药学学报


H6a	H	3-H	61.92	200 >	59.58	78.82
H6b	H	3-Methoxy	157.3	200 >	35.59	142
H6c	H	4-Methoxy	282.6	200 >	42.65	66.44
H6d	H	3, 5-Dimethoxy	200 >	200 >	200 >	200 >
H6e	H	3-Chloro	41.77	82.83	42.65	40.68
H6f	H	4-Chloro	29.09	86.67	33.38	41.63
H6g	H	3-Fluoro	52.53	83.85	27.81	49.51
H6h	H	4-COOC₂H₅	200 >	147.4	10.88	17.61
F6a	Fluoro	3-H	143.4	81.22	58.78	144.9
F6b	Fluoro	3-Methoxy	200 >	200 >	118.3	200 >
F6c	Fluoro	4-Methoxy	200 >	120.2	78.57	141.9
F6d	Fluoro	3, 5-Dimethoxy	126.6	105.8	200 >	200 >
F6e	Fluoro	3-Chloro	152.9	200 >	142.3	200 >
F6f	Fluoro	4-Chloro	137.4	200 >	79.19	183.1
F6g	Fluoro	3-Fluoro	126	200	200 >	200 >
F6h	Fluoro	4-COOC₂H₅	200 >	200 >	200 >	200 >
LY294002	-	-	23.60	20.61	11.80	11.41


H6a	H	3-H	61.92	200 >	59.58	78.82
H6b	H	3-Methoxy	157.3	200 >	35.59	142
H6c	H	4-Methoxy	282.6	200 >	42.65	66.44
H6d	H	3, 5-Dimethoxy	200 >	200 >	200 >	200 >
H6e	H	3-Chloro	41.77	82.83	42.65	40.68
H6f	H	4-Chloro	29.09	86.67	33.38	41.63
H6g	H	3-Fluoro	52.53	83.85	27.81	49.51
H6h	H	4-COOC₂H₅	200 >	147.4	10.88	17.61
F6a	Fluoro	3-H	143.4	81.22	58.78	144.9
F6b	Fluoro	3-Methoxy	200 >	200 >	118.3	200 >
F6c	Fluoro	4-Methoxy	200 >	120.2	78.57	141.9
F6d	Fluoro	3, 5-Dimethoxy	126.6	105.8	200 >	200 >
F6e	Fluoro	3-Chloro	152.9	200 >	142.3	200 >
F6f	Fluoro	4-Chloro	137.4	200 >	79.19	183.1
F6g	Fluoro	3-Fluoro	126	200	200 >	200 >
F6h	Fluoro	4-COOC₂H₅	200 >	200 >	200 >	200 >
LY294002	-	-	23.60	20.61	11.80	11.41


P6a	H	80.23	30.89	20.15	20.54
P6b	3-Methoxy	17.52	11.12	3.241	24.31
P6c	4-Methoxy	60.33	11.56	6.199	40.96
P6d	3, 5-Dimethoxy	6.004	0.228 7	4.622	11.29
P6e	3-Chloro	29.07	18.01	4.775	18.92
P6f	3-Fluoro	189.2	36.74	4.503	27.63
LY294002	-	23.60	20.61	11.80	11.41


P6a	H	80.23	30.89	20.15	20.54
P6b	3-Methoxy	17.52	11.12	3.241	24.31
P6c	4-Methoxy	60.33	11.56	6.199	40.96
P6d	3, 5-Dimethoxy	6.004	0.228 7	4.622	11.29
P6e	3-Chloro	29.07	18.01	4.775	18.92
P6f	3-Fluoro	189.2	36.74	4.503	27.63
LY294002	-	23.60	20.61	11.80	11.41

基于喹喔啉为母核的PI3K抑制剂的合成及抗肿瘤活性研究

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黄涛 ¹^,² , 赖惠芳 ² , 林荣坤 ² , 林锦 ² , 李柱来 ²^,^* , 许秀枝 ²^,^*

药学学报 | 研究论文 2020,55(1): 96-105

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药学学报 | 研究论文 2020, 55(1): 96-105

基于喹喔啉为母核的PI3K抑制剂的合成及抗肿瘤活性研究

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黄涛¹^,², 赖惠芳², 林荣坤², 林锦², 李柱来²^,^*, 许秀枝²^,^*

作者信息

1.福建省儿童医院(福建省妇幼保健院), 福建福州 350001

2.福建医科大学药学院, 福建省药物靶点发现与结构功能研究重点实验室, 福建福州 350122

通讯作者:

*李柱来, E-mail:lizhulai@126.com;

许秀枝, Tel:86-591-22862016, E-mail:xiuzhi_xu@126.com

Synthesis and biological evaluation of phosphoinositide 3-kinase (PI3K) inhibitors based on a quinoxaline scaffold

Tao HUANG¹^,², Hui-fang LAI², Rong-kun LIN², Jin LIN², Zhu-lai LI²^,^*, Xiu-zhi XU²^,^*

Affiliations

1. Fujian Provincial Children's Hospital(Fujian Provincial Maternity and Children's Hospital), Fuzhou 350001, China

2. Fujian Key Laboratory of Drug Target Discovery and Structural and Functional Research, College of Pharmacy, Fujian Medical University, Fuzhou 350122, China

出版时间: 2020-01-12 doi: 10.16438/j.0513-4870.2019-0679

文章导航

摘要

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以含有喹喔啉母核的PI3K抑制剂XL765和WR23为结构基础，通过生物电子等排，在喹喔啉母核上2位引入取代苯氧基片段，延长3位连接链改为磺酰肼，并在7位引入氟原子；去掉3位取代并在7位引入丙烯酰胺基。初步设计并合成了22个喹喔啉类衍生物，通过¹H NMR、¹³C NMR、ESI-MS进行结构确证。以人非小细胞肺癌A549、人乳腺癌细胞MCF-7、人结肠癌细胞HCT-116和人肝癌细胞HepG2进行体外抗肿瘤活性筛选（MTT法）。结果表明，P6b、P6e、P6f对HCT116活性较好（IC₅₀=3.24，4.78和4.50 μmol·L^-1），P6d对MCF-7具有较强抑制作用（IC₅₀=0.228 7 μmol·L^-1）。

关键词

喹喔啉类衍生物 / 抗肿瘤 / PI3K抑制剂

Abstract

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Based on the structure of inhibitors XL765 and WR23, the quinoxaline scaffold was selected as an attractive structure for drug design. In this protocol, the 2-position of quinoxaline was modified with a substituted phenoxy fragment. Meanwhile, the linking chain at the 3-position was changed to a sulfonyl hydrazine or was removed. A series of substituent groups were added at the 6-position of the quinoxaline scaffold. Twenty-two quinoline derivatives were designed and synthesized, and their structures were confirmed by ¹H NMR, ¹³C NMR, and ESI-MS. All compounds were screened for anti-tumor activity in vitro in A549, MCF-7, HCT-116 and HepG2 cancer cells. The results showed that P6b was effective, P6e and P6f had better activity against HCT116 (IC₅₀=3.24, 4.78 and 4.50 μmol·L^-1), and P6d had strong inhibitory effect on MCF-7 (IC₅₀=0.228 7 μmol·L^-1).

Key words

quinoxaline derivative / anti-tumour / PI3K inhibitor

引用本文

黄涛, 赖惠芳, 林荣坤, 林锦, 李柱来, 许秀枝. 基于喹喔啉为母核的PI3K抑制剂的合成及抗肿瘤活性研究. 药学学报, 2020 , 55 (1) : 96 -105 . DOI: 10.16438/j.0513-4870.2019-0679

Tao HUANG, Hui-fang LAI, Rong-kun LIN, Jin LIN, Zhu-lai LI, Xiu-zhi XU. Synthesis and biological evaluation of phosphoinositide 3-kinase (PI3K) inhibitors based on a quinoxaline scaffold[J]. Acta Pharmaceutica Sinica, 2020 , 55 (1) : 96 -105 . DOI: 10.16438/j.0513-4870.2019-0679

正文

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PI3K/Akt/mTOR信号通路是由磷脂酰肌醇-3激酶(PI3Ks)及其下游介质Akt和哺乳动物雷帕霉素靶蛋白(mTOR)构成PI3K/Akt/mTOR信号级联、调节细胞增殖、生存和代谢的核心成分^[1], 是连接癌基因和多种受体与许多重要细胞功能的关键信号转导系统, 可能是人类癌症中最常激活的信号通路^[2]。磷脂酰肌醇3-激酶(PI3Ks)是PI3K/Akt/mTOR信号通道中的关键激酶, 它整合来自生长因子、细胞因子和其他环境线索的信号, 将其转化为调节多种信号通路的细胞内信号。这些通路控制许多生理功能和细胞过程, 包括细胞增殖、生长、存活、运动和代谢, 且PI3K的激活常见于各种癌症中^{[2, 3]}。PI3Ks作为酪氨酸激酶受体(RTKs)和G蛋白偶联受体(GPCRs)下游的主要效应物, 是PI3K/Akt/mTOR信号通道中关键性激酶, 能够使磷脂酰肌醇4, 5-二磷酸(PIP2)的3′-羟基磷酸化生成磷脂酰肌醇3, 4, 5-三磷酸(PIP3), 这种脂质产物作为“第二信使”将各种生长因子和细胞因子的信号转导到细胞内, 进而激活丝氨酸/苏氨酸激酶Akt等下游效应通路^[4]。最终作用细胞核内翻译起始因子4E-BP、S6Ks, 进而影响细胞的翻译、转录、生长和增殖。

LY294002 (图 1)是第一个可逆的靶向PI3K家族的小分子抑制剂(对PI3Kα、PI3Kδ和PI3Kβ半数抑制率IC₅₀分别为0.50、0.57、0.97 μmol·L^-1)。但是两者对单个PI3K亚型几乎没有选择性, 在动物实验中也具有相当大的毒性^{[5, 6]}。XL765 (图 1)是一个含喹喔啉母环的高选择性的强ATP竞争性, 可逆泛I类PI3K抑制剂和mTOR双重抑制剂^[7]。它作用于p110α、p110β、p110δ、p110γ和mTOR的IC₅₀分别为39、113、9、43、157 nmol·L^-1。XL765的药效学活性与抑制迁移性肿瘤细胞的增殖、抑制肿瘤血管生成、诱导细胞凋亡有关, 在多种人体肿瘤模型中均有明显的肿瘤生长抑制作用^{[8, 9]}。WR23 (图 1)也是含有喹喔啉母核的PI3K抑制剂, 它是由Wu等^[10-12]发现的含有吗啉环喹喔啉类PI3Kα抑制剂, 对PI3Kα半数抑制率IC₅₀为0.025 μmol·L^-1。Corona等^[13]研究表明, 根据电子等排体将2位链接分别替换为O、S时, 同时在60种人类癌细胞上分别测试2位链接为N、O、S 3种类型的新化合物的抑制活性, 研究结果显示O链接活性表现最高。本文以喹喔啉为母核, 以PI3K为作用靶点, 设计并合成了22个喹喔啉类衍生物, 通过¹H NMR、¹³C NMR、ESI-MS进行结构确证, 并对其进行抗肿瘤活性筛选, 初步探究其构效关系, 为该类化合物作为PI3K抑制剂进一步的探索及其结构优化研究提供依据。

结果与讨论

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1 化学合成

以邻苯二胺和4-氟邻苯二胺为起始原料, 与草酸二乙酯环合得到喹喔啉酮(H2)和6-氟喹喔啉酮(F2), 经三氯氧磷氯化得到2, 3-二氯喹喔啉(H3)和6-氟-2, 3-二氯喹喔啉(F3), 再与取代苯酚在弱碱性条件下发生亲核取代反应生成2-苯氧基类喹喔啉衍生物(H4a~H4h, F4a~F4h), 与水合肼反应后再与对甲苯磺酰氯酰化最终得到16个磺酰肼类喹喔啉衍生物(H6a~H6h, F6a~F6h) (合成路线1)。以4-硝基邻苯二胺为起始原料, 经三氯氧磷氯化和取代苯酚的亲核取代反应得到6-硝基-3-取代苯氧基喹喔啉衍生物(P4a~P4f), 再经Pd/C/H₂还原后与丙烯酰氯反应得到6个丙烯酰胺类喹喔啉衍生物(P6a~P6f) (合成路线2)。目标化合物经¹H NMR、¹³C NMR和ESI-MS确证其结构, 数据见实验部分。

2 生物活性评价

以人非小细胞肺癌A549、人乳腺癌细胞MCF-7、人结肠癌细胞HCT-116、人肝癌细胞Hep-G2为实验对象, 通过MTT法对所得22个目标化合物进行体外抗肿瘤细胞增殖实验, 并以LY294002为阳性对照, 结果见表 1、2和图 2。其中P6b、P6e、P6f对HCT116活性较好(IC₅₀=3.24、4.78和4.50 μmol·L^-1), P6d对MCF-7具有较强抑制作用(IC₅₀=0.228 7 μmol·L^-1)。

3 分子对接

利用SYBYL/Surflex-Dock模块, 以PI3Kα和抑制剂GCD-0326的晶体复合物(PDB: 5DXT)的蛋白数据为基础, 对化合物进行分子模拟对接进行简单初步分析。首先对配体分子进行Tripos力场能量最小化计算, 最大重复次数(Max Iterations)设置为1 000, Gradient能量差设置为0.005, 得到最低能量构象的小分子用于对接分析。结果表明, 化合物都能与靶点位置形成较好的氢键作用, P6d对接示意图显示(图 3), 苯氧基上3, 5-二甲氧基分别与Lys802、Tyr836形成氢键; 喹喔啉母核与丙烯酰胺分别与Ser774、Thr856形成氢键作用。根据活性筛选结果以及对接结果可得出, 苯氧基上3, 5-二甲氧基取代能明显提高活性, 丙烯酰胺的引入也对活性有一定的影响。

4 小结

本文以喹喔啉母核为结构基础, 在喹喔啉母核上2位引入取代苯氧基片段, 延长3位连接链改为磺酰肼或去掉3位取代, 并在6位加入一系列取代基团, 设计并合成了22个目标化合物, 并通过¹H NMR、¹³C NMR、ESI-MS进行了确证。采用MTT法进行抗肿瘤活性测试, 结果显示丙烯酰胺类喹喔啉衍生物具有相对较好的活性, 并以3, 5-二甲氧基取代(P6d)活性最佳(IC₅₀=0.228 7 μmol·L^-1)。利用计算机辅助药物设计进行分子对接分析, 结果显示目标化合物与作用靶点具有较好的结合作用, 生物活性实验也证实了其具有良好的相关性。本研究结果表明, 该类化合物可进行进一步的探索及其结构优化研究, 对针对PI3K信号通路的小分子抗肿瘤药物的开发具有一定的意义和价值。

实验部分

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RE52-98型旋转蒸发仪; WRS-1B数字熔点仪; 核磁共振波谱仪: Bruker AVANCE Ⅲ 400M、Bruker ascend 500M、Bruker AVANCE ⅢHD 600 MHz; 质谱仪: Thermo Finnigan; Thermol Labsystems酶标仪: Multiskan Mk3; 4-硝基邻苯二胺(G1517071)、苯酚(H1820089)、3-氯苯酚(L1712072)、4-氯苯酚(J1631082)、3-氟苯酚(G1528032)、尼泊金乙酯(K1715100)、对甲基苯磺酰氯(L1606048)、草酸二乙酯(F1526034), 阿拉丁生化科技股份有限公司; 4-氟邻苯二胺(C10078939), 上海麦克林生化科技有限公司; 水合肼(20160510)、三氯氧磷(20180111)、N, N-二甲基甲酰胺(20180117), 国药集团药业股份有限公司; 3-甲氧基苯酚(M71420GGQ0), 天津希恩思生化科技有限公司; 4-甲氧基苯酚(EH030124)、丙烯酰氯(GD270002)、3, 5-二甲氧基苯酚(FJ210258), 萨恩化学技术(上海)有限公司。

1 化合物的合成

4-1-1 1, 4-二氢喹喔啉-2, 3-二酮(H2)的合成

邻苯二胺100 mmol、草酸二乙酯100 mmol和4 mol·L^-1盐酸水溶液80 mL, 90 ℃搅拌6 h。反应液静置冷却析出紫红色晶体, 抽滤, 水洗, 干燥, 得紫红色晶体14.29 g, 收率88%。同法, 用4-氟邻苯二胺代替邻苯二胺合成F2, 得亮黑色晶体, 收率92%。同法用4-硝基邻苯二胺替代邻苯二胺得到P2, 紫红色固体, 收率95%。

1.2 2, 3-二氯-喹喔啉(H3)的合成

1, 4-二氢喹喔啉-2, 3-二酮(H2) 60 mmol、新蒸POCl₃ 120 mL、10 mL DMF, 回流搅拌4 h。反应液倒入800 mL冰水中, 析出大量灰色固体, 抽滤, 水洗, 干燥后得灰色固体, 正丁醇重结晶得白色固体粉末10.77 g, 收率90%。

同法合成F3, 白色固体粉末, 收率91%; P3, 白色固体粉末, 收率92%。

1.3 2-氯-3-苯氧基喹喔啉(H4a)的合成

2, 3-二氯喹喔啉(H3) 6.00 mmol溶于10 mL DMF中, 加K₂CO₃ 7.20 mmol, 滴加入苯酚(6 mmol)的DMF (8 mL)溶液, 70 ℃反应1 h。反应液倒入40 mL饱和食盐水中, 静置, 过滤, 乙醇重结晶得浅黄色固体0.85 g, 收率55%。

同法合成H4b~H4h、F4a~F4h、P4a~P4f。

1.4 2-肼基-3-苯氧基喹喔啉(H5a)的合成

2-氯-3-苯氧基喹喔啉2.00 mmol (H4a)、80 mL乙醇、1.5 mL 85%水合肼, 常温搅拌24 h, 过滤得棕黄色固体0.46 mg, 收率91%。同法合成H5b~H5h、F5a~F5h。

4-1-5 N'-(3-苯氧基喹喔啉)-4-甲基苯磺酰肼(H6a)的合成

2-肼基-3-苯氧基喹喔啉(H5a) 1.55 mmol溶于30 mL二氯甲烷中, 加入对甲苯磺酰氯1.86 mmol, 吡啶两滴, 常温搅拌4 h, 反应液拌硅胶, 硅胶柱分离, 得浅黄色固体0.41 g, 收率65%, mp: 173.6~175.1 ℃; ESI-MS (m/z): 405.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.95 (s, 1H), 9.84 (s, 1H), 7.77 (d, J = 8.2 Hz, 2H), 7.53~7.45 (m, 2H), 7.39 (dd, J = 12.1, 5.6 Hz, 2H), 7.32 (d, J = 7.8 Hz, 3H), 7.26 (t, J = 7.7 Hz, 2H), 7.22 (d, J = 8.3 Hz, 2H), 2.15 (s, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 152.60, 147.59, 143.44, 143.41, 138.37, 137.78, 135.20, 130.16, 129.38, 128.28, 127.54, 126.60, 126.01, 125.84, 125.69, 122.31, 21.26。

同法合成H6b~H6h、F6a~F6h。

N'-(3-(3-甲氧基苯氧基)喹喔啉-2-基)-4-甲基-苯磺酰肼(H6b), 浅黄色固体, 收率63%, mp: 159.3~161.7 ℃; ESI-MS (m/z): 435.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.95 (d, J = 1.8 Hz, 1H), 9.83 (s, 1H), 7.78 (d, J = 8.2 Hz, 2H), 7.42 (d, J = 7.8 Hz, 2H), 7.38 (d, J = 8.2 Hz, 1H), 7.30 (d, J = 7.3 Hz, 1H), 7.24 (dd, J = 12.5, 8.1 Hz, 3H), 6.91 (dd, J = 14.1, 5.3 Hz, 3H), 3.79 (s, 3H), 2.17 (s, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 160.72, 153.62, 147.47, 143.43, 143.41, 138.38, 137.81, 135.25, 130.55, 129.37, 128.29, 127.56, 126.66, 125.84, 125.70, 114.34, 111.69, 108.26, 55.86, 21.26。

N'-(3-(4-甲氧基苯氧基)喹喔啉-2-基)-4-甲基-苯磺酰肼(H6c), 浅黄色固体, 收率65%, mp: 177.0~178.6 ℃; ESI-MS (m/z): 435.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.93 (s, 1H), 9.81 (s, 1H), 7.78 (d, J = 8.2 Hz, 2H), 7.38 (d, J = 7.8 Hz, 2H), 7.29 (d, J = 7.9 Hz, 1H), 7.24 (dd, J = 14.1, 5.4 Hz, 5H), 7.03 (d, J = 9.0 Hz, 2H), 3.80 (s, 3H), 2.16 (s, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 157.12, 147.95, 145.79, 143.41, 143.40, 138.28, 137.79, 135.29, 129.37, 128.28, 127.38, 126.53, 125.81, 125.64, 123.27, 115.03, 55.87, 21.26。

N'-(3-(3, 5-甲氧基苯氧基)喹喔啉-2-基)-4-甲基-苯磺酰肼(H6d), 浅黄色固体, 收率65%, mp: 135.1~137.4 ℃; ESI-MS (m/z): 465.15 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.95 (s, 1H), 9.81 (s, 1H), 7.78 (d, J = 8.2 Hz, 2H), 7.45 (d, J = 8.1 Hz, 1H), 7.41 (d, J = 6.9 Hz, 1H), 7.30 (t, J = 7.6 Hz, 1H), 7.25 (d, J = 9.8 Hz, 2H), 7.22 (s, 1H), 6.54 (d, J = 2.1 Hz, 2H), 6.46 (t, J = 2.1 Hz, 1H), 3.76 (s, 6H), 2.17 (s, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 161.34, 154.24, 147.35, 143.40, 143.39, 138.37, 137.85, 135.26, 129.37, 128.28, 127.58, 126.72, 125.82, 125.70, 100.81, 98.01, 55.96, 21.27。

N'-(3-(3-氯苯氧基)喹喔啉-2-基)-4-甲基-苯磺酰肼(H6e), 浅黄色固体, 收率67%, mp: 148.7~150.9 ℃; ESI-MS (m/z): 439.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.99 (d, J = 2.0 Hz, 1H), 9.86 (d, J = 1.7 Hz, 1H), 7.78 (d, J = 8.2 Hz, 2H), 7.57~7.50 (m, 2H), 7.47~7.38 (m, 3H), 7.36 (dd, J = 8.1, 2.2 Hz, 1H), 7.33~7.28 (m, 1H), 7.28~7.24 (m, 1H), 7.22 (d, J = 8.1 Hz, 2H), 2.16 (s, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 153.32, 147.18, 143.41, 143.32, 138.53, 137.81, 135.03, 133.91, 131.55, 129.38, 128.29, 127.77, 126.70, 126.13, 125.86, 125.76, 122.61, 121.24, 21.26。

N'-(3-(4-氯苯氧基)喹喔啉-2-基)-4-甲基-苯磺酰肼(H6f), 浅黄色固体, 收率62%, mp: 189.2~189.8 ℃; ESI-MS (m/z): 439.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.96 (d, J = 2.1 Hz, 1H), 9.85 (d, J = 2.0 Hz, 1H), 7.76 (d, J = 8.2 Hz, 2H), 7.56~7.52 (m, 2H), 7.40 (s, 1H), 7.39 (d, J = 2.4 Hz, 2H), 7.37 (s, 1H), 7.31~7.26 (m, 1H), 7.24 (d, J = 8.3 Hz, 1H), 7.20 (d, J = 8.1 Hz, 2H), 2.14 (s, 3H); ¹³CNMR (151 MHz, DMSO-d₆) δ 151.35, 147.32, 143.42, 143.34, 138.46, 137.79, 135.06, 130.05, 129.38, 128.28, 127.70, 126.64, 125.85, 125.75, 124.27, 21.26。

N'-(3-(3-氟苯氧基)喹喔啉-2-基)-4-甲基-苯磺酰肼(H6g), 浅黄色固体, 收率68%, mp: 146.3~148.5 ℃; ESI-MS (m/z): 424.10 [M]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.98 (d, J = 1.9 Hz, 1H), 9.86 (d, J = 1.8 Hz, 1H), 7.78 (d, J = 8.2 Hz, 2H), 7.54 (dd, J = 15.2, 8.2 Hz, 1H), 7.43 (t, J = 7.4 Hz, 2H), 7.37~7.28 (m, 2H), 7.26 (d, J = 7.8 Hz, 1H), 7.22 (d, J = 8.1 Hz, 3H), 7.20~7.14 (m, 1H), 2.16 (s, 3H); ¹³CNMR (126 MHz, DMSO-d₆) δ 162.80 (d, J = 244.6 Hz), 153.60 (d, J = 11.3 Hz), 147.15, 143.42, 143.36, 138.54, 137.83, 135.08, 131.36 (d, J = 9.5 Hz), 129.39, 128.29, 127.78, 126.73, 125.87, 125.77, 118.49 (d, J = 3.0 Hz), 112.95 (d, J = 20.9 Hz), 110.21 (d, J = 24.5 Hz), 21.25。

4-((3-对甲苯磺酰肼)喹喔啉-2-基)氧基苯甲酸乙酯(H6h), 白色固体, 收率65%, mp: 154.0~156.1 ℃; ESI-MS (m/z): 477.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 10.00 (d, J = 2.2 Hz, 1H), 9.92 (d, J = 2.1 Hz, 1H), 8.09 (d, J = 8.7 Hz, 2H), 7.78 (d, J = 8.2 Hz, 2H), 7.51 (d, J = 8.7 Hz, 2H), 7.48~7.39 (m, 2H), 7.31 (t, J = 7.6 Hz, 1H), 7.27 (d, J = 8.2 Hz, 1H), 7.22 (d, J = 8.1 Hz, 2H), 4.35 (q, J = 7.1 Hz, 2H), 2.16 (s, 3H), 1.35 (t, J = 7.1 Hz, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 165.55, 156.49, 146.96, 143.43, 138.59, 137.77, 134.98, 132.16, 131.41, 129.38, 129.13, 128.28, 127.90, 127.43, 126.72, 122.42, 61.30, 21.26, 14.66。

N'-(3-苯氧基-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6a), 白色固体, 收率70%, mp: 183.5~184.4 ℃; ESI-MS (m/z): 423.00 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.97 (s, 1H), 9.87 (s, 1H), 7.77 (d, J = 8.2 Hz, 2H), 7.50 (t, J = 7.8 Hz, 2H), 7.33 (d, J = 7.2 Hz, 4H), 7.27 (d, J = 6.1 Hz, 1H), 7.24 (s, 1H), 7.23~7.17 (m, 2H), 2.17 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 159.80 (d, J = 242.0 Hz), 152.46, 148.39, 143.43, 143.08, 137.82, 135.78 (d, J = 13.0 Hz), 135.30, 130.20, 129.38, 128.29, 127.43 (d, J = 9.7 Hz), 126.17, 122.30, 116.29 (d, J = 24.0 Hz), 111.14 (d, J = 22.8 Hz), 21.26。

N'-(3-(3-甲氧基苯氧基)-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6b), 白色固体, 收率75%, mp: 177.6~180.4 ℃; ESI-MS (m/z): 453.05 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.93 (s, 1H), 9.83 (s, 1H), 7.78 (d, J = 7.5 Hz, 2H), 7.40 (t, J = 8.0 Hz, 1H), 7.36~7.30 (m, 1H), 7.30~7.26 (m, 1H), 7.24 (d, J = 7.3 Hz, 3H), 6.97~6.86 (m, 3H), 3.80 (s, 3H), 2.19 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 159.81 (d, J = 239.0 Hz), 153.46, 148.28, 143.42, 143.07, 137.85, 135.82 (d, J = 12.8 Hz), 135.32, 130.59, 129.38, 129.13, 128.30, 127.42 (d, J = 9.7 Hz), 116.31 (d, J = 24.3 Hz), 114.32, 111.90, 111.20 (d, J = 22.4 Hz), 108.26, 55.89, 21.25。

N'-(3-(4-甲氧基苯氧基)-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6c), 白色固体, 收率72%, mp: 171.1~172.8 ℃; ESI-MS (m/z): 453.05 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.92 (s, 1H), 9.83 (s, 1H), 7.78 (d, J = 7.6 Hz, 2H), 7.29 (d, J = 8.0 Hz, 3H), 7.26 (d, J = 13.3 Hz, 2H), 7.20 (d, J = 9.3 Hz, 4H), 2.36 (s, 3H), 2.18 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 159.80 (d, J = 241.8 Hz), 150.18, 148.56, 143.41, 143.07, 137.82, 135.85 (d, J = 12.8 Hz), 135.32, 135.25, 130.54, 129.37, 129.13, 128.29, 127.40 (d, J = 9.8 Hz), 122.05, 116.17 (d, J = 24.5 Hz), 111.10 (d, J = 22.5 Hz), 20.92。

N'-(3-(3, 5-二甲氧基苯氧基)-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6d), 白色固体, 收率75%, mp: 186.4~187.1 ℃; ESI-MS (m/z): 483.10 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.95 (s, 1H), 9.82 (s, 1H), 7.77 (d, J = 8.1 Hz, 2H), 7.32 (t, J = 7.6 Hz, 1H), 7.27 (dd, J = 13.2, 7.4 Hz, 2H), 7.23 (d, J = 8.0 Hz, 2H), 6.54 (s, 2H), 6.46 (s, 1H), 3.76 (s, 6H), 2.19 (s, 3H); ¹³C NMR (151 MHz, DMSO-d₆) δ 161.37, 159.80 (d, J = 242.2 Hz), 154.05, 148.16, 143.40, 143.01, 137.88, 135.78, 135.30, 129.37, 128.29, 127.41, 116.40, 111.34, 100.82, 98.18, 55.97, 21.26。

N'-(3-(3-氯苯氧基)-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6e), 白色固体, 收率70%, mp: 186.1~186.7 ℃; ESI-MS (m/z): 457.00 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.96 (s, 1H), 7.78 (d, J = 7.3 Hz, 2H), 7.57~7.50 (m, 2H), 7.41 (d, J = 8.0 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.33 (d, J = 8.3 Hz, 1H), 7.27 (t, J = 7.9 Hz, 2H), 7.23 (d, J = 7.7 Hz, 2H), 2.18 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 159.83 (d, J = 242.2 Hz), 153.17, 148.02, 143.43, 142.97, 137.84, 135.62 (d, J = 12.9 Hz), 135.49, 133.97, 131.58, 129.38, 128.31, 127.46 (d, J = 9.8 Hz), 126.30, 122.65, 121.26, 116.54 (d, J = 24.5 Hz), 111.25 (d, J = 22.5 Hz), 21.25。

N'-(3-(4-氯苯氧基)-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6f), 白色固体, 收率70%, mp: 199.4~200.2 ℃; ESI-MS (m/z): 457.00 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.95 (s, 1H), 9.87 (s, 1H), 7.78 (d, J = 7.9 Hz, 2H), 7.56 (d, J = 8.6 Hz, 2H), 7.41 (d, J = 8.6 Hz, 2H), 7.33 (t, J = 8.7 Hz, 1H), 7.29 (d, J = 6.2 Hz, 1H), 7.23 (d, J = 7.8 Hz, 3H), 2.18 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 159.83 (d, J = 242.0 Hz), 151.21, 148.14, 143.43, 142.98, 137.82, 135.64 (d, J = 12.7 Hz), 135.42, 130.24, 130.10, 129.38, 128.30 (s), 127.46 (d, J = 9.8 Hz), 124.28, 116.46 (d, J = 24.5 Hz), 111.18 (d, J = 22.9 Hz), 21.25。

N'-(3-(3-氟苯氧基)-7-氟喹喔啉-2-基)-4-甲基-苯磺酰肼(F6g), 白色固体, 收率72%, mp: 174.2~175.6 ℃; ESI-MS (m/z): 441.00 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.99 (s, 1H), 9.88 (s, 1H), 7.77 (d, J = 7.9 Hz, 2H), 7.54 (dd, J = 15.3, 8.1 Hz, 1H), 7.34 (t, J = 9.8 Hz, 2H), 7.27 (d, J = 5.1 Hz, 1H), 7.26~7.15 (m, 5H), 2.18 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 162.80 (d, J = 244.7 Hz), 159.84 (d, J = 242.2 Hz), 153.40 (d, J = 11.3 Hz), 147.97, 143.43, 142.96, 137.85, 135.63 (d, J = 12.8 Hz), 135.47, 131.40 (d, J = 9.6 Hz), 129.39, 128.30, 127.46 (d, J = 9.9 Hz), 118.51 (d, J = 3.1 Hz), 116.54 (d, J = 24.4 Hz), 113.13 (d, J = 21.0 Hz), 111.25 (d, J = 22.5 Hz), 110.27 (d, J = 24.6 Hz), 21.25。

4-甲基-((3-(2-苯磺酰肼)-6-氟喹喔啉-2-基)-4-氧基)苯磺酰肼(F6h), 白色固体, 收率70%, mp: 172.2~173.8 ℃; ESI-MS (m/z): 495.05 [M-H]^－; ¹H NMR (400 MHz, DMSO-d₆) δ 9.97 (s, 1H), 9.92 (s, 1H), 8.09 (d, J = 7.8 Hz, 2H), 7.78 (d, J = 7.5 Hz, 2H), 7.52 (d, J = 7.6 Hz, 2H), 7.34 (t, J = 9.4 Hz, 1H), 7.30 (d, J = 6.5 Hz, 1H), 7.25 (s, 1H), 7.23 (d, J = 7.9 Hz, 2H), 4.36 (q, J = 13.9, 7.0 Hz, 2H), 2.18 (s, 3H), 1.36 (t, J = 7.0 Hz, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 165.55, 159.84 (d, J = 242.3 Hz), 156.29, 147.81, 143.45, 143.01, 137.82, 135.57 (d, J = 7.6 Hz), 135.54, 131.43, 129.39, 128.30, 127.64, 127.50 (d, J = 9.9 Hz), 122.49, 116.69 (d, J = 24.6 Hz), 111.23 (d, J = 22.5 Hz), 61.32, 21.25, 14.66。

1.6 3-氯-2-苯氧基-6-氨基喹喔啉(P5a)的合成

3-氯-2-苯氧基-6-硝基喹喔啉6.00 mmol (P4a), 100 mL乙醇, 加入5%钯炭, N₂将空气排尽, 再缓慢通入H₂, 常温搅拌过夜, 反应液过滤, 滤液拌硅胶, 硅胶柱分离, 得亮黄色固体粉末, 收率41%。

同法合成P5b~P5f。

1.7 N-(2-苯氧基-3-氯喹喔啉-6基)丙烯酰胺(P6a)的合成

3-氯-2-苯氧基-6-氨基喹喔啉0.30 mmol (P5a), 10 mL二氯甲烷, 于0 ℃下加入丙烯酰氯0.45 mmol, 移置室温搅拌2 h, 反应液, 硅胶柱分离, 得浅黄色固体(P6a), 收率65%, mp: 154.8~156.2 ℃, ESI-MS (m/z): 324.10 [M-H]^－, ¹H NMR (400 MHz, DMSO-d₆) δ 11.11 (s, 1H), 8.99 (s, 1H), 8.40 (d, J = 9.2 Hz, 1H), 8.20 (d, J = 9.0 Hz, 1H), 8.01 (d, J = 7.6 Hz, 2H), 7.90~7.83 (m, 3H), 7.00 (dd, J = 16.9, 10.0 Hz, 1H), 6.86 (d, J = 16.5 Hz, 1H), 6.36 (d, J = 10.1 Hz, 1H); ¹³C NMR (101 MHz, DMSO-d₆) δ 164.13, 153.05, 152.38, 139.65, 139.58, 139.49, 135.56, 131.96, 130.31, 128.26, 127.69, 126.22, 124.41, 122.03, 115.60。

同法合成P6b~P6f。

N-(2-(3-甲氧基苯氧基)-3-氯喹喔啉-6基)丙烯酰胺(P6b), 浅黄色固体, 收率65%, mp: 151.4~152.7 ℃, ESI-MS (m/z): 354.00 [M-H]^－, ¹H NMR (400 MHz, DMSO-d₆) δ 10.59 (s, 1H), 8.49 (s, 1H), 7.91 (d, J = 9.1 Hz, 1H), 7.73 (d, J = 8.3 Hz, 1H), 7.41 (t, J = 7.5 Hz, 1H), 6.96 (d, J = 12.3 Hz, 1H), 6.92 (s, 2H), 6.50 (dd, J = 16.5, 10.1 Hz, 1H), 6.36 (d, J = 16.7 Hz, 1H), 5.86 (d, J = 9.8 Hz, 1H), 3.79 (s, 3H); ¹³CNMR (126 MHz, DMSO-d₆) δ 164.13, 160.91, 154.08, 152.32, 139.62, 139.59, 139.50, 135.62, 131.96, 130.70, 128.27, 127.74, 124.41, 115.58, 114.02, 112.01, 107.99, 55.89。

N-(2-(4-甲氧基苯氧基)-3-氯喹喔啉-6基)丙烯酰胺(P6c), 浅黄色固体, 收率67%, mp: 158.4~159.7 ℃, ESI-MS (m/z): 354.00 [M-H]^－, ¹H NMR (400 MHz, DMSO-d₆) δ 10.58 (s, 1H), 8.47 (s, 1H), 7.90 (d, J = 8.9 Hz, 1H), 7.69 (d, J = 9.2 Hz, 1H), 7.28 (d, J = 7.7 Hz, 2H), 7.05 (d, J = 7.9 Hz, 2H), 6.50 (dd, J = 16.8, 10.0 Hz, 1H), 6.35 (d, J = 16.9 Hz, 1H), 5.85 (d, J = 10.0 Hz, 1H), 3.82 (s, 3H); ¹³C NMR (126 MHz, DMSO-d₆) δ 164.11, 157.31, 152.73, 146.25, 139.59, 139.42, 139.34, 135.60, 131.97, 128.22, 127.60, 124.34, 123.05, 115.61, 115.19, 55.91。

N-(2-(3, 5-二甲氧基苯氧基)-3-氯喹喔啉-6基)丙烯酰胺(P6d), 浅黄色固体, 收率60%, mp: 147.9~149.5 ℃, ESI-MS (m/z): 384.00 [M-H]^－, ¹H NMR (400 MHz, DMSO-d₆) δ 10.60 (s, 1H), 8.49 (s, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.77 (d, J = 9.1 Hz, 1H), 6.56 (s, 2H), 6.55~6.48 (m, 1H), 6.47 (s, 1H), 6.36 (d, J = 16.8 Hz, 1H), 5.86 (d, J = 10.3 Hz, 1H), 3.77 (s, 6H); ¹³C NMR (126 MHz, DMSO-d₆) δ 164.14, 161.55, 154.74, 152.25, 139.61, 139.52, 135.69, 131.96, 131.90, 128.28, 127.82, 124.42, 115.57, 100.54, 98.30, 55.98。

N-(2-(3-氯苯氧基)-3-氯喹喔啉-6基)丙烯酰胺(P6e), 浅黄色固体, 收率70%, mp: 174.5~176.2 ℃, ESI-MS (m/z): 359.95 [M-H]^－, ¹H NMR (400 MHz, DMSO-d₆) δ 10.61 (s, 1H), 8.50 (s, 1H), 7.91 (d, J = 9.2 Hz, 1H), 7.75 (d, J = 9.2 Hz, 1H), 7.63~7.48 (m, 2H), 7.40 (dd, J = 18.3, 8.1 Hz, 2H), 6.50 (dd, J = 16.8, 10.1 Hz, 1H), 6.36 (d, J = 16.8 Hz, 1H), 5.86 (d, J = 10.3 Hz, 1H); ¹³C NMR (126 MHz, DMSO-d₆) δ 164.17, 153.78, 152.17, 139.78, 139.67, 139.54, 135.45, 134.10, 131.95, 131.73, 128.34, 127.81, 126.39, 124.49, 122.47, 121.05, 115.56。

N-(2-(3-氟苯氧基)-3-氯喹喔啉-6基)丙烯酰胺(P6f), 浅黄色固体, 收率68%, mp: 167.8~168.5 ℃, ESI-MS (m/z): 341.95 [M-H]^－, ¹H NMR (400 MHz, DMSO-d₆) δ 10.61 (s, 1H), 8.50 (s, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.75 (d, J = 9.0 Hz, 1H), 7.56 (dd, J = 15.3, 8.0 Hz, 1H), 7.36 (d, J = 9.8 Hz, 1H), 7.22 (dd, J = 19.1, 8.9 Hz, 2H), 6.50 (dd, J = 16.9, 10.1 Hz, 1H), 6.36 (d, J = 16.6 Hz, 1H), 5.86 (d, J = 10.1 Hz, 1H); ¹³C NMR (126 MHz, DMSO-d₆) δ 164.16, 162.92 (d, J = 244.9 Hz), 154.04 (d, J = 11.4 Hz), 152.08, 139.78, 139.69, 139.53, 135.47, 131.95, 131.55 (d, J = 9.6 Hz), 128.32, 127.82, 124.49, 118.27, 115.55, 113.20 (d, J = 20.9 Hz), 110.03 (d, J = 24.6 Hz)。

1.8 中间体核磁共振氢谱

2, 3-二氯喹喔啉(H3), ¹H NMR (400 MHz, DMSO-d₆) δ 8.13~8.05 (m, 2H), 7.99~7.92 (m, 2H)。

2-氯-3-苯氧基喹喔啉(H4a), ¹H NMR (400 MHz, DMSO-d₆) δ 8.06~7.98 (m, 1H), 7.80~7.66 (m, 3H), 7.53 (t, J = 7.9 Hz, 2H), 7.36 (dd, J = 15.0, 7.7 Hz, 3H)。

2-肼基-3-苯氧基喹喔啉(H5a), ¹H NMR (400 MHz, DMSO-d₆) δ 8.95 (s, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.50 (t, J = 7.9 Hz, 2H), 7.46~7.39 (m, 2H), 7.32 (dd, J = 13.1, 7.6 Hz, 3H), 7.24 (t, J = 7.1 Hz, 1H), 4.59 (s, 2H)。

2-氯-3-(3-甲氧基苯氧基)喹喔啉(H4b), ¹H NMR (400 MHz, DMSO-d₆) δ 8.14~7.88 (m, 1H), 7.80~7.71 (m, 3H), 7.41 (t, J = 8.2 Hz, 1H), 7.00 (t, J = 2.2 Hz, 1H), 6.93 (td, J = 8.6, 2.2 Hz, 2H), 3.78 (s, 3H)。

2-肼基-3-(3-甲氧基苯氧基)喹喔啉(H5b), ¹H NMR (400 MHz, DMSO-d₆) δ 8.92 (s, 1H), 7.60 (d, J = 7.6 Hz, 1H), 7.45~7.40 (m, 2H), 7.38 (t, J = 8.2 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 6.92 (t, J = 5.2 Hz, 2H), 6.90~6.85 (m, 1H), 4.57 (s, 2H), 3.78 (s, 3H)。

2-氯-3-(4-甲氧基苯氧基)喹喔啉(H4c), ¹H NMR (400 MHz, DMSO-d₆) δ 8.01 (d, J = 7.5 Hz, 1H), 7.74 (dt, J = 14.6, 7.4 Hz, 3H), 7.29 (d, J = 9.0 Hz, 2H), 7.05 (d, J = 9.0 Hz, 2H), 3.81 (s, 3H)。

2-肼基-3-(4-甲氧基苯氧基)喹喔啉(H5c), ¹H NMR (400 MHz, DMSO-d₆) δ 8.91 (s, 1H), 7.59 (d, J = 7.9 Hz, 1H), 7.41 (t, J = 8.5 Hz, 2H), 7.24 (dd, J = 12.7, 6.3 Hz, 3H), 7.03 (d, J = 9.0 Hz, 2H), 4.57 (s, 2H), 3.80 (s, 3H)。

2-氯-3-(3, 5-二甲氧基苯氧基)喹喔啉(H4d), ¹H NMR (400 MHz, DMSO-d₆) δ 8.02 (d, J = 6.9 Hz, 1H), 7.83~7.68 (m, 3H), 6.58 (d, J = 2.0 Hz, 2H), 6.48 (s, 1H), 3.76 (s, 6H)。

2-肼基-3-(3, 5-二甲氧基苯氧基)喹喔啉(H5d), ¹H NMR (400 MHz, DMSO-d₆) δ 8.91 (s, 1H), 7.60 (d, J = 8.1 Hz, 1H), 7.45 (dd, J = 17.1, 7.6 Hz, 2H), 7.26 (dd, J = 11.0, 4.1 Hz, 1H), 6.55 (d, J = 2.2 Hz, 2H), 6.45 (t, J = 2.2 Hz, 1H), 4.57 (s, 2H), 3.76 (s, 6H)。

2-氯-3-(3-氯苯氧基)喹喔啉(H4e), ¹H NMR (400 MHz, DMSO-d₆) δ 8.03 (d, J = 8.3 Hz, 1H), 7.76 (d, J = 6.1 Hz, 2H), 7.59 (d, J = 2.1 Hz, 1H), 7.55 (d, J = 8.1 Hz, 1H), 7.43 (d, J = 8.1 Hz, 1H), 7.39 (dd, J = 8.2, 2.2 Hz, 1H)。

2-肼基-3-(3-氯苯氧基)喹喔啉(H5e), ¹H NMR (400 MHz, DMSO-d₆) δ 8.97 (s, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.53 (t, J = 8.2 Hz, 2H), 7.45 (d, J = 7.9 Hz, 2H), 7.38 (t, J = 9.2 Hz, 2H), 7.25 (t, J = 7.7 Hz, 1H), 4.60 (s, 2H)。

2-氯-3-(3-氯苯氧基)喹喔啉(H4f), ¹H NMR (400 MHz, DMSO-d₆) δ 8.03 (d, J = 7.7 Hz, 1H), 7.80~7.71 (m, 3H), 7.60~7.57 (m, 2H), 7.47~7.41 (m, 2H)。

2-肼基-3-(4-氯苯氧基)喹喔啉(H5f), ¹H NMR (400 MHz, DMSO-d₆) δ 8.98 (s, 1H), 7.61 (d, J = 8.6 Hz, 1H), 7.58~7.53 (m, 2H), 7.43 (d, J = 7.6 Hz, 2H), 7.42~7.39 (m, 2H), 7.25 (dd, J = 11.1, 4.0 Hz, 1H), 4.59 (s, 2H)。

2-氯-3-(3-氟苯氧基)喹喔啉(H4g), ¹H NMR (400 MHz, DMSO-d₆) δ 8.04 (d, J = 8.5 Hz, 1H), 7.78 (s, 1H), 7.70 (dd, J = 6.2, 3.5 Hz, 1H), 7.62~7.54 (m, 2H), 7.39 (dd, J = 8.9, 6.9 Hz, 1H), 7.29 (d, J = 10.4 Hz, 1H), 7.21 (t, J = 8.5 Hz, 1H)。

2-肼基-3-(3-氟苯氧基)喹喔啉(H5g), ¹H NMR (400 MHz, DMSO-d₆) δ 8.97 (s, 1H), 7.61 (d, J = 7.4 Hz, 1H), 7.54 (dd, J = 15.1, 8.1 Hz, 1H), 7.48~7.42 (m, 2H), 7.34 (d, J = 10.1 Hz, 1H), 7.25 (dd, J = 13.3, 7.0 Hz, 2H), 7.18 (t, J = 8.4 Hz, 1H), 4.59 (s, 2H)。

4-((3-氯喹喔啉-2-基)氧基)苯甲酸乙酯(H4h), ¹H NMR (400 MHz, DMSO-d₆) δ 8.13~8.09 (m, 2H), 8.04 (dd, J = 6.7, 2.8 Hz, 1H), 7.82~7.72 (m, 3H), 7.57~7.52 (m, 2H), 4.36 (q, J = 7.1 Hz, 2H), 1.35 (t, J = 7.1 Hz, 3H)。

4-((3-肼基喹喔啉-2-基)氧基)苯甲酸乙酯(H5h), ¹H NMR (400 MHz, DMSO-d₆) δ 9.02 (s, 1H), 8.08 (d, J = 8.6 Hz, 2H), 7.62 (d, J = 8.2 Hz, 1H), 7.51 (d, J = 8.6 Hz, 2H), 7.48~7.41 (m, 2H), 7.27 (d, J = 7.0 Hz, 1H), 4.60 (s, 2H), 4.34 (q, J = 7.1 Hz, 2H), 1.34 (t, J = 7.1 Hz, 3H)。

2, 3-二氯-6-氟喹喔啉(F3), ¹H NMR (400 MHz, DMSO-d₆) δ 8.19 (dd, J = 7.7, 6.1 Hz, 1H), 7.96 (d, J = 9.2 Hz, 1H), 7.90 (t, J = 8.9 Hz, 1H)。

3-氯-6-氟-2-苯氧基喹喔啉(F4a), ¹H NMR (400 MHz, DMSO-d₆) δ 8.10 (t, J = 7.0 Hz, 1H), 7.66 (t, J = 8.8 Hz, 2H), 7.54 (dd, J = 14.6, 8.0 Hz, 3H), 7.39 (d, J = 6.9 Hz, 2H), 7.34 (s, 1H)。

3-肼基-6-氟-2-苯氧基喹喔啉(F5a), ¹H NMR (400 MHz, DMSO-d₆) δ 8.95 (s, 1H), 7.66~7.60 (m, 1H), 7.50 (t, J = 7.6 Hz, 2H), 7.42~7.26 (m, 4H), 7.22 (d, J = 10.2 Hz, 1H), 4.57 (s, 2H)。

3-氯-2-(3-甲氧基苯氧基)-6-氟喹喔啉(F4b), ¹H NMR (400 MHz, DMSO-d₆) δ 8.10 (dd, J = 9.2, 5.9 Hz, 1H), 7.66 (td, J = 8.8, 2.8 Hz, 1H), 7.60 (dd, J = 9.8, 2.7 Hz, 1H), 7.42 (t, J = 8.2 Hz, 1H), 7.00 (t, J = 2.3 Hz, 1H), 6.93 (td, J = 7.8, 2.3 Hz, 2H), 3.78 (s, 3H)。

3-肼基-2-(3-甲氧基苯氧基)-6-氟喹喔啉(F5b), ¹H NMR (400 MHz, DMSO-d₆) δ 8.93 (s, 1H), 7.62 (dd, J = 9.0, 5.9 Hz, 1H), 7.39 (t, J = 8.2 Hz, 1H), 7.32 (td, J = 8.8, 2.9 Hz, 1H), 7.25 (dd, J = 9.8, 2.9 Hz, 1H), 6.96~6.87 (m, 3H), 4.56 (s, 2H), 3.78 (s, 3H)。

3-氯-2-(4-甲氧基苯氧基)-6-氟喹喔啉(F4c), ¹H NMR (400 MHz, DMSO-d₆) δ 8.21 (dd, J = 9.3, 5.9 Hz, 1H), 8.03~7.96 (m, 1H), 7.69~7.61 (m, 1H), 7.31 (d, J = 8.2 Hz, 2H), 7.24 (d, J = 8.5 Hz, 2H), 2.37 (s, 3H)。

3-肼基-2-(4-甲氧基苯氧基)-6-氟喹喔啉(F5c), ¹H NMR (400 MHz, DMSO-d₆) δ 8.94 (s, 1H), 7.61 (dd, J = 9.0, 5.9 Hz, 1H), 7.35~7.30 (m, 1H), 7.30~7.26 (m, 2H), 7.24~7.15 (m, 3H), 4.55 (s, 2H), 2.36 (s, 3H)。

3-氯-2-(3, 5-二甲氧基苯氧基)-6-氟喹喔啉(F4d), ¹H NMR (400 MHz, DMSO-d₆) δ 8.20~7.99 (m, 1H), 7.73~7.53 (m, 1H), 6.57 (s, 2H), 6.47 (s, 1H), 3.75 (s, 6H)。

3-肼基-2-(3, 5-二甲氧基苯氧基)-6-氟喹喔啉(F5d), ¹H NMR (400 MHz, DMSO-d₆) δ 8.88 (s, 1H), 7.69~7.55 (m, 1H), 7.30 (dd, J = 16.4, 9.0 Hz, 2H), 6.55 (s, 2H), 6.45 (s, 1H), 4.53 (s, 2H), 3.75 (s, 6H)。

3-氯-2-(3-氯苯氧基)-6-氟喹喔啉(F4e), ¹H NMR (400 MHz, DMSO-d₆) δ 8.11 (dd, J = 9.1, 5.8 Hz, 1H), 7.72~7.65 (m, 1H), 7.62 (dd, J = 9.7, 2.7 Hz, 1H), 7.58 (t, J = 2.0 Hz, 1H), 7.55 (d, J = 8.1 Hz, 1H), 7.47~7.42 (m, 1H), 7.39 (dd, J = 7.8, 1.8 Hz, 1H)。

3-肼基-2-(3-氯苯氧基)-6-氟喹喔啉(F5e), ¹H NMR (400 MHz, DMSO-d₆) δ 8.96 (s, 1H), 7.67~7.61 (m, 1H), 7.54 (t, J = 8.1 Hz, 2H), 7.43~7.36 (m, 2H), 7.33 (d, J = 9.4 Hz, 1H), 7.27 (d, J = 9.7 Hz, 1H), 4.62 (s, 2H)。

3-氯-2-(3-氯苯氧基)-6-氟喹喔啉(F4f), ¹H NMR (400 MHz, DMSO-d₆) δ 8.10 (dd, J = 9.2, 5.8 Hz, 1H), 7.67 (td, J = 8.8, 2.8 Hz, 1H), 7.60 (dd, J = 6.6, 4.5 Hz, 3H), 7.47~7.40 (m, 2H)。

3-肼基-2-(3-氯苯氧基)-6-氟喹喔啉(F5f), ¹H NMR (400 MHz, DMSO-d₆) δ 8.98 (s, 1H), 7.67~7.59 (m, 1H), 7.56 (d, J = 8.7 Hz, 2H), 7.41 (d, J = 8.8 Hz, 2H), 7.37~7.30 (m, 1H), 7.25 (d, J = 10.2 Hz, 1H), 4.65 (s, 2H)。

3-氯-2-(3-氟苯氧基)-6-氟喹喔啉(F4g), ¹H NMR (400 MHz, DMSO-d₆) δ 8.15~8.08 (m, 1H), 7.69 (t, J = 8.9 Hz, 1H), 7.62 (d, J = 9.8 Hz, 1H), 7.60~7.54 (m, 1H), 7.38 (d, J = 8.1 Hz, 1H), 7.29~7.21 (m, 2H)。

3-肼基-2-(3-氟苯氧基)-6-氟喹喔啉(F5g), ¹H NMR (400 MHz, DMSO-d₆) δ 8.97 (s, 1H), 7.70~7.58 (m, 1H), 7.54 (t, J = 8.1 Hz, 2H), 7.44~7.36 (m, 2H), 7.36~7.31 (m, 1H), 7.27 (d, J = 10.8 Hz, 1H), 4.65 (s, 2H)。

4-((3-氯-6-氟喹喔啉-2-基)氧基)苯甲酸乙酯(F4h), ¹H NMR (400 MHz, DMSO-d₆) δ 8.12 (t, J = 7.5 Hz, 3H), 7.69 (t, J = 8.8 Hz, 1H), 7.56 (t, J = 10.1 Hz, 3H), 4.36 (q, J = 6.9 Hz, 2H), 1.36 (t, J = 7.0 Hz, 3H)。

4-[(3-肼基-6-氟喹喔啉-2-基)氧基]苯甲酸乙酯(F5h), ¹H NMR (400 MHz, DMSO-d₆) δ 9.00 (s, 1H), 8.17~8.04 (m, 2H), 7.73~7.66 (m, 1H), 7.66~7.58 (m, 1H), 7.58~7.49 (m, 2H), 7.40~7.21 (m, 1H), 4.59 (s, 2H), 4.37 (q, 2H), 1.36 (t, J = 7.1 Hz, 3H)。

2, 3-二氯-6-硝基喹喔啉(P3), ¹H NMR (400 MHz, DMSO-d₆) δ 8.95~8.86 (m, 1H), 8.62 (dd, J = 9.2, 0.7 Hz, 1H), 8.33 (d, J = 9.2 Hz, 1H)。

3-氯-6-硝基-2-苯氧基喹喔啉(P4a), ¹H NMR (400 MHz, DMSO-d₆) δ 8.83 (d, J = 2.4 Hz, 1H), 8.44 (dd, J = 9.2, 2.6 Hz, 1H), 7.90 (d, J = 9.2 Hz, 1H), 7.59~7.50 (m, 2H), 7.41 (d, J = 7.7 Hz, 2H), 7.37 (d, J = 7.4 Hz, 1H)。

3-氯-2-(3-甲氧基苯氧基)-6-硝基喹喔啉(P4b), ¹H NMR (400 MHz, DMSO-d₆) δ 8.84 (d, J = 2.5 Hz, 1H), 8.45 (dd, J = 9.2, 2.6 Hz, 1H), 7.94 (d, J = 9.2 Hz, 1H), 7.44 (t, J = 8.2 Hz, 1H), 7.02 (t, J = 2.3 Hz, 1H), 7.00~6.93 (m, 2H), 3.78 (s, 3H)。

3-氯-2-(4-甲氧基苯氧基)-6-硝基喹喔啉(P4c), ¹H NMR (400 MHz, DMSO-d₆) δ 8.82 (s, 1H), 8.47~8.41 (m, 1H), 7.90 (d, J = 9.2 Hz, 1H), 7.32 (d, J = 9.0 Hz, 2H), 7.07 (d, J = 9.0 Hz, 2H), 3.82 (s, 3H)。

3-氯-2-(3, 5-二甲氧基苯氧基)-6-硝基喹喔啉(P4d), ¹H NMR (400 MHz, DMSO-d₆) δ 8.84 (d, J = 2.5 Hz, 1H), 8.45 (dd, J = 9.2, 2.6 Hz, 1H), 7.98 (d, J = 9.2 Hz, 1H), 6.62 (s, 1H), 6.61 (s, 1H), 6.51 (t, J = 2.2 Hz, 1H), 3.77 (s, 6H)。

3-氯-2-(3-氯苯氧基)-6-硝基喹喔啉(P4e), ¹H NMR (400 MHz, DMSO-d₆) δ 8.85 (d, J = 2.5 Hz, 1H), 8.46 (dd, J = 9.2, 2.6 Hz, 1H), 7.96 (d, J = 9.2 Hz, 1H), 7.62~7.60 (m, 1H), 7.58 (d, J = 8.1 Hz, 1H), 7.47 (ddd, J = 8.1, 2.0, 0.9 Hz, 1H), 7.42 (ddd, J = 8.2, 2.2, 0.9 Hz, 1H)。

3-氯-2-(3-氟苯氧基)-6-硝基喹喔啉(P4f), ¹H NMR (400 MHz, DMSO-d₆) δ 8.85 (d, J = 2.5 Hz, 1H), 8.46 (dd, J = 9.2, 2.6 Hz, 1H), 7.95 (d, J = 9.2 Hz, 1H), 7.60 (td, J = 8.3, 6.8 Hz, 1H), 7.40 (dt, J = 9.9, 2.3 Hz, 1H), 7.32~7.22 (m, 2H)。

3-氯-2-苯氧基6-氨基喹喔啉(P5a), ¹H NMR (400 MHz, DMSO-d₆) δ 7.55 (d, J = 2.5 Hz, 1H), 7.42 (dd, J = 6.2, 2.6 Hz, 1H), 6.80 (d, J = 7.2 Hz, 1H), 6.64~6.60 (m, 2H), 6.41 (d, J = 7.2 Hz, 2H), 6.37 (d, J = 6.4 Hz, 1H), 5.94 (s, 2H)。

3-氯-2-(3-甲氧基苯氧基)-6-氨基喹喔啉(P5b), ¹H NMR (400 MHz, DMSO-d₆) δ 7.46 (d, J = 9.0 Hz, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.16 (d, J = 8.8 Hz, 1H), 6.90 (d, J = 12.4 Hz, 2H), 6.82 (d, J = 8.5 Hz, 2H), 5.96 (s, 2H), 3.78 (s, 3H)。

3-氯-2-(4-甲氧基苯氧基)-6-氨基喹喔啉(P5c), ¹H NMR (400 MHz, DMSO-d₆) δ 7.41 (d, J = 9.1 Hz, 1H), 7.21 (d, J = 7.8 Hz, 2H), 7.13 (d, J = 9.2 Hz, 1H), 7.01 (d, J = 7.7 Hz, 2H), 6.90 (s, 1H), 5.91 (s, 2H), 3.80 (s, 3H)。

3-氯-2-(3, 5-二甲氧基苯氧基)-6-氨基喹喔啉(P5d), ¹H NMR (400 MHz, DMSO-d₆) δ 7.48 (d, J = 8.7 Hz, 1H), 7.15 (d, J = 9.0 Hz, 1H), 6.89 (s, 1H), 6.44 (s, 2H), 6.41 (s, 1H), 5.95 (s, 2H), 3.74 (s, 6H)。

3-氯-2-(3-氯苯氧基)-6-氨基喹喔啉(P5e), ¹H NMR (400 MHz, DMSO-d₆) δ 7.51 (d, J = 8.3 Hz, 1H), 7.47 (d, J = 7.2 Hz, 2H), 7.36 (d, J = 8.2 Hz, 1H), 7.28 (d, J = 8.3 Hz, 1H), 7.17 (d, J = 9.2 Hz, 1H), 6.91 (s, 1H), 5.99 (s, 2H)。

3-氯-2-(3-氟苯氧基)-6-氨基喹喔啉(P5f), ¹H NMR (400 MHz, DMSO-d₆) δ 7.55~7.49 (m, 1H), 7.48 (d, J = 9.1 Hz, 1H), 7.26 (d, J = 10.3 Hz, 1H), 7.16 (t, J = 9.0 Hz, 3H), 6.91 (s, 1H), 6.00 (s, 2H)。

2 生物活性测试

用LY294002作为阳性对照化合物, 采用MTT法, 以人非小细胞肺癌A549、人乳腺癌细胞MCF-7、人结肠癌细胞HCT-116和人肝癌细胞HepG2 4种癌细胞为筛选对象, 分别对22个目标化合物进行体外抗肿瘤活性测试。于96孔板中接种处于对数期的癌细胞每孔5×10³个, 培养24 h后加入待测化合物。继续培养48 h后加入MTT溶液20 μL, 培养4 h后弃上清液, 加入DMSO 200 mL, 振匀, 酶标仪570 nm波长测OD值, 重复3次, 取平均值, 计算得IC₅₀。抑制率(%) = ((对照组OD值-给药组OD值)/对照组OD值)×100%。

基金

收起

福建省科技创新联合资金资助项目(2016Y9052)
福建省高校杰出青年科研人才培育计划(2017B021)
福建医科大学引进高层次创业创新人才计划(XRCGZX2017010)
福建省自然科学基金项目(2018J01846)
福建省自然科学基金青年创新项目(2019J05073)

参考文献

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https://doi.org/10.1016/j.ejmech.2008.07.025

2020年第55卷第1期

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doi: 10.16438/j.0513-4870.2019-0679

接收时间：2019-08-26
首发时间：2026-01-20
出版时间：2020-01-12

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收稿日期：2019-08-26
修回日期：2019-10-21

基金

福建省科技创新联合资金资助项目(2016Y9052)

福建省高校杰出青年科研人才培育计划(2017B021)

福建医科大学引进高层次创业创新人才计划(XRCGZX2017010)

福建省自然科学基金项目(2018J01846)

福建省自然科学基金青年创新项目(2019J05073)

作者信息

1.福建省儿童医院(福建省妇幼保健院), 福建福州 350001

2.福建医科大学药学院, 福建省药物靶点发现与结构功能研究重点实验室, 福建福州 350122

通讯作者:

*李柱来, E-mail:lizhulai@126.com;

许秀枝, Tel:86-591-22862016, E-mail:xiuzhi_xu@126.com

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https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2019-0679

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Compd.	R₁	R₂	IC₅₀/μmol·L^-1
Compd.	R₁	R₂	A549	MCF-7	HCT-116	HepG2
H6a	H	3-H	61.92	200 >	59.58	78.82
H6b	H	3-Methoxy	157.3	200 >	35.59	142
H6c	H	4-Methoxy	282.6	200 >	42.65	66.44
H6d	H	3, 5-Dimethoxy	200 >	200 >	200 >	200 >
H6e	H	3-Chloro	41.77	82.83	42.65	40.68
H6f	H	4-Chloro	29.09	86.67	33.38	41.63
H6g	H	3-Fluoro	52.53	83.85	27.81	49.51
H6h	H	4-COOC₂H₅	200 >	147.4	10.88	17.61
F6a	Fluoro	3-H	143.4	81.22	58.78	144.9
F6b	Fluoro	3-Methoxy	200 >	200 >	118.3	200 >
F6c	Fluoro	4-Methoxy	200 >	120.2	78.57	141.9
F6d	Fluoro	3, 5-Dimethoxy	126.6	105.8	200 >	200 >
F6e	Fluoro	3-Chloro	152.9	200 >	142.3	200 >
F6f	Fluoro	4-Chloro	137.4	200 >	79.19	183.1
F6g	Fluoro	3-Fluoro	126	200	200 >	200 >
F6h	Fluoro	4-COOC₂H₅	200 >	200 >	200 >	200 >
LY294002	-	-	23.60	20.61	11.80	11.41