂₆H₂₂ClN₄OS	473.9971	473.9976
9b	4-Cl-Ph	4-OMe-Ph	72.5	243~246	White solid	1671 (C=O)	C₂₇H₂₄ClN₄O₂S	504.0231	504.0236
9c	4-Cl-Ph	4-F-Ph	70.4	240~242	White solid	1677 (C=O)	C₂₆H₂₁ClFN₄OS	491.9875	491.9882
9d	4-Cl-Ph	4-Cl-Ph	67.3	255~257	White solid	1678 (C=O)	C₂₆H₂₁Cl₂N₄OS	508.4421	508.4426
9e	4-Cl-Ph	C₃H₇	61.9	246~249	Yellow solid	1672 (C=O)	C₂₃H₂₄ClN₄OS	439.9809	439.9816
10a	C₃H₇	Ph	57.8	211~213	Yellow solid	1675 (C=O)	C₂₃H₂₅N₄OS	405.1749	405.1753
10b	C₃H₇	4-OMe-Ph	54.6	214~217	Yellow solid	1665 (C=O)	C₂₄H₂₇N₄O₂S	435.5618	435.5622
10c	C₃H₇	4-F-Ph	60.7	212~215	Yellow solid	1668 (C=O)	C₂₃H₂₄FN₄OS	423.5263	423.5267
10d	C₃H₇	4-Cl-Ph	52.9	226~229	Yellow solid	1677 (C=O)	C₂₃H₂₄ClN₄OS	439.9809	439.9816
10e	C₃H₇	C₃H₇	63.7	211~214	Yellow solid	1664 (C=O)	C₂₀H₂₇N₄OS	371.5196	371.5201

下载: 导出CSV

表 2 化合物6~10的¹H NMR和¹³C NMR数据

Table 2. The ¹H NMR and ¹³C NMR data of compounds 6~10

Compd.	¹H NMR (400MHz, CDCl₃) δ	¹³C NMR (101MHz, CDCl₃) δ
6a	7.76 (d, J=8.0Hz, 2H), 7.65 (m, 3H), 7.48 (d, J=8.4Hz, 2H), 7.37 (d, J=8.0Hz, 2H), 7.11 (m, 1H), 3.19 (m, 2H), 3.09 (s, 3H), 2.88 (m, 2H), 1.91 (m, 2H)	164.0, 161.1, 157.5, 151.2, 150.1, 137.8, 136.2, 133.6, 131.2, 130.5, 129.4, 128.2, 127.9, 125.6, 124.4, 122.7, 118.4, 27.2, 26.9, 25.7, 23.1, 22.5
6b	7.79 (d, J=8.0Hz, 2H), 7.41 (d, J=8.0Hz, 2H), 7.12 (d, J=8.0Hz, 2H), 7.07 (m, 1H), 6.97 (d, J=8.4Hz, 2H), 3.79 (s, 3H, OCH₃), 3.21 (m, 2H, CH₂), 3.09 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 2H)	163.7, 163.5, 161.2, 157.4, 152.1, 150.5, 136.9, 133.6, 132.4, 131.1, 129.6, 128.5, 125.7, 124.4, 120.7, 118.8, 116.6, 55.6, 27.2, 26.9, 25.9, 22.7, 22.3
6c	7.81 (d, J=8.8Hz, 2H), 7.83 (d, J=8.4Hz, 2H), 7.52 (d, J=8.0Hz, 2H), 7.29 (d, J=8.4Hz, 2H), 7.14 (m, 1H), 3.21 (m, 2H, CH₂), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.88 (m, 4H)	165.2(d, ¹J_CF =250.4Hz), 163.7, 161.4, 157.9, 151.6, 150.2, 137.9, 136.7, 133.4, 132.2, 130.6 (d, ³J_CF =8.8Hz), 129.7, 128.1(d, ⁴J_CF =4.0Hz), 125.8, 124.4, 116.7 (d, ²J_CF =22.4Hz), 113.1, 27.1, 26.7, 25.8, 22.9, 22.6
6d	7.78 (d, J=8.4Hz, 2H), 7.62 (d, J=8.0Hz, 2H), 7.41 (d, J=8.4Hz, 2H), 7.38 (d, J=8.4Hz, 2H), 7.14 (m, 1H), 3.18 (m, 2H), 3.09 (s, 3H, CH₃), 2.87 (m, 2H), 1.91 (m, 4H)	63.9, 161.3, 158.2, 151.8, 151.1, 137.5, 136.1, 133.2, 132.1, 130.8, 130.1, 128.4, 127.5, 125.8, 124.9, 124.2, 113.1, 27.3, 26.9, 25.6, 22.7, 22.8
6e	7.80 (d, J=8.0Hz, 2H), 7.42 (d, J=8.0Hz, 2H), 7.13 (m, 1H), 3.91 (t, J=7.2Hz, 2H, CH₂), 3.19 (m, 2H), 3.10 (s, 3H, CH₃), 2.88 (m, 2H), 1.88 (m, 4H, CH₂), 1.72 (m, 2H), 0.99 (t, J=6.4Hz, 3H, CH₃)	163.2, 161.2, 157.9, 151.2, 149.5, 136.4, 134.2, 131.6, 128.7, 125.8, 124.4, 122.6, 112.7, 48.8, 27.2, 26.6, 26.5, 22.7, 22.5, 22.1, 11.3
7a	7.74 (d, J=8.0Hz, 2H), 7.61 (d, J=8.4Hz, 2H), 7.41 (d, J=7.6Hz, 2H), 7.15 (m, 1H), 6.98 (d, J=8.0Hz, 2H), 3.78 (s, 3H, OCH₃), 3.21 (m, 2H), 3.11 (s, 3H, CH₃), 2.87 (m, 2H), 1.90 (m, 4H)	163.6, 163.2, 161.4, 158.3, 151.8, 149.7, 137.6, 133.3, 132.5, 130.8, 129.8, 128.4, 127.6, 125.8, 124.9, 124.1, 118.5, 55.7, 27.7, 26.9, 25.7, 22.9, 22.5
7b	7.76 (d, J=8.4Hz, 2H), 7.71 (d, J=8.0Hz, 2H), 7.05 (d, J=7.6Hz, 2H), 6.98 (d, J=8.0Hz, 2H), 3.81 (s, 3H, OCH₃), 3.78 (s, 3H, OCH₃), 3.21 (m, 2H), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 4H)	163.8, 163.6, 163.2, 161.1, 157.9, 151.8, 149.5, 137.8, 136.6, 133.5, 132.4, 130.8, 128.7, 125.3, 124.4, 122.7, 118.8, 55.7, 55.6, 27.9, 26.8, 25.7, 22.9, 22.6
7c	7.80 (d, J=8.4Hz, 2H), 7.72 (d, J=8.0Hz, 2H), 7.21 (d, J=8.0Hz, 2H), 7.02 (d, J=8.0Hz, 2H), 3.79 (s, 3H, OMe), 3.22 (m, CH₂, 2H), 3.11 (s, 3H, CH₃), 2.89 (m, 2H), 1.89 (m, 4H)	165.1(d, ¹J_CF =254.2Hz), 163.5, 163.2, 161.0, 157.9, 151.8, 150.2, 137.9, 136.7, 133.8, 132.4, 130.7 (d, ³J_CF =7.6Hz), 129.4 (d, ⁴J_CF =3.6Hz), 125.7, 124.5, 116.6 (d, ²J_CF =22.8Hz), 113.6, 55.7, 27.9, 26.7, 25.6, 22.9, 22.7
7d	7.78 (d, J=8.0Hz, 2H), 7.65 (d, J=8.4Hz, 2H), 7.39 (d, J=7.2Hz, 2H), 7.01 (d, J=7.6Hz, 2H), 3.79 (s, 3H, OCH₃), 3.20 (m, 2H), 3.12 (s, 3H, CH₃), 2.87 (m, 2H), 1.91 (m, 4H)	163.5, 163.2, 161.2, 159.6, 157.8, 150.3, 148.9, 138.4, 137.4, 132.6, 130.7, 129.6, 129.3, 128.6, 126.3, 124.4, 122.5, 55.9, 27.7, 26.9, 26.5, 22.9, 22.6
7e	7.72 (d, J=8.0Hz, 2H), 6.97 (d, J=7.2Hz, 2H), 3.96 (t, J=6.4Hz, 2H), 3.20 (m, 2H), 3.10 (s, 3H, CH₃), 3.78 (s, 3H, OCH₃), 2.88 (m, 2H), 1.90 (m, 4H), 1.79 (m, 2H), 1.01 (t, J=7.2Hz, 3H, CH₃)	163.3, 161.2, 160.2, 157.9, 151.4, 149.8, 133.5, 131.4, 129.7, 127.1, 125.3, 124.4, 120.5, 55.8, 48.8, 27.9, 27.6, 26.7, 22.9, 22.6, 22.5, 11.3
8a	7.85 (d, J=8.4Hz, 2H), 7.76 (d, J=8.0Hz, 2H), 7.54 (d, J=8.0Hz, 2H), 7.17 (d, J=8.0Hz, 2H), 7.15 (m, 1H), 3.20 (m, 2H, CH₂), 3.10 (s, 3H, CH₃), 2.87 (m, 2H), 1.90 (m, 4H)	165.3(d, ¹J_CF =262.8Hz), 163.4, 161.1, 157.6, 151.2, 149.6, 137.7, 136.5, 133.2, 132.1, 130.7 (d, ³J_CF =9.2Hz), 129.5, 128.4 (d, ⁴J_CF =4.4Hz), 125.9, 124.5, 116.6 (d, ²J_CF =24.0Hz), 113.4, 27.2, 26.9, 25.7, 22.7, 22.5
8b	7.84 (d, J=8.0Hz, 2H), 7.41 (d, J=8.4Hz, 2H), 7.15 (d, J=8.4Hz, 2H), 6.94 (d, J=8.4Hz, 2H), 3.78 (s, 3H, OMe), 3.21 (m, 2H, CH₂), 3.11 (s, 3H, CH₃), 2.88 (m, 2H), 1.91 (m, 2H)	165.3(d, ¹J_CF =261.6Hz), 163.1, 161.2, 157.7, 151.8, 149.6, 137.7, 136.4, 133.7, 132.6, 130.9 (d, ³J_CF =8.4Hz), 129.1, 128.7 (d, ⁴J_CF =4.0Hz), 125.5, 124.4, 111.7 (d, ²J_CF =23.6Hz), 113.6, 55.8, 27.8, 26.9, 25.5, 22.9, 22.6
8c	7.84 (d, J=8.4Hz, 2H), 7.76 (d, J=8.8Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 7.16 (d, J=8.0Hz, 2H), 3.20 (m, 2H, CH₂), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 4H)	165.3(d, ¹J_CF =252.4Hz), 165.1 (d, ¹J_CF =254.8Hz), 163.4, 161.0, 157.5, 151.7, 150.1, 137.9, 132.2, 131.1 (d, ³J_CF =7.2Hz), 130.8 (d, ³J_CF =8.4Hz), 129.4 (d, ⁴J_CF =3.2Hz), 128.6 (d, ⁴J_CF =3.6Hz), 125.5, 117.4 (d, ²J_CF =24.4Hz), 116.6 (d, ²J_CF =23.6Hz), 113.2, 27.5, 26.9, 25.5, 22.8, 22.5
8d	7.86 (d, J=8.0Hz, 2H), 7.82 (d, J=8.8Hz, 2H), 7.18 (d, J=8.4Hz, 2H), 7.15 (d, J=8.0Hz, 2H), 3.21 (m, 2H, CH₂), 3.09 (s, 2H, CH₃), 2.88 (m, 2H), 1.91 (m, 2H)	165.4, 165.2(d, ¹J_CF =254.6Hz), 163.2, 161.1, 157.7, 151.8, 149.5, 137.6, 136.8, 133.2, 132.4, 130.5 (d, ³J_CF =8.4Hz), 129.6 (d, ⁴J_CF =4.0Hz), 125.6, 124.4, 116.9 (d, ²J_CF =23.2Hz), 113.3, 27.5, 26.9, 25.7, 22.9, 22.6
8e	7.85 (d, J=8.0Hz, 2H), 7.16 (d, J=8.0Hz, 2H), 3.93 (t, J=7.2Hz, 2H), 3.20 (m, 2H), 3.09 (s, 3H, CH₃), 2.87 (m, 2H), 1.91 (4H, m), 1.79 (m, 2H), 1.01 (t, J=6.8Hz, 3H, CH₃)	165.2(d, ¹J_CF =236.4Hz), 163.4, 161.3, 157.9, 151.7, 150.1, 137.8, 130.7 (d, ³J_CF =8.0Hz), 129.9, 128.3 (d, ⁴J_CF =4.0Hz), 124.6, 116.5 (d, ²J_CF =22.8Hz), 113.4, 48.8, 27.2, 26.8, 25.9, 22.9, 22.7, 22.5, 11.3
9a	7.80 (d, J=8.4Hz, 2H), 7.68 (d, J=8.0Hz, 2H), 7.54 (d, J=8.0Hz, 2H), 7.46 (d, J=8.4Hz, 2H), 7.15 (m, 1H), 3.22 (m, 2H), 3.12 (s, 3H, CH₃), 2.89 (m, 2H), 1.90 (m, 4H)	163.4, 161.2, 159.4, 157.7, 150.1, 148.8, 138.5, 136.7, 133.4, 131.6, 130.4, 129.8, 129.3, 126.1, 124.2, 122.4, 121.5, 27.2, 26.9, 26.1, 22.9, 22.8
9b	7.81 (d, J=8.8Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 7.11 (d, J=7.6Hz, 2H), 7.04 (d, J=7.6Hz, 2H), 3.78 (s, 3H, OCH₃), 3.21 (m, 2H), 3.10 (s, 3H, CH₃), 2.89 (m, 2H), 1.89 (m, 4H)	163.6, 163.2, 161.2, 159.9, 157.4, 149.6, 148.5, 138.4, 137.2, 132.6, 130.7, 129.8, 129.3, 128.6, 126.0, 124.5, 122.6, 55.9, 27.2, 26.9, 26.3, 22.9, 22.7
9c	7.82 (d, J=8.0Hz, 2H), 7.80 (d, J=8.4Hz, 2H), 7.45 (d, J=8.4Hz, 2H), 7.15 (d, J=8.4Hz, 2H), 3.21 (m, CH₂, 2H), 3.12 (s, 3H, CH₃), 2.88 (m, 2H), 1.91 (m, 4H)	65.4(d, ¹J_CF =225.2Hz), 161.6, 159.7, 157.8, 150.1, 148.9, 138.5, 137.1, 132.3, 130.8 (d, ³J_CF =7.6Hz), 129.5, 129.3, 128.4 (d, ⁴J_CF =4.4Hz), 126.0, 124.6, 116.9 (d, ²J_CF =23.2Hz), 113.6, 27.1, 26.9, 26.5, 22.9, 22.6
9d	7.83 (d, J=8.0Hz, 2H), 7.78 (d, J=8.0Hz, 2H), 7.51 (d, J=7.6Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 3.22 (m, 2H), 3.11 (s, 3H, CH₃), 2.89 (m, 2H), 1.92 (m, 4H)	163.6, 161.0, 159.6, 157.8, 150.1, 148.9, 138.7, 136.9, 133.2, 131.8, 130.2, 129.9, 129.6, 126.5, 124.7, 123.8, 122.5, 27.1, 26.7, 25.8, 22.8, 22.6
9e	7.81 (d, J=8.0Hz, 2H), 7.42 (d, J=8.4Hz, 2H), 3.93 (t, J=7.2Hz, 2H), 3.23 (m, 2H), 3.12 (s, 3H, CH₃), 2.89 (m, 2H), 1.91 (m, 4H), 1.78 (m, 2H), 1.01 (t, J=7.2Hz, 3H)	163.4, 161.3, 159.7, 157.6, 149.7, 148.3, 137.6, 133.8, 130.2, 124.6, 122.4, 118.4, 112.8, 48.9, 27.2, 26.9, 26.1, 22.9, 22.8, 22.5, 11.3
10a	7.86 (d, J=8.4Hz, 2H), 7.46 (d, J=8.0Hz, 2H), 7.12 (m, 1H), 3.94 (t, J=7.6Hz, 2H), 3.20 (m, 2H), 3.10 (s, 3H, CH₃), 2.88 (m, 2H), 1.91 (m, 4H), 1.78 (m, 2H), 0.98 (t, J=7.2Hz, 3H, CH₃)	163.0, 160.5, 157.8, 151.1, 149.4, 136.2, 133.8, 131.1, 128.5, 125.6, 124.7, 122.3, 112.5, 48.4, 27.1, 26.9, 26.1, 22.6, 22.3, 22.1, 11.4
10b	7.73 (d, J=8.4Hz, 2H), 6.96 (d, J=7.6Hz, 2H), 3.97 (t, J=6.8Hz, 2H), 3.18 (m, 2H), 3.11 (s, 3H, CH₃), 3.78 (s, 3H, OCH₃), 2.86 (m, 2H), 1.91 (m, 4H), 1.79 (m, 2H), 1.00 (t, J=7.2Hz, 3H, CH₃)	163.0, 161.4, 160.2, 157.8, 151.4, 150.7, 133.2, 131.4, 129.6, 128.2, 125.1, 123.4, 119.2, 55.8, 48.6, 27.6, 27.1, 26.2, 23.8, 22.6, 22.0, 11.5
10c	7.28 (d, J=8.8Hz, 2H), 7.07 (d, J=8.0Hz, 2H), 3.93 (t, J=7.6Hz, 2H), 3.20 (m, 2H), 3.09 (s, 3H, CH₃), 2.86 (m, 2H), 1.89 (m, 4H), 1.74 (m, 2H), 1.02 (t, J=7.2Hz, 3H, CH₃)	165.4(d, ¹J_CF =228.8Hz), 163.2, 161.2, 159.4, 157.6, 150.4, 149.6, 137.4, 130.2 (d, ³J_CF =8.0Hz), 128.6 (d, ⁴J_CF =3.2Hz), 120.4, 117.8 (d, ²J_CF =22.4Hz), 112.3, 49.1, 27.5, 26.7, 26.1, 23.4, 22.8, 22.6, 11.3
10d	7.47 (d, J=8.0Hz, 2H), 7.23 (d, J=7.6Hz, 2H), 3.94 (t, J=6.4Hz, 2H), 3.17 (m, 2H), 3.09 (s, 3H, CH₃), 2.89 (m, 2H), 1.92 (m, 4H), 1.77 (m, 2H), 0.88 (t, J=7.2Hz, 3H, CH₃)	163.1, 160.8, 157.2, 151.1, 150.2, 136.7, 133.7, 130.0, 128.9, 126.7, 125.6, 122.3, 113.2, 49.4, 27.8, 27.3, 25.9, 23.2, 22.9, 22.5, 11.4
10e	3.95 (t, J=7.2Hz, 2H), 3.21 (m, 2H), 3.21 (m, 2H), 3.09 (s, 3H, CH₃), 2.88 (m, 2H), 1.90 (m, 4H), 1.78 (m, 2H), 1.71 (m, 2H), 1.22 (t, J=7.2Hz, 3H, CH₃), 1.01 (t, J=6.8Hz, 3H, CH₃)	163.2, 161.1, 157.4, 150.8, 149.5, 137.2, 128.3, 125.1, 120.8, 48.9, 47.6, 27.1, 26.9, 25.9, 23.1, 22.9, 22.7, 22.5, 11.6, 11.4

下载: 导出CSV

表 3 化合物6~10的抗肿瘤活性结果(n=3)

Table 3. Whole cell antiproliferative activity of compounds 6~10 (n=3)

Compd.	IC₅₀/(μmol·L^-1)
Compd.	CNE2	KB	MGC-803	MCF-7	PC3
6a	9.7±0.87	10.5±1.03	9.7±0.56	20.1±1.39	17.8±1.44
6b	12.1±1.66	9.9±0.89	8.5±0.42	23.4±1.61	12.6±1.52
6c	8.1±0.74	9.1±0.57	6.4±0.29	18.2±1.72	11.9±0.96
6d	9.7±0.79	11.9±1.31	11.7±1.55	21.8±2.45	20.2±1.73
6e	11.4±1.59	12.1±1.63	14.8±1.92	17.7±1.17	23.5±2.46
7a	11.7±1.85	14.4±2.41	11.5±1.67	24.2±2.32	23.7±2.55
7b	9.6±2.38	12.7±1.49	10.8±2.23	19.1±1.57	20.2±2.78
7c	11.2±1.81	9.9±1.31	8.7±0.78	22.5±1.99	15.5±1.19
7d	13.4±1.46	14.8±1.48	11.6±1.11	11.5±1.22	17.8±2.11
7e	14.8±1.09	15.3±1.76	13.3±1.40	20.8±2.08	13.3±1.71
8a	9.4±0.34	8.6±0.52	7.2±0.66	14.6±1.16	22.5±2.11
8b	8.3±0.77	4.2±0.61	2.4±0.72	17.5±1.63	16.7 ±1.56
8c	3.3±0.54	2.1±0.27	0.9±0.25	9.1±0.99	13.7±1.44
8d	7.9±0.52	3.9±0.50	5.5±0.16	10.6±1.35	17.3±1.27
8e	13.7±0.87	10.5±1.18	9.7±0.93	21.8±2.67	22.5±1.89
9a	14.3±1.17	12.8±1.15	7.8±0.94	17.2±2.27	15.9±1.65
9b	12.4±1.93	9.9±0.58	8.6±0.83	14.4±1.10	13.4±1.33
9c	7.8±0.56	6.4±0.76	4.2±0.45	8.9±0.76	16.3±2.77
9d	9.3±0.82	7.8±0.69	6.9±0.42	13.8±1.52	11.4±1.82
9e	11.5±1.79	10.1±1.76	11.7±2.11	21.7±2.43	17.3±1.35
10a	18.4±2.25	16.5±1.43	16.4±1.73	18.7±2.14	16.6±1.71
10b	21.7±1.84	25.3±2. 74	18.9±1.04	16.4±1.57	21.3±2.18
10c	14.4±1.01	11.5±1.62	14.8±1.14	13.8±1.79	18.2±1.64
10d	11.9±1.47	17.8±1.13	15.1±1.29	18.1±1.35	17.7±1.51
10e	25.2±1.51	22.3±2.14	19.3±2.55	15.8±1.29	21.6±2.26
5-FU	17.8±1.36	12.5±0.87	18.4±1.43	10.8±0.96	15.4±1.22

下载: 导出CSV

表 4 化合物6a~6c、7c、8a~8e、9a~9d和5-FU的体外治疗指数(IVTI)

Table 4. In vitro therapeutic index of compounds 6a~6c, 7c, 8a~8e, 9a~9d and 5-FU

Compd.	MGC-803	GES-1	IVTI
6a	9.7±0.56	>100	>10.31
6b	8.5±0.42	>100	>11.76
6c	6.4±0.29	>100	>15.62
7c	8.7±0.78	>100	>11.49
8a	7.2±0.66	>100	>13.88
8b	2.4±0.72	>100	>41.66
8c	0.9±0.25	>100	>111.11
8d	5.5±0.16	>100	>18.18
8e	9.7±0.93	>100	>10.30
9a	7.8±0.94	>100	>12.82
9b	8.6±0.83	>100	>11.62
9c	4.2±0.45	>100	>23.80
9d	6.9±0.42	>100	>14.49
5-FU	18.4±1.43	10.7±0.59	0.58
IVTI-The ratio of IC₅₀(GES-1) and IC₅₀(MGC-803)

下载: 导出CSV

扫一扫看文章

计量

PDF下载量: 0
文章访问数: 0
HTML全文浏览量: 0

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