细胞分裂周期25磷酸酯酶B抑制剂双1, 2, 4-三唑多杂环分子的设计合成

引用本文: 张成路, 王静, 李益政, 李传银, 孙晓娜. 细胞分裂周期25磷酸酯酶B抑制剂双1, 2, 4-三唑多杂环分子的设计合成[J]. 应用化学, 2018, 35(2): 154-164. doi: 10.11944/j.issn.1000-0518.2018.02.170004 shu

Citation: ZHANG Chenglu, WANG Jing, LI Yizheng, LI Chuanyin, SUN Xiaona. Synthesis and Design of Multi-Heterocyclic Molecules with Duoble 1, 2, 4-Triazole Motifs as Inhibitors of Cell Division Cycle Protein 25B Phosphatases[J]. Chinese Journal of Applied Chemistry, 2018, 35(2): 154-164. doi: 10.11944/j.issn.1000-0518.2018.02.170004 shu

细胞分裂周期25磷酸酯酶B抑制剂双1, 2, 4-三唑多杂环分子的设计合成

辽宁师范大学化学化工学院辽宁大连 116029

通讯作者: 张成路, 教授, Tel:0411-84203175, E-mail:zhangchenglu@lnnu.edu.cn, 研究方向:药物合成

基金项目:
辽宁省教育厅科学技术项目资助（2009A426）

摘要: 以1，2，4-三唑为核心结构的杂环分子，因其优良而广泛的生物活性，已成为药物筛选中重要的研究对象。本文为了研究其构效关系，针对1，2，4-三唑结构中3个位点分别进行不同的结构修饰，并将其与哌嗪和均三嗪分别偶联，首次合成了两类共28个双三唑多杂环目标分子[TM1~TM3（a~f）]和[TM4~TM5（a~e）]。为了比较三唑中4-氨基对生物活性的影响，还与化合物6-吗啉-2，4-（5-硫醚基-4-氨基-3-正戊基1，2，4-三唑）-1，3，5-三嗪（C）和6-吡咯-2，4-（5-巯基-4-氨基-3-正戊基-1，2，4-三唑）-1，3，5-三嗪（D）进行了比较。通过傅里叶变换红外光谱仪（IR）、核磁共振谱（¹H NMR）和高分辨率质谱仪（HRMS）等技术手段对28个目标分子进行了结构表征，研究了其对细胞分裂周期25磷酸酯酶B（Cdc25B）的抑制活性。结果表明，21个目标分子表现出优良的抑制活性，9个目标分子的抑制活性优于阳性参照物，有望成为抗肿瘤药物先导物。

关键词:

English

Synthesis and Design of Multi-Heterocyclic Molecules with Duoble 1, 2, 4-Triazole Motifs as Inhibitors of Cell Division Cycle Protein 25B Phosphatases

College of Chemistry and Chemical Engineering, Liaoning Normal University, Dalian, Liaoning 116029, China

Corresponding author: ZHANG Chenglu, professor; Tel:0411-84203175; E-mail:zhangchenglu@lnnu.edu.cn; Research interests:drug synthesis

Received Date: 03 January 2017
Accepted Date: 14 April 2017
Revised Date: 07 March 2017
Available Online: 10 February 2018
Fund Project:
Supported by the Technology Research Program of Liaoning Provincial Department of Education(No.2009A426)

Abstract: Heterocyclic molecules containing 1, 2, 4-triazole motif are important for drug screening due to their better broad spectrum of bioactivities. For structure-activity relationship study, twenty-eight novel multi-heterocyclic molecules[TM1~TM3(a~f)] and[TM4~TM5(a~e)] containing duo 1, 2, 4-triazole motifs were synthesized by the conjugation with piperazine and s-triazine, respectively. The structures of target molecules were confirmed via infrared(IR), proton nuclear magnetic resonance(¹H NMR) and high resolution mass spectrometry(HRMS). In order to investigate the effect of 4-amino substituent on 1, 2, 4-triazole for bioactivities, the comparison between compounds 6-morpholine-2, 4-(5-thioether-4-amino-3-n-pentyl-1, 2, 4-triazole)-1, 3, 5-triazine(C), 6-pyrrole-2, 4-(5-mercapto-4-amino-3-n-pentyl-1, 2, 4-triazole)-1, 3, 5-triazine(D) and target molecules were also performed. The inhibitory activities against cell division cycle protein 25B(Cdc25B) phosphatases of synthesized molecules were evaluated. Twenty one molecules exhibit good activities, and nine among them are more potent than the positive control Na₃VO₄.

Key words:

1, 2, 4-triazole
/ piperazine
/ s-triazine
/ cell division cycle protein 25B inhibitor

癌症被认为是最具破坏性的疾病之一，因此新型抗癌药物的研制已成为化学、药学领域研究的重点^[1]。细胞分裂周期25(cell division cycle 25, 缩写Cdc25)磷酸酯酶B(Cdc25B)是双特异性蛋白磷酸酯酶，其活性调节主要通过磷酸化和去磷酸化来实现。Cdc25B是一种潜在的癌基因，在人类多种癌细胞系中过度表达，具有促进细胞有丝分裂的作用，对G2-M期的待修复检验点起着重要的作用，因此寻求性能优异的Cdc25B抑制剂将十分重要。

取代杂环分子的结构具有高度多样性，存在于许多天然产物和临床药物中，已被证明具有显著药理活性如抗菌、止痛、抗炎、抗癌和抗氧化特性等，如泊沙康唑和雷夫康唑^[2-5]。在众多杂环组块中，1, 2, 4-三唑衍生物因其分子结构中存在＝N—N＝C—N活性基团，能通过氢键等与生物体内的多种酶等结合从而产生不同的药效作用^[6-8]，如抗菌^[9-11]、抗病毒^[12]和镇痛^[13-14]等。尤其是在许多已应用于临床的药物分子中结构中含有1, 2, 4-三唑活性组块，如低毒杀菌剂三唑酮，可用于治疗深部真菌感染的伊曲康唑和用于治疗肺癌的氯唑等。均三嗪衍生物也具有抗菌、抗癌、抗疟药和抗病毒等广泛的活性，成为药物筛选中重要的研究对象^[15-16]。如在临床上用于治疗肺癌、乳腺癌的六甲蜜胺等，表现出优良的MT-4细胞的抗HIV活性，优于奈韦拉平、地拉韦啶和齐多夫定等，有望成为抗人类免疫缺陷病毒1型(HIV-1)的先导药物。

哌嗪活性组块也存在于许多分子中，具有抗微生物^[17]、抗癌^[18]、除草和杀虫等活性^[19]。在临床药物伊曲康唑中哌嗪与1, 2, 4-三唑酮组合发挥了优良的药用价值。多种哌嗪衍生物可作为乳腺癌、前列腺癌、白血病和肝脏癌细胞的抑制剂。因此，哌嗪常被引入到多种杂环衍生结构中，以增强分子的生物活性。

为了筛选优良的Cdc25B抑制剂，探究不同药效基团在分子中相同位点和相同基团在不同位点对分子生物活性的影响，本文以1, 2, 4-三唑为模板，分别在其3号位引入对氯苯基、对甲氧苯基和正戊基，然后借助4号位分别引入氨基和构筑Schiff碱，并将两个1, 2, 4-三唑衍生的活性组块分别与1, 3, 5-三嗪和哌嗪对接，设计了两类共28个杂环分子，期望从不同角度通过研究其构效关系，筛选出活性优良的Cdc25B抑制剂，进而获得抗肿瘤药物先导化合物。目标分子合成路线如Scheme 1所示。

Scheme1. Synthetic routes of target molecules

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1 实验部分

1.1 仪器和试剂

BRUKER Avance-500MHz型核磁共振波谱仪(NMR, 瑞士Bruker公司)；Agilent 6224型高分辨质谱仪(HRMS, 美国Agilent公司)；TENSOR 27型傅里叶变换红外光谱仪(FT-IR, 德国Bruker AXS公司)；X-5型数字显微熔点测定仪(北京泰克仪器有限公司)；WFH-203B型三用紫外分析仪(上海精科实业有限公司)。

无水乙醇、甲醛、苯甲醛及哌嗪购自国药集团化学试剂公司；浓硫酸、水合肼和甲苯购自科密欧化学试剂厂；吡咯烷、吗啡啉购自阿拉丁化学试剂公司，所用试剂均为分析纯。

1.2 实验方法

1.2.1 化合物1和化合物2的合成

化合物1根据文献[20]的方法合成，产物为白色固体, 收率75.8%，mp 190.7~192.1 ℃，¹H NMR(CDCl₃), δ:12.59(s, 1H, SH), 7.94(d, J=8.80 Hz, 2H, PhH), 7.02(d, J=8.80 Hz, 2H, PhH), 5.78(s, 2H, NH₂)。

化合物2根据文献[21]的方法合成，产物为白色固体, 收率74.3%，mp 204.2~206.7 ℃，¹H NMR(DMSO-d₆), δ:13.67(s, 1H, SH), 7.87(d, J=8.80 Hz, 2H, PhH), 6.89(d, J=8.80 Hz, 2H, PhH), 5.89(s, 2H, NH₂), 3.87(s, 3H, OCH₃)。

1.2.2 化合物3的合成

N₂气氛围下，量取4 mmol脂肪酸加入100 mL三口瓶中，50 ℃搅拌20 min。加入1.6 mmol化合物硫代对称二氨基硫脲，升温至150 ℃，剧烈搅拌5 h，冷却为白色块状固体，用乙醇多次重结晶，滤出白色固体，用适量石油醚剧烈搅拌2 h，过滤，干燥等到白色固体产品^[22]。收率65.7%, mp 120.3~121.7 ℃，¹H NMR(CDCl₃), δ:13.45(s, 1H, SH), 5.89(s, 2H, NH₂), 2.87(t, J=8.85 Hz, 2H, CH₂), 1.81(t, J=7.43 Hz, 2H, CH₂), 1.31(t, J=7.03 Hz, 4H, CH₂), 0.89(t, J=7.00 Hz, 3H, CH₃)。

1.2.3 中间体4a~4f、5a~5f和6a~6f的合成

于50 mL的圆底烧瓶中，分别加入化合物1、2、3(0.5 mmol)和乙醇5 mL，加热至回流后，加入芳香醛ArCHO(0.6 mmol)，TLC检测反应完全后，降至室温，加适量水有固体析出，过滤后将粗产品用乙醇重结晶，得目标产品。

1.2.4 化合物A和B的合成

化合物A和B根据文献[23]的方法合成，产物均为白色固体, 化合物A收率73.9%, mp 172.7~174.5 ℃，¹H NMR(CDCl₃), δ:3.84(t, J=5.60 Hz, 4H, CH₂OCH₂), 3.62(t, J=4.89 Hz, 4H, CH₂NCH₂)。

化合物B 收率74.2%, mp 122.0~124.6 ℃，¹H NMR(CDCl₃), δ:3.74(t, J=4.90 Hz, 4H, CH₂N), 1.63~1.60(m, 4H, CH₂)。所得结果均与文献值相符。

1.2.5 化合物C和D的合成方法

化合物C和D根据文献[24]的方法合成，产物均为白色固体, 化合物C收率81%, mp 251.2~252.5 ℃，¹H NMR(DMSO-d₆), δ:6.06(s, 2H, NH₂), 3.48(t, J＝4.70 Hz, 2H, morpholine), 3.36(t, J=4.70 Hz, 2H, morpholine), 2.89(t, J＝7.20 Hz, 2H, CH₂), 1.80(t, J＝7.20 Hz, 2H, CH₂), 1.36(t, J＝7.00 Hz, 4H, CH₂), 0.91(t, J＝7.00 Hz, 3H, CH₃)。所得结果与文献值相符。

化合物D 收率82%, mp 251.8~252.4 ℃，¹H NMR(DMSO-d₆), δ:6.19(s, 2H, NH₂), 3.22(d, J＝4.90 Hz, 2H, pyrrolidine), 2.80(t, J＝7.20 Hz, 2H, CH₂), 1.88(t, J＝7.20 Hz, 2H, CH₂), 1.38~1.35(m, 2H, CH₂), 1.33(t, J＝7.00 Hz, 4H, CH₂), 0.84(t, J＝7.00 Hz, 3H, CH₃), 2.89(t, J=7.3 Hz, 2H, CH₂), 1.72~1.69(m, 2H, CH₂), 1.39~1.32(m, 2H, CH₂), 1.21~1.15(m, 14H, CH₂), 0.79(t, J=6.8 Hz, 3H, CH₃)。所得结果与文献值相符。

1.2.6 目标分子TM1a~1f、TM2a~2f和TM3a~3f的合成

于50 mL的圆底烧瓶中，加入1 mol的Schiff碱(4a~4f、5a~5f)和8 mL乙醇。加热至回流后，加入6 mol的37%甲醛。搅拌10 min后，加入3 mol哌嗪。原料完全反应后，降至室温，加入少量水至固体不再析出。静置过夜，抽滤。粗产品用乙醇重结晶后得目标产品。

1.2.7 目标分子TM4a~4e和TM5a~5e的合成

取5 mmol Schiff碱于三颈瓶中，加入10 mL的THF，搅拌溶解，加入1 mmol的KOH，搅拌温度升至66 ℃，加入2.5 mmol均三嗪单取代物(A或B)，反应时间约为24 h。反应停止后，静置冷却至室温，过滤，将黄色滤液蒸去溶剂，得粗产品，然后用乙醚充分洗涤后得目标产品。

1.3 化合物的表征

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-氯苯基亚甲基氨基)-3-(4-氯苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM1a) 白色固体, 收率70%, mp 252.4~253.1 ℃, IR(KBr), σ/cm^-1:3240, 3030, 2925, 1652, 1617, 1268, 1079, 730;¹H NMR(DMSO-d₆), δ:9.62(s, 2H, N＝C—H), 7.87(d, J=8.83 Hz, 4H, PhH), 7.62~7.54(m, 4H, PhH), 7.30(d, J=8.85 Hz, 4H, PhH), 7.22~7.18(m, 4H, PhH), 5.19(s, 4H, CH₂), 2.52(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.58, 162.33, 160.45, 150.37, 129.73, 129.07, 106.34, 75.87, 56.44, 37.69, 28.86, 25.42, 21.13, 16.38;HRMS(positive-SIMS)计算值C₃₆H₃₀Cl₄N₁₀S₂[M+1]:806.0850，实测值:806.0859。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-甲氧基亚甲基氨基)-3-(4-氯苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM1b) 白色固体, 收率69.4%, mp 254.6~255.9 ℃, IR(KBr), σ/cm^-1:3340, 3124, 3030, 2925, 1652, 1617, 1268, 1145, 830;¹H NMR(DMSO-d₆), δ:9.62(s, 2H, N＝C—H), 7.87(d, J=8.83 Hz, 4H, PhH), 7.62~7.54(m, 4H, PhH), 7.30(d, J=8.85 Hz, 4H, PhH), 7.22~7.18(m, 4H, PhH), 5.19(s, 4H, CH₂), 3.38(s, 6H, CH₂), 2.52(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz，DMSO-d₆), δ:163.34, 162.68, 161.35, 151.87, 130.78, 125.43, 115.32, 68.79, 57.44, 52.36, 34.56, 26.18, 25.56, 22.00, 14.81;HRMS(positive-SIMS)计算值C₃₈H₃₆Cl₂N₁₀O₂S₂[M+1]:798.1841，实测值:798.1834。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-二茂铁基亚氨基-3-(4-氯苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM1c) 橘黄色固体, 收率60.7%, mp 287.3~288.5 ℃, IR(KBr), σ/cm^-1:3310, 3030, 2928, 1660, 1609, 1412, 1294, 1107, 783, 498;¹H NMR(DMSO-d₆), δ:9.52(s, 2H, N＝C—H), 7.67(d, J=8.4 Hz, 4H, PhH), 7.20(d, J=8.3 Hz, 4H, PhH), 5.17(s, 4H, CH₂), 4.79(t, J=1.5 Hz, 4H, FcH), 4.35(t, J=1.5 Hz, 4H, FcH), 4.28(s, 10H, FcH), 2.62(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.88, 143.14, 136.57, 133.28, 130.43, 128.41, 125.66, 122.30, 120.39, 120.08, 68.71, 51.41;HRMS(positive-SIMS)计算值C₄₈H₄₀N₁₀S₂Cl₂Fe₂[M+1]:955.1033, 实测值:955.1035。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-(吡啶-2-基亚氨基)-3-(4-氯苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM1d) 淡黄色固体, 收率67.5%, mp 214.3~215.6 ℃, IR(KBr), σ/cm^-1:3240, 3029, 2942, 1654, 1612, 1271, 1069, 722;¹H NMR(DMSO-d₆), δ:9.70(s, 2H, N＝C—H), 8.51(d, J=4.8 Hz, 2H, PyH), 8.19(d, J=7.9 Hz, 2H, PyH), 8.11(dd, J=1.3 Hz, J=7.7 Hz, 2H, PyH), 7.72(dd, J=1.4 Hz, J=7.01 Hz, 2H, PyH), 7.61~7.52(m, 8H, PhH), 5.39(s, 4H, CH₂), 2.54(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.65, 150.73, 143.62, 140.28, 135.41, 130.18, 126.09, 122.41, 120.54, 68.56, 50.58);HRMS(positive-SIMS)计算值C₃₈H₃₀Cl₂N₁₀O₂S₂[M+1]:740.1535, 实测值:740.1538。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-(呋喃-2-基亚氨基)-3-(4-氯苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM1e) 淡灰色固体, 收率67.9%, mp 231.6~233.2 ℃, IR(KBr), σ/cm^-1:3210, 3028, 2952, 1660, 1612, 1268, 1157, 1078, 764;¹H NMR(DMSO-d₆), δ:9.58(s, 2H, N＝C—H), 8.17(s, 2H, furan-H), 7.79(d, J=8.9 Hz, 4H, PhH), 7.42(d, J=3.6 Hz, 4H, PhH), 7.22(d, J=8.9 Hz, 2H, furan-H), 6.84(dd, J=1.71 Hz, J=3.6 Hz, 2H, furan-H), 5.17(s, 4H, CH₂), 2.70(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.65, 143.44, 140.12, 138.30, 136.58, 129.94, 125.75, 122.46, 120.54, 116.34, 110.63, 69.18, 51.22;HRMS(positive-SIMS)计算值C₃₆H₂₈Cl₂N₁₀O₂S₂[M+1]:718.1215, 实测值:718.1218。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-(噻吩-2-基亚氨基)-3-(4-氯苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM1f) 灰色固体, 收率70.1%, mp 226.4~227.5 ℃, IR(KBr), σ/cm^-1:3220, 3030, 2925, 1652, 1617, 1268, 1079, 730;¹H NMR(DMSO-d₆), δ:9.62(s, 2H, N＝C—H), 7.87(d, J=8.83 Hz, 4H, PhH), 7.30(d, J=8.85 Hz, 4H, PhH), 5.19(s, 4H, CH₂), 2.52(s, 8H, CH₂NCH₂), 7.64~7.63(m, 1H, thiophthene), 7.41(d, J=3.05 Hz, 1H, thiophthene), 7.04~7.03(m, 1H, thiophthene); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.11, 153.44, 149.62, 144.39, 136.88, 129.94, 128.75, 123.06, 119.44, 115.76, 113.41, 70.10, 61.62;HRMS(positive-SIMS)计算值C₃₂H₂₈Cl₂N₁₀S₄[M+1]:750.0758, 实测值:750.0749。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-氯苯基亚甲基氨基)-3-(4-甲氧基苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM2a) 白色固体, 收率62.5%, mp 246.4~248.1 ℃, IR(KBr), σ/cm^-1:3280, 3125, 3030, 2889, 1653, 1622, 1291, 1168, 845;¹H NMR(DMSO-d₆), δ:9.42(s, 2H, N＝C—H), 7.91(d, J=8.6 Hz, 4H, PhH), 7.62~7.57(m, 4H, PhH), 7.28(d, J=8.6 Hz, 4H, PhH), 7.19~7.15(m, 4H, PhH), 5.25(s, 4H, CH₂), 3.24(s, 6H, CH₂), 2.51(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz, CDCl₃), δ:162.47, 161.98, 160.43, 153.23, 130.73, 128.54, 108.39, 85.89, 56.67, 39.65, 31.34, 26.41, 23.12, 17.38;HRMS(positive-SIMS)计算值C₃₈H₃₆Cl₂N₁₀O₂S₂[M+1]:798.1841, 实测值:798.1832。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-甲氧基苯基亚甲基氨基)-3-(4-甲氧基苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM2b) 白色固体, 收率65.5%, mp 252.1~261.1 ℃, IR(KBr), σ/cm^-1:3340, 3120, 3025, 2880, 1642, 1629, 1281, 1169, 850;¹H NMR(DMSO-d₆), δ:9.42(s, 2H, N＝C—H), 7.91(d, J=8.6 Hz, 4H, PhH), 7.62~7.57(m, 4H, PhH), 7.28(d, J=8.6 Hz, 4H, PhH), 7.19~7.15(m, 4H, PhH), 5.25(s, 4H, CH₂), 3.24(s, 6H, CH₂), 3.22(s, 6H, CH₂) 2.51(s, 8H, CH₂NCH₂); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.56, 144.13, 136.24, 133.67, 130.43, 127.49, 125.14, 122.47, 121.39, 120.78, 69.74, 52.01;HRMS(positive-SIMS)计算值C₄₀H₄₂N₁₀O₄S₂[M+1]:790.2832, 实测值:790.2830。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-二茂铁基亚氨基)-3-(4-甲氧基苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM2c) 褐色固体, 收率为62.7%, mp 210.2~211.5 ℃, IR(KBr), σ/cm^-1:3310, 3125, 3029, 1650, 1410, 1264, 1125, 824, 498;¹H NMR(DMSO-d₆), δ:9.52(s, 2H, N＝CH), 7.54(d, J=8.3 Hz, 4H, PhH), 7.13(d, J=8.3 Hz, 4H, PhH), 5.14(s, 4H, CH₂), 4.81 (t, J=1.6 Hz, 4H, CH₂), 4.30 (t, J=1.6 Hz, 4H, CH₂), 4.24(d, 10H, FcH), 3.81(s, 6H, FcH), 2.62(s, 8H, CH₂); ¹³C NMR(125 MHz, DMSO-d₆), δ:162.57, 143.53, 136.14, 134.67, 131.43, 129.03, 125.56, 123.00, 121.33, 120.58, 69.61, 51.81;HRMS(positive-SIMS)计算值C₄₄H₄₅Fe₂N₁₀O₂S₂[M+1]:932.1789, 实测值:932.1793。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-(吡啶-2-基亚氨基)-3-(4-甲氧基苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM2d) 黄色固体, 收率60.4%, mp 217.3~218.9 ℃, IR(KBr), σ/cm^-1:3210, 3030, 2920, 1620, 1510, 1251, 1153, 845, 822;¹H NMR(CDCl₃), δ:9.52(s, 2H, N＝C—H), 8.37(d, J=4.8 Hz, 2H, PhH), 8.19(d, J=7.9 Hz, 2H, PhH), 8.05(dd, J=1.28 Hz, J=7.8 Hz, 2H, PhH), 7.73(dd, J=1.44 Hz, J=6.9 Hz, 4H, PhH), 7.58~7.34(m, 6H, PhH), 5.42(s, 4H, CH₂), 3.57(s, 6H, CH₂); ¹³C NMR(125 MHz, CDCl₃), δ:162.45, 150.67, 143.35, 140.98, 135.64, 131.16, 127.84, 122.23, 121.84, 68.46, 50.79;HRMS(positive-SIMS)计算值C₃₆H₃₆N₁₂O₂S₂[M+1]:732.2526, 实测值:732.2518。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-(呋喃-2-基亚氨基)-3-(4-甲氧基苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM2e) 灰色固体, 收率59.8%, mp 201.7~203.2 ℃, IR(KBr), σ/cm^-1:3210, 3030, 2951, 1659, 1609, 1277, 1154, 852, 769;¹H NMR(CDCl₃), δ:9.58(s, 2H, N＝C—H), 8.10(s, 2H, PhH), 7.85(d, J=8.9 Hz, 4H, PhH), 7.38(d, J=3.5 Hz, 2H, PhH), 7.22(d, J=8.7 Hz, 4H, PhH), 6.79(dd, J=1.6 Hz, J=3.4 Hz, 2H, PhH), 5.17(s, 4H, CH₂), 3.79(s, 6H, CH₂), 2.70(s, 8H, CH₂); ¹³C NMR(125 MHz, CDCl₃), δ:162.78, 144.67, 141.02, 139.00, 136.98, 130.04, 125.35, 122.38, 121.74, 116.47, 111.73, 69.59, 52.01;HRMS(positive-SIMS)计算值C₃₄H₃₄N₁₀O₄S₂[M+1]:710.2206, 实测值:710.2218。

1, 1′-(哌嗪-1, 4-双(亚甲基))-4-(噻吩-2-基亚氨基)-3-(4-甲氧基苯基)-1H-1, 2, 4-三唑-5(4H)-硫酮(TM2f) 浅灰色固体, 收率58.9%, mp 203.8~204.6 ℃, IR(KBr), σ/cm^-1:3218, 3030, 2943, 1660, 1607, 1276, 1158, 858, 764;¹H NMR(CDCl₃), δ:9.58(s, 2H, N＝C—H), 8.10(s, 2H, PhH), 7.85(d, J=8.9 Hz, 4H, PhH), 7.38(d, J=3.5 Hz, 2H, PhH), 7.22(d, J=8.7 Hz, 4H, PhH), 6.79(dd, J=1.6 Hz, J=3.4 Hz, 2H, thiophthene), 5.17(s, 4H, CH₂), 3.79(s, 6H), 2.70(s, 8H); ¹³C NMR(125 MHz, CDCl₃), δ:162.71, 152.84, 149.82, 144.47, 136.28, 129.85, 129.05, 123.06, 118.44, 116.76, 114.81, 70.35, 60.62;HRMS(positive-SIMS)计算值C₃₄H₃₄N₁₀O₂S₄[M+1]:742.1749, 实测值:742.1742。

1, 1′-(哌嗪-1, 4-双(亚甲基))双(4-(4-氯苯烯基氨基)-3-戊基-1H-1, 2, 4-三唑-5(4H)-硫酮)(TM3a) 白色粉末, 收率90.2%, mp 204.1~205.2 ℃, IR(KBr), σ/cm^-1:3260, 2931, 2856, 1587, 1435, 1399, 1319, 1275, 1175, 1087, 1005, 969, 825, 737;¹H NMR(CDCl₃), δ:10.51(s, 2H, N＝C—H), 7.80(d, J=8.5 Hz, 4H, PhH), 7.00(d, J=8.5 Hz, 4H, furan-H), 5.11(s, 4H, CH₂), 2.85(s, 8H, CH₂NCH₂), 2.82~2.76(m, 4H, CH₂), 1.76~1.72(m, 4H, CH₂), 1.40~1.26(m, 8H, CH₂), 0.92~0.89(m, 6H, CH₃); ¹³C NMR(125 MHz, CDCl3), δ:162.56, 162.53, 161.55, 150.87, 130.73, 129.07, 116.34, 68.87, 50.44, 31.28, 25.86, 25.06, 22.13, 13.83;HRMS(positive-SIMS)计算值C₃₄H₄₄Cl₂N₁₀S₂[M+1]:735.3365, 实测值:725.3369。

1, 1′-(哌嗪-1, 4-双(亚甲基))双(4-(4-甲氧苯烯基氨基)-3-戊基-1H-1, 2, 4-三唑-5(4H)-硫酮)(TM3b) 白色粉末, 收率88.6%, mp 184.1~185.2 ℃, IR(KBr), σ/cm^-1:3240, 2931, 2803, 1605, 1507, 1409, 1355, 1283, 1167, 1015, 835;¹H NMR(CDCl₃), δ:10.13(s, 2H, N＝C—H), 7.82(d, J=8.8 Hz, 4H, PhH), 7.00(d, J=8.8 Hz, 4H, PhH), 5.10(s, 4H, CH₂), 3.88(s, 6H, OCH₃), 2.86(s, 8H, CH₂NCH₂), 2.78(t, J=7.6 Hz, 4H, CH₂), 1.76~1.72(m, 4H, CH₂), 1.38~1.35(m, 8H, CH₂), 0.91~0.89(m, 6H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:163.06, 162.61, 161.46, 150.84, 130.53, 125.32, 114.42, 68.77, 55.46, 50.46, 31.24, 25.78, 25.00, 22.10, 13.80;HRMS(positive-SIMS)计算值C₃₆H₅₀N₁₀O₂S₂[M+1]:727.4356, 实测值:727.4351。

1, 1′-(哌嗪-1, 4-双(亚甲基))双(4-二茂铁基-3-十一烷基-1H-1, 2, 4-三唑-5(4H)-硫酮)(TM3c) 橘黄色粉末, 收率75.6%, mp 186.8~188.2 ℃, IR(KBr), σ/cm^-1:3210, 2936, 2812, 2678, 1605, 1417, 1355, 1286, 1167, 1015, 835;¹H NMR(CDCl₃), δ:9.71(s, 1H, N＝CH), 5.11(s, 2H, CH₂), 4.76(t, J=1.8 Hz, 2H, FcH), 4.57(t, J=1.8 Hz, 2H, FcH), 4.35(s, 5H, FcH), 2.86(s, 4H, CH₂NCH₂), 2.79(t, J=7.6 Hz, 2H, CH₂), 1.76~1.72(m, 2H, CH₂), 1.39~1.36(m, 4H, CH₂), 0.92~0.89(m, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:166.53, 162.55, 150.52, 75.57, 71.95, 70.42, 69.01, 68.86, 50.46, 31.23, 25.81, 25.00, 22.16, 13.87;HRMS(positive-SIMS)计算值C₄₆H₆₆Fe₂N₁₀S₂[M+1]:934.3612, 实测值:934.3621。

1, 1′-(哌嗪-1, 4-双(亚甲基))二(3-戊基-4-(吡啶-2-基甲烯基氨基)-1H-1, 2, 4-三唑-5(4H)-硫酮)(TM3d) 淡黄色粉末, 收率67.9%, mp 188.9~190.2 ℃, IR(KBr), σ/cm^-1:3310, 2952, 2844, 1675, 1607, 1484, 1346, 1267, 1170, 1060, 855;¹H NMR(CDCl₃), δ:10.67(s, 1H, N＝CH), 8.09(d, J=8.0 Hz, 1H, PyH), 7.84~7.80(m, 1H, PyH), 7.75~7.74(m, 1H, PyH), 7.41~7.39(m, 1H, PyH), 5.10(s, 2H, CH₂), 2.88(s, 4H, CH₂NCH₂), 2.80 (t, J=7.6 Hz, 2H, CH₂), 1.75~1.70(m, 2H, CH₂), 1.38~1.35(m, 2H, CH₂), 1.31~1.25(m, 14H, CH₂), 0.89 (t, J=6.9 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:162.52, 161.51, 160.05, 150.87, 150.15, 136.54, 125.68, 121.65, 68.83, 50.45, 31.22, 25.87, 25.00, 22.15, 13.87;HRMS(positive-SIMS)计算值C₃₂H₄₄N₁₂S₂[M+1]:697.4363, 实测值:697.4355。

1, 1′-(哌嗪-1, 4-双(亚甲基))双(4-(呋喃-2-基甲烯基氨基)-3-十一烷基-1H-1, 2, 4-三唑-5(4H)-硫酮)(TM3e) 白色粉末, 收率78.2%, mp 169.8~170.8 ℃, IR(KBr), σ/cm^-1:3320, 2921, 2864, 1667, 1599, 1505, 1443, 1381, 1319, 1275, 1229, 1157, 1069, 1005, 943, 845, 719;¹H NMR(CDCl₃), δ:10.30(s, H, N＝C—H), 7.66(d, J=1.4 Hz, 1H, furan-H), 7.04(d, J=3.5 Hz, 1H, furan-H), 6.60(dd, J=1.8, 3.5 Hz, 1H, furan-H), 5.13(s, 2H, CH₂), 2.86(s, 4H, CH₂NCH₂), 1.77~1.73(m, 2H, CH₂), 1.39~1.35(m, 2H, CH₂), 1.31~1.26(m, 14H, CH₂), 0.92 (t, J=7.0 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:162.57, 161.50, 150.84, 149.67, 146.67, 118.29, 112.48, 68.84, 50.46, 31.26, 25.87, 25.08, 22.15, 13.85;HRMS(positive-SIMS)计算值C₃₀H₄₂N₁₀O₂S₂[M+1]:638.2934, 实测值:638.2939。

1, 1′-(哌嗪-1, 4-双(亚甲基))双(4-(噻吩-2-基甲烯基氨基)-3-十一烷基-1H-1, 2, 4-三唑-5(4H)-硫酮)(TM3f) 白色粉末, 收率79.3%, mp 166.9~167.8 ℃, IR(KBr), σ/cm^-1:3020, 2920, 2872, 1668, 1587, 1506, 1442, 1388, 1313, 1269, 1238, 1156, 1068, 1015, 940, 848, 718;¹H NMR(CDCl₃), δ:10.30(s, H, N＝C—H), 7.66(d, J=1.4 Hz, 1H, furan-H), 7.04(d, J=3.5 Hz, 1H, furan-H), 6.60(dd, J=1.8, 3.5 Hz, 1H, furan-H), 5.13(s, 2H, CH₂), 2.86(s, 4H, CH₂NCH₂), 1.77~1.73(m, 2H, CH₂), 1.39~1.35(m, 2H, CH₂), 1.31~1.26(m, 14H, CH₂), 0.92 (t, J=7.0 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:162.56, 152.87, 149.47, 144.54, 136.68, 129.45, 129.48, 124.66, 116.47, 115.73, 112.91, 70.85, 62.02;HRMS(positive-SIMS)计算值C₃₀H₄₂N₁₀S₄[M+1]:670.2477, 实测值:670.2470。

5, 5′-((6-吗啉-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(4-甲氧苯亚基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM4a) 黄色固体, 收率:69.6%, mp 198.3~199.9 ℃, IR(KBr), σ/cm^-1:3260, 3050, 2930, 2850, 1685, 1600, 1500, 1450, 1380, 1290, 1250, 750;¹H NMR(CDCl₃), δ:8.55(s, 1H, N—CH), 7.76(d, J=8.80 Hz, 2H, PhH), 7.70(t, J=8.80 Hz, 2H, PhH), 3.89(s, 3H, OCH₃), 3.73(t, J=5.00 Hz, 2H, CH₂OCH₂), 3.55(t, J=4.75 Hz, 2H, CH₂NCH₂), 2.85(t, J=7.20 Hz, 2H, CH₂), 1.81(t, J=7.20 Hz, 2H, CH₂), 1.36(t, J=7.00 Hz, 4H, CH₂), 0.89(t, J=7.00 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.64, 163.04, 162.60, 161.46, 160.78, 150.80, 130.52, 125.30, 114.00, 65.14, 55.45, 43.20, 31.20, 25.74, 25.00, 22.14, 13.84;HRMS(positive-SIMS)计算值C₃₇H₄₆N₁₂O₃S₂[M+1]:770.3257, 实测值:770.3250。

5, 5′-((6-吗啉-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-二茂铁基)3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM4b) 橘红色固体, 收率75.8%, mp 205.9~207.1 ℃, IR(KBr), σ/cm^-1:3270, 3080, 2935, 2820, 1680, 1292, 1108, 840;¹H NMR(CDCl₃), δ:8.23(s, 1H, CH=N), 4.89(s, 2H, FcH), 4.59(s, 2H, FcH), 4.25(s, 5H, FcH), 3.73(t, J=5.00 Hz, 2H, CH₂OCH₂), 3.55(t, J=4.75 Hz, 2H, CH₂NCH₂), 2.85(t, J=7.20 Hz, 2H, CH₂), 1.81(t, J=7.20 Hz, 2H, CH₂), 1.36(t, J=7.00 Hz, 4H, CH₂), 0.90(t, J=7.00 Hz, 3H, CH₃); ³C NMR(125 MHz, CDCl₃), δ:176.70, 166.52, 162.50, 160.70, 150.80, 75.50, 71.96, 70.40, 69.06, 66.90, 65.18, 43.20, 31.20, 25.80, 25.01, 22.14, 13.85;HRMS(positive-SIMS)计算值C₄₃H₅₀Fe₂N₁₂OS₂[M+1]:926.2371, 实测值:926.2365。

5, 5′-((6-吗啉-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-吡啶基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM4c) 黄绿色固体, 收率70.3%, mp 196.5~197.9 ℃, IR(KBr), σ/cm^-1:3240, 3050, 2940, 2830, 1690, 1600, 1500, 1380, 1286, 1240, 745;¹H NMR(CDCl₃), δ:8.76~8.74(m, 1H, PyH), 8.30(s, 1H, N—CH), 8.09(d, J=7.95 Hz, 1H, PyH), 7.83~7.80(m, 1H, PyH), 7.42-7.40(m, 1H, PyH), 3.73(t, J=5.00 Hz, 2H, CH₂OCH₂), 3.55(t, J=4.75 Hz, 2H, CH₂NCH₂), 2.85(t, J=7.20 Hz, 2H, CH₂), 1.81(t, J=7.20 Hz, 2H, CH₂), 1.36(t, J=7.00 Hz, 4H, CH₂), 0.90(t, J=7.00 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.60, 163.14, 160.74, 160.00, 152.30, 151.00, 150.80, 136.54, 125.68, 121.70, 65.10, 43.20, 31.20, 25.80, 25.00, 22.14, 13.80;HRMS(positive-SIMS)计算值C₃₃H₄₀N₁₄OS₂[M+1]:712.2951, 实测值:712.2947。

5, 5′-((6-吗啉-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-呋喃基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM4d) 深棕色固体, 收率75.9%, mp 172.8~173.4 ℃, IR(KBr), σ/cm^-1:3240, 3160, 2950, 2860, 1690, 1380, 1270, 1240, 732;¹H NMR(CDCl₃), δ:8.31(s, 1H, CH=N), 7.67(d, J=6.95 Hz, 1H, furan-H), 7.06(t, J=6.95 Hz, 1H, furan-H), 6.59(q, J=6.95 Hz, 1H, 呋喃环), 3.74(t, J=5.00 Hz, 2H, CH₂OCH₂), 3.55(t, J=4.75 Hz, 2H, CH₂NCH₂), 2.85(t, J=7.20 Hz, 2H, CH₂), 1.81(t, J=7.20 Hz, 2H, CH₂), 1.36(t, J=7.00 Hz, 4H, CH₂), 0.89(t, J=7.00 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.60, 162.50, 160.74, 160.00, 150.88, 149.26, 148.16, 146.65, 118.30, 112.52, 65.10, 43.20, 31.25, 28.94, 25.02, 22.14, 13.80;HRMS(positive-SIMS)计算值C₃₁H₃₈N₁₂O₃S₂[M+1]:690.2631, 实测值:690.2628。

5, 5′-((6-吗啉-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-噻吩基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM4e) 褐色固体, 收率77.9%, mp 175.7~176.3 ℃, IR(KBr), σ/cm^-1:3260, 3170, 2960, 2880, 1790, 1390, 1370, 1240, 737;¹H NMR(CDCl₃), δ:8.31(s, 1H, CH＝N), 7.64~7.63(m, 1H, 噻吩), 7.41(d, J=3.05 Hz, 1H, 噻吩), 7.04~7.03(m, 1H, 噻吩), 3.74(t, J=5.00 Hz, 2H, CH₂OCH₂), 3.55(t, J=4.75 Hz, 2H, CH₂NCH₂), 2.78 (t, J=7.6 Hz, 2H, CH₂), 1.76~1.70(m, 2H, CH₂), 1.40~1.35(m, 2H, CH₂), 1.31~1.25(m, 14H, CH₂), 0.89 (t, J=6.8 Hz, 3H, CH₃); ³C NMR(125 MHz, CDCl₃), δ:166.20, 152.59, 160.74, 150.50, 144.88, 13916, 138.05, 136.61, 132.30, 129.72, 108.80, 83.20, 61.95, 48.94, 35.06, 22.32, 19.60;HRMS(positive-SIMS)计算值C₄₃H₆₂N₁₂OS₄[M+1]:890.4052, 实测值:890.4057。

5, 5′-((6-吡咯烷-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(4-对甲氧苯亚基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM5a) 淡黄色固体, 收率77.3%, mp 211.1~212.8 ℃, IR(KBr), σ/cm^-1:3220, 3030, 2960, 2854, 2820, 1690, 1606, 1520, 1450, 1380, 1290, 1245, 830, 730;¹H NMR(CDCl₃), δ:8.24(s, 1H, N—CH), 7.76(d, J=8.80 Hz, 2H, PhH), 7.70(t, J=8.80 Hz, 2H, PhH), 3.89(s, 1H, OCH₃), 3.73(t, J=4.95 Hz, 2H, CH₂N), 3.55(t, J=4.95 Hz, 2H, CH₂), 2.85(t, J=7.50 Hz, 2H, CH₂), 1.81(t, J=7.45 Hz, 2H, CH₂), 1.38~1.32(m, 4H, CH₂), 0.90(q, J=7.50 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.64, 163.04, 162.60, 161.46, 160.78, 150.80, 130.52, 125.30, 114.00, 54.17, 55.45, 31.20, 25.74, 25.24, 25.00, 22.14, 13.84;HRMS(positive-SIMS)计算值C₃₇H₄₆N₁₂O₂S₂[M+1]:754.3308, 实测值:745.3314。

5, 5′-((6-吡咯烷-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-二茂铁基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM5b) 红色固体, 收率72.6%, mp 165.3~166.7 ℃, IR(KBr), σ/cm^-1:3250, 3080, 2930, 2820, 1680, 1380, 1292, 1106, 835;¹H NMR(CDCl₃), δ:8.24(s, 1H, CH＝N), 4.90(t, J=3.60 Hz, 2H, FcH), 4.60(t, J=3.60 Hz, 2H, FcH), 4.26(d, J=3.60 Hz, 5H, FcH), 3.73(t, J=4.95 Hz, 2H, CH₂N), 3.55(t, J=5.90 Hz, 2H, CH₂), 2.85(t, J=7.50 Hz, 2H, CH₂), 1.81(t, J=7.45 Hz, 2H, CH₂), 1.38~1.32(m, 4H, CH₂), 0.90(q, J=7.50 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.70, 166.52, 162.50, 160.70, 150.80, 77.01, 76.75, 75.50, 71.96, 70.40, 69.06, 65.00, 54.16, 31.20, 25.80, 25.24, 25.01, 22.14, 13.85;HRMS(positive-SIMS)计算值C₄₃H₅₀Fe₂N₁₂S₂[M+1]:910.2422, 实测值:910.2420。

5, 5′-((6-吡咯烷-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-吡啶基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM5c) 黄绿色固体, 收率71.6%, mp 254.1~-255.9 ℃, IR(KBr), σ/cm^-1:3240, 3050, 2940, 2830, 1690, 1600, 1380, 1270, 885, 745;¹H NMR(CDCl₃), δ:8.24(s, 1H, N—CH), 8.09(d, J=7.95 Hz, 1H, PyH), 7.83~7.80(m, 1H, PyH), 7.42~7.40(m, 1H, PyH), 3.73(t, J=4.95 Hz, 2H, CH₂N), 3.55(t, J=4.95 Hz, 2H, CH₂), 2.85(t, J=7.50 Hz, 2H, CH₂), 1.81(t, J=7.45 Hz, 2H, CH₂), 1.38~1.32(m, 4H, CH₂), 0.89(q, J=7.50 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.60, 163.14, 160.74, 160.00, 152.30, 151.00, 150.80, 136.54, 125.68, 121.70, 54.15, 31.20, 25.80, 25.26, 25.00, 22.14, 13.80;HRMS(positive-SIMS)计算值C₃₃H₄₀N₁₄S₂[M+1]:696.3002, 实测值:696.3004。

5, 5′-((6-吡咯烷-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-呋喃基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM5d) 深棕色固体, 收率65.8%, mp 178.4~179.6 ℃, IR(KBr), σ/cm^-1:3210, 2950, 2860, 1690, 1600, 1380, 1270, 965, 732;¹H NMR(CDCl₃), δ:8.24(s, 1H, -CH=N-), 7.67(d, J= 6.95 Hz, 1H, furan-H), 7.06(t, J=6.95 Hz, 1H, furan-H), 6.59(q, J=6.95 Hz, 1H, furan-H)3.73(t, J=4.95 Hz, 2H, CH₂N), 3.55(t, J=4.95 Hz, 2H, CH₂), 2.85(t, J=7.50 Hz, 2H, CH₂), 1.81(t, J=7.45 Hz, 2H, CH₂), 1.38~1.32(m, 4H, CH₂), 0.89(q, J=7.50 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.60, 162.50, 160.74, 160.00, 150.88, 149.26, 148.16, 146.65, 118.30, 112.52, 54.14, 31.25, 28.94, 25.26, 25.02, 22.14, 13.80;HRMS(positive-SIMS)计算值C₃₁H₃₈N₁₂O₂S₂[M+1]:674.2682, 实测值:674.2690。

5, 5′-((6-吡咯烷-1, 3, 5-三嗪-2, 4-二基)双(硫醚))双(N-(2-噻吩基)-3-正戊基-4H-1, 2, 4-三唑-4-胺)(TM5e) 棕色固体, 收率68.8%, mp 168.2~169.4 ℃, IR(KBr), σ/cm^-1:3230, 3110, 2950, 2860, 1690, 1600, 1380, 1270, 965, 732;¹H NMR(CDCl₃), δ:8.24(s, 1H, N—CH), 7.64~7.63(m, 1H, thiophthene), 7.41(d, J=3.05 Hz, 1H, thiophthene), 7.04~7.03(m, 1H, thiophthene) 3.73(t, J=4.95 Hz, 2H, CH₂N), 3.55(t, J=4.95 Hz, 2H, CH₂), 2.78 (t, J=7.6 Hz, 2H, CH₂), 1.76~1.70(m, 2H, CH₂), 1.40~1.35(m, 2H, CH₂), 1.31~1.25(m, 14H, CH₂), 0.89 (t, J=6.8 Hz, 3H, CH₃); ¹³C NMR(125 MHz, CDCl₃), δ:176.57, 163.13, 162.14, 160.64, 149.87, 149.26, 144.16, 137.65, 135.30, 112.34, 54.19, 31.65, 28.75, 26.96, 25.67, 22.14, 15.32;HRMS(positive-SIMS)计算值C₄₃H₆₂N₁₂S₄[M+1]:874.4103, 实测值:874.4111。

2 结果与讨论

2.1 目标分子的表征

在IR谱图中，目标分子在3210~3420 cm^-1宽峰为—NH—吸收峰，在3030 cm^-1左右为苯环上C—H的吸收峰，1650和1500 cm^-1左右出现C＝N和C＝C的吸收峰，在1290和1150 cm^-1左右分别为C—N和C—O的吸收峰；¹H NMR谱图中，—NH—上的质子信号在9.62左右处，在7.96和6.87处为苯环的质子信号，在3.70和3.60左右处出现的为吗啉的质子信号，在2.50和1.95~1.91左右出现吡咯的质子信号；在HRMS谱图中，所有化合物均出现[M+1]⁺峰，表明成功合成了目标分子。

2.2 目标分子对Cdc25B的抑制活性测试

目标分子初筛选择质量浓度均为5 mg/L，Cdc25B抑制活性的阳性对照物为Na₃VO₄。选择抑制率大于50%的化合物进行复筛，得出抑制活性剂量依赖关系，即半抑制浓度IC₅₀(Half maximal inhibitory concentration)值。抑制活性筛选实验参照文献[25]的方法进行。本实验由国家新药中心协助完成，测试结果见表 1所示。

表 1 目标分子(TM)对Cdc25B的抑制活性 Table 1. Inhibitory activities of target molecules against Cdc25B

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Compd.	Inhibitory rate/%	IC₅₀^a/(mg·L^-1)
TM1a	58.78	3.24±0.17
TM1b	21.59	NA
TM1c	32.96	NA
TM1d	41.59	NA
TM1e	97.75	0.66±0.08
TM1f	48.67	NA
TM2a	57.37	3.11±0.25
TM2b	56.31	2.93±0.32
TM2c	88.21±0.13	1.17±0.11
TM2d	75.73±0.21	2.35±0.41
TM2e	43.84±1.04	NA
TM2f	69.35±1.02	2.06±0.07
TM3a	40.59±0.37	NA
TM3b	37.59±0.17	NA
TM3c	89.47±1.26	1.49±0.15
TM3d	94.54±0.07	1.53±0.06
TM3e	60.57±0.09	1.96±0.08
TM3f	90.13±0.37	2.04±0.72
TM4a	64.16±1.01	2.69±0.13
TM4b	97.91±0.15	5.71±1.21
TM4c	86.71±3.90	2.75±0.22
TM4d	93.03±0.60	1.04±0.26
TM4e	91.85±0.14	1.73±0.23
TM5a	20.38±3.45	NA
TM5b	98.14±0.04	1.11±0.56
TM5c	99.45±0.07	3.75 ± 0.84
TM5d	68.50±6.91	1.17 ± 0.15
TM5e	87.64±1.12	1.74±0.41
C	70.53±0.52	1.06±0.09
D	70.41±2.38	1.31±0.12
Positive control:Na₃VO₄ was for Cdc25B(IC₅₀=(1.86±0.244) mg/L)
a.Value tested at 5 mg/L mass concentration.

2.3 目标分子的构效分析

总体比较两类目标分子对Cdc25B的抑制活性测试结果发现，绝大多数以均三嗪为桥对接的目标分子表现出更好的活性，原因可能是该类分子与生物大分子相似度大，更易发生相互作用而使活性得到提高。分析以均三嗪为桥对接的分子，首先，当4-氨基未被修饰时(C和D)，抑制活性较好，原因可能是氨基与生物大分子之间产生了氢键，其次，利用四氢吡咯比吗啉修饰时结果更好。分析以哌嗪为桥对接的分子，首先3-正戊基比3-对氯苯基和3-对甲氧苯基修饰的目标分子抑制活性优良，原因可能是目标分子的脂溶性得到提高；其次对比对氯苯基和对甲氧苯基分别对3-号位和4-号位氨基修饰的结果发现，当两个位点采用同种基团修饰时(TM1a, TM2b)，比二者交替修饰时(TM1b, TM2a)效果更好，这可能是因为前者对1, 2, 4-三唑环上电子偏移作用相同，电子分布合理；利用4-号位氨基构筑希夫碱时发现，引入呋喃、噻吩和吡啶等杂环时，均取得优良的抑制效果，原因可能是其中的杂原子与生物大分子产生了氢键作用。由此可见，通过对1, 2, 4-三唑不同位点进行结构修饰，发挥了活性叠加的作用，可筛选出对Cdc25B具有优良抑制活性的分子，目标分子可作为为抗癌药物筛选的重要研究对象。

3 结论

根据许多天然产物和临床药物分子富含杂环活性组块的结构特点，本文首次通过简捷、经典的合成策略，将1, 2, 4-三唑、哌嗪和均三唑等药效基团成功构筑与同一分子中, 为研究构效关系，分别对1, 2, 4-三唑骨架中的三个位点进行了修饰，发现绝大多数目标分子显示了优良的Cdc25B抑制活性。

所设计的目标分子均具有近Ⅴ型结构特征，分子结构具有一定柔韧性，这一新型骨架结构为目标分子与生物大分子更好的相互作用创造了有利的条件，尤其是双1, 2, 4-三唑组快的构建，形成了两种对称的“钳式”结构，这种双方位、多杂原子共存的结构特征，使目标分子更易通过氢键作用“抓住”生物大分子，从而产生优异的药理活性，为新型Cdc25B抑制剂的设计与合成提供了重要的参考。本文设计合成的许多目标分子有望作为Cdc25B抑制剂，成为抗肿瘤药物先导物，所得结果达到了预期研究目的。

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Scheme 1 Synthetic routes of target molecules

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表 1 目标分子(TM)对Cdc25B的抑制活性

Table 1. Inhibitory activities of target molecules against Cdc25B

Compd.	Inhibitory rate/%	IC₅₀^a/(mg·L^-1)
TM1a	58.78	3.24±0.17
TM1b	21.59	NA
TM1c	32.96	NA
TM1d	41.59	NA
TM1e	97.75	0.66±0.08
TM1f	48.67	NA
TM2a	57.37	3.11±0.25
TM2b	56.31	2.93±0.32
TM2c	88.21±0.13	1.17±0.11
TM2d	75.73±0.21	2.35±0.41
TM2e	43.84±1.04	NA
TM2f	69.35±1.02	2.06±0.07
TM3a	40.59±0.37	NA
TM3b	37.59±0.17	NA
TM3c	89.47±1.26	1.49±0.15
TM3d	94.54±0.07	1.53±0.06
TM3e	60.57±0.09	1.96±0.08
TM3f	90.13±0.37	2.04±0.72
TM4a	64.16±1.01	2.69±0.13
TM4b	97.91±0.15	5.71±1.21
TM4c	86.71±3.90	2.75±0.22
TM4d	93.03±0.60	1.04±0.26
TM4e	91.85±0.14	1.73±0.23
TM5a	20.38±3.45	NA
TM5b	98.14±0.04	1.11±0.56
TM5c	99.45±0.07	3.75 ± 0.84
TM5d	68.50±6.91	1.17 ± 0.15
TM5e	87.64±1.12	1.74±0.41
C	70.53±0.52	1.06±0.09
D	70.41±2.38	1.31±0.12
Positive control:Na₃VO₄ was for Cdc25B(IC₅₀=(1.86±0.244) mg/L)
a.Value tested at 5 mg/L mass concentration.

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