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  有机农药作为外源化学物质进入环境之中, 不可避免地在环境的各组成要素之中迁移并被转化, 导致产生大量代谢产物, 相对于母体, 代谢产物往往会降低或丧失母体的活性, 但是一些代谢产物的活性比母体进一步提高:马拉硫磷的代谢物马拉氧磷对乙酰胆碱酯酶的抑制活性提高约2个数量级^[3], 噁唑禾草灵的代谢产物噁唑禾草灵酸对脂肪酸重新合成的抑制率比母体高10倍^[4], 噻虫嗪代谢产物噻虫胺比母体具有更好的杀虫活性^{[5, 6]}(图 1), 可见, 农药的代谢产物化学结构也可为新农药创制先导化合物的发现提供重要的启示.

  

  图2 设计目标化合物

  Figure 2. Target compound design

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  酰胺类化合物是医药、农药和兽药等研究领域中广受关注的化学结构, 往往具有独特的杀虫、杀菌、除草以及植物生长调节等活性^[7~15], 如苯霜灵、甲霜灵、呋霜灵等均是成功的商品化酰胺类杀菌剂品种(图 2), 其化学结构中都含有N-(2, 6-二甲基苯基)丙酸甲酯基团, 可见该结构对杀菌活性的发挥具有十分重要的作用.苯霜灵是一种高效、低毒、持效期长的内吸性杀菌剂, 能够抑制核糖体RNA的合成, 导致蛋白质合成减少, 从而抑制菌丝体的生长, 被广泛应用于防治霜霉病、早疫病和晚疫病等卵菌纲病害^{[16, 17]}.苯霜灵的代谢物主要有三个, 代谢物A、代谢物B和代谢物C^{[18, 19]}(图 2), 其中代谢物A保留了母体中的N-(2, 6-二甲基苯基)丙酸甲酯基药效团, 在国家自然基金重点项目“农药代谢物在水生环境及水生生物中的生成机制及健康效应研究”研究中, 课题组前期合成了苯霜灵的三个典型代谢物的标样, 生物活性筛选表明, 代谢物A其具有较好的杀菌活性, 其对辣椒疫霉和番茄晚疫病原菌的抑制率分别为50.3%和70.2%, 因此本文以苯霜灵代谢物A为先导, 通过结构优化设计合成系列未见文献报道的双酰胺类化合物(图 2), 其结构经过核磁氢谱、碳谱以及高分辨质谱的确认, 初步生物活性测试结果表明, 在50 mg/L浓度下, 目标化合物7d, 7e和7k有较好的杀菌活性, 目标化合物合成路线如Scheme 1所示.

  

  图图式 1 目标化合物合成路线

  Figure 图式 1. Synthetic route of target compound

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  图1 农药及其代谢产物

  Figure 1. Pesticides and their metabolites

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  20世纪30年代, 德国科学家偶然发现百浪多息在试管内无杀菌活性, 在生物体内却可以杀死细菌这一反常现象, 后研究证明百浪多息中的有效成分是其代谢产物对氨苯磺胺, 进而发现了磺胺类药物^[1]. 1940年, Wood和Fileds建立了代谢拮抗学说, 新颖的代谢物结构成为医药先导化合物的发现方法之一, 被广泛应用于医药行业, 如由阿司咪唑进一步发现诺阿司咪唑, 提高了对靶标受体的选择性, 增加了安全性^[2].

  1    结果与讨论

  1.1    合成讨论及波谱分析

  目标化合物的合成方法主要有两种: (1) 丙二酰氯选择性地分别和化合物1、中间体6反应; (2) 通过Scheme 3所示方法合成目标化合物.第一种方法反应过程不易跟踪监测, 易产生丙二酰氯与化合物1或中间体6的双取代产物, 难以与目标化合物7分离, 最后得到的产物成分复杂, 纯度差, 因此我们采用第二种方法合成目标化合物.

  中间体3合成过程中, 选择由氢氧化钠水解中间体2, 发现酯键和酰胺键均断裂, 产物很杂.选择由碱性较弱的氢氧化锂水解, 可以选择性地脱去酯基, 而不影响酰胺键, 得到高纯度中间体3.

  中间体2的合成方法主要有两种: (1) 丙二酸单甲酯, DCC缩合; (2) 丙二酸单甲酯酰氯, 三乙胺作缚酸剂进行酰胺化.第一种方法反应时间长, DCC较难除净, 给后处理带来很大不便, 得到的中间体2纯度较低.第二种方法在冰浴条件下, 将丙二酸单甲酯酰氯滴加到化合物1的二氯甲烷溶液中, 0.5 h即可反应完毕, 重结晶便可得到高纯度的中间体2, 因此选择第二种方法合成中间体2.

  合成目标化合物7时, 尝试了不同反应条件对收率的影响.当选用三乙胺作缚酸剂、二氯甲烷作溶剂时, 和吡啶作溶剂和缚酸剂时, 反应5 h并尝试了加热回流反应, 仍只有很少产物产生.选用甲苯作溶剂、碳酸氢钠作缚酸剂时, 室温反应4~5 h, 基本反应完全.所以最终选用该方法制备目标化合物7.

  合成中间体6时, 反应温度控制在125 ℃左右, 低温造成反应时间延长, 高温则会使2-溴丙酸甲酯回流, 影响反应的进行, 高温低温均会使产率降低.

  以化合物7c为例, 进行核磁氢谱和碳谱分析. CONH由于受到羰基和苯环的吸电子作用, 氢原子共振峰移向低场, δ值为9.88, 呈单峰.苯环上的氢的δ值在7.00~7.60左右, 且为多重峰, 位置较难归属. COCH₂CO中氢受到两个羰基的吸电子作用, δ值为3.08, 并且两个氢之间自偶合, 形成dd峰, 偶合常数为17.7 Hz. δ在172.5, 170.1, 163.6处的峰为三个羰基碳原子的信号峰.

  合成中间体4的方法主要有两种: (1) 中间体3和二氯亚砜回流制得; (2) 中间体3和草酰氯反应制得.第一种方法中氯化亚砜有强烈的刺激性气味, 并且毒性较大, 反应条件需要高温回流, 而第二种方法仅需要在冰浴条件下滴加草酰氯, 并加入催化量的DMF, 约2~3 h便可反应完全, 因此我们选择第二种方法来制得中间体4.

  1.2    杀菌活性

  表1 目标化合物的抑菌活性数据(5.0×10^-5 g/mL) Table1. Fungicidal activity of target compounds (5.0×10^-5 g/mL)

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  化合物	辣椒疫霉	番茄晚疫	水稻纹枯	棉花立枯	棉花枯萎	油菜菌核	番茄灰霉	番茄早疫
  7a	55.6	83.0	31.7	60.9	31.6	57.4	62.4	30.1
  7b	72.0	58.1	51.9	66.0	34.8	51.6	81.0	53.9
  7c	61.4	76.0	15.4	66.0	34.8	36.7	57.9	34.2
  7d	76.0	63.1	94.2	85.9	25.1	57.4	68.1	84.3
  7e	74.0	63.1	81.8	72.9	43.8	74.0	75.0	56.4
  7f	45.4	83.0	46.2	52.7	57.2	46.0	56.3	33.3
  7g	76.0	41.5	51.9	56.6	25.1	30.0	64.4	48.6
  7h	29.9	44.5	40.2	66.0	40.8	33.3	62.4	24.7
  7i	41.5	38.3	31.7	63.5	28.4	26.7	71.2	27.9
  7j	71.3	63.1	50.1	56.6	21.7	57.4	86.2	48.6
  7k	87.1	71.2	80.6	88.9	21.7	74.0	68.1	58.8
  7l	74.6	60.6	67.1	77.0	31.6	62.5	82.4	48.6
  7m	49.3	38.5	78.2	75.0	21.7	62.5	66.3	61.2
  7n	43.	28.9	59.0	72.9	18.3	33.3	58.4	48.6
  苯霜灵	74.6	90.2	74.0	60.7	80.9	40.8	36.7	75.0
  代谢物A	50.3	70.2	12.9	37.3	31.6	33.3	66.3	11.0

  表1 目标化合物的抑菌活性数据(5.0×10^-5 g/mL)
  Table1. Fungicidal activity of target compounds (5.0×10^-5 g/mL)

  将目标化合物配制成浓度为5.0×10^-5 g/mL药液, 采用菌体生长速率法^[20]对辣椒疫霉病菌(Phytophthora capsici)、番茄早疫病菌(Alternaria solani)、水稻纹枯病菌(Rhizoctonia solani)、油菜菌核病菌(Sclerotinia sclerotiorum)、番茄晚疫病菌(Phtophthora infestans)棉花立枯病菌(Rhizoctonia solani Kuhn)、番茄灰霉病菌(Botrytis cinerea Pers)、棉花枯萎病菌(Fsarium oxysporum)等8种菌进行了初步杀菌活性测试, 化合物杀菌普筛结果见表 1.

  由表 1可知, 目标化合物对8种菌均表现出一定的抑制活性, 部分化合物的抑菌活性优于苯霜灵.对棉花枯萎、油菜菌核、番茄晚疫和番茄早疫病原菌活性较差, 只有少数目标化合物表现出70%以上的抑菌活性.对水稻纹枯、棉花立枯、辣椒疫霉和番茄灰霉病原菌表现出良好抑菌活性, 其中, 化合物7k对辣椒疫霉病原菌的抑菌活性达到87%, 优于苯霜灵; 化合物7d对水稻纹枯和棉花立枯病原菌的抑制率分别为94.2%和85.9%, 化合物7k对水稻纹枯和棉花立枯病原菌的抑制率分别为80.6%和88.9%, 化合物7j对番茄灰霉病原菌的抑制率为86.2%.总体来说, 当取代基为吸电子基时, 化合物表现出更好的抑菌活性, 如化合物7d, 7k对大多数病原菌表现出良好的抑菌活性, 化合物7l和7m总体抑菌活性优于化合物7j;单吸电子基取代的化合物比双吸电子基取代的化合物具有更好的抑菌活性, 比如, 化合物7e抑菌活性优于化合物7a和7n.

  2    结论

  本文以苯霜灵代谢物N²-(丙二酸单酰基)-N¹-(2, 6-二甲苯基)丙氨酸甲酯结构为先导, 设计合成了14个结构新颖的双酰胺类化合物, 并对其进行了杀菌活性测试.生测结果表明, 目标化合物均有一定的杀菌活性, 其中, 化合物7d对番茄早疫、水稻纹枯和棉花立枯病原菌的抑制率分别为84.3%, 94.2%和85.9%, 均优于苯霜灵.苯环上R基团吸/推电子的性质和位置的不同, 对目标化合物杀菌活性有一定影响, 通过对目标化合物结构和抑菌活性之间的关系分析, 当取代基为单取代吸电子基时, 目标化合物的杀菌活性更好, 如化合物7d, 7e, 7k均表现出良好的离体杀菌活性, 其活体杀菌活性研究正在开展之中.

  3    实验部分

  3.1    仪器与试剂

  质谱采用Bruker Daltonics Bio-TOF-Q Ⅲ型质谱仪(ESIMS)测定; 核磁共振波谱使用Bruker Avance 300型核磁共振仪测定, 四甲基硅烷(TMS)为内标.所有试剂为国产市售分析纯, 无水溶剂按标准方法处理.

  3.2    实验方法

  3.2.2    中间体3的合成

  3-羰基-3-(4-甲基苯胺基)丙酸(3j)^[22]:白色固体, 产率91%. m.p. 119~120 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.59 (s, 1H, COOH), 10.03 (s, 1H, CONH), 7.49 (d, J=8.4 Hz, 2H, ArH), 7.12 (d, J=8.2 Hz, 2H, ArH), 3.36 (s, 2H, COC H₂CO), 2.26 (s, 3H, ArCH₃).

  3-羰基-3-(苯胺基)丙酸(3c)^[29]:棕色固体, 产率95%. m.p. 122~123 ℃(文献值163~165 ℃); ¹H NMR (300 MHz, DMSO-d₆)δ: 10.15 (s, 1H, CONH), 7.73~7.42 (m, 2H, ArH), 7.42~7.16 (m, 2H, ArH), 7.16~6.90 (m, 1H, ArH), 3.34 (s, 2H, COCH₂CO).

  3-羰基-3-(2-氯-4-三氟甲基苯胺基)丙酸(3b):白色固体, 产率86%. m.p. 133~134 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.84 (s, 1H, COOH), 10.09 (s, 1H, CONH), 8.21 (d, J=8.6 Hz, 1H, ArH), 7.91 (d, J=1.4 Hz, 1H, ArH), 7.71 (dd, J=8.7, 1.6 Hz, 1H, ArH), 3.55 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, DMSO-d₆) δ: 169.89, 165.72, 138.74, 126.68 (q, J=3.9 Hz), 125.82 (q, J=32.7 Hz), 124.82 (q, J=3.5 Hz), 123.61 (q, J=270.3 Hz), 124.16, 43.23; HRMS (ESI) calcd for C₁₀H₈ClF₃NO₃ [M+H]⁺: 282.0139, found 282.0142.

  3-羰基-3-(4-乙氧基苯胺基)丙酸(3g)^[31]:白色固体, 产率90%. m.p. 144~145 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 10.03 (s, 1H, COOH), 8.16~7.29 (m, 2H, ArH), 7.04~6.76 (m, 2H, ArH), 4.01 (q, J=6.9 Hz, 2H, ArOCH₂), 3.35 (s, 2H, COCH₂CO), 1.34 (t, J=7.0 Hz, 3H, CH₂CH₃).

  3-羰基-3-(4-溴-3-三氟甲基苯胺基)丙酸(3n):白色固体, 产率83%. m.p. 149~150 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.82 (s, 1H, COOH), 10.67 (s, 1H, CONH), 8.24 (d, J=2.2 Hz, 1H, ArH), 7.93~7.73 (m, 2H, ArH), 3.49 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, DMSO-d₆) δ: 169.13, 165.52, 139.04, 135.75, 128.90 (q, J=30.7 Hz), 124.09, 123.02 (q, J=271.5 Hz), 118.25 (q, J=5.8 Hz), 111.89, 44.31; HRMS (ESI) calcd for C₁₀H₈BrF₃NO₃[M+H]⁺: 325.9634, found 325.9632.

  3-羰基-3-(5-氟-2-甲基苯胺基)丙酸(3l):白色固体, 产率82%. m.p. 132~133 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.73 (s, 1H, COOH), 9.58 (s, 1H, CONH), 7.49 (dd, J=11.1 Hz, 2.7 Hz, 1H, ArH), 7.35~7.08 (m, 1H, ArH), 7.11~6.68 (m, 1H, ArH), 3.47 (s, 2H, COCH₂CO), 2.22 (s, 3H, ArCH₃); ¹³C NMR (75 MHz, DMSO-d₆) δ: 169.80, 165.17, 161.97 (d, J=238.3 Hz), 137.63 (d, J=10.8 Hz), 131.65 (d, J=9.0 Hz), 126.25 (d, J=2.9 Hz), 111.46 (d, J=20.8 Hz), 110.50 (d, J=24.9 Hz), 43.47, 17.27; HRMS (ESI) calcd for C₁₀H₁₁FNO₃ [M+H]⁺: 212.0717, found 212.0718.

  3-羰基-3-(2-氰基苯胺基)丙酸(3f)^[30]:浅黄色固体, 产率89%. m.p. 136~137 ℃ (文献值114~116 ℃); ¹H NMR (300 MHz, DMSO-d₆) δ: 12.68 (s, 1H, COOH), 10.37 (s, 1H, CONH), 7.81~7.65 (m, 3H, ArH), 7.36~7.30 (m, 1H, ArH), 3.48 (s, 2H, COCH₂CO).

  3-羰基-3-(2-甲氧基苯胺基)丙酸(3i)^[29]:白色固体, 产率86%. m.p. 147~148 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.71 (s, 1H, COOH), 9.57 (s, 1H, CONH), 8.11 (dd, J=8.0, 1.3 Hz, 1H, ArH), 7.15~7.01 (m, 2H, ArH), 6.98~6.87 (m, 1H, ArH), 3.84 (s, 3H, ArOCH₃), 3.54 (s, 2H, COCH₂CO).

  3-羰基-3-(4-甲氧基苯胺基)丙酸(3h)^[29]:无色晶体, 产率84%. m.p. 94~95 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.72 (s, 1H, COOH), 10.19 (s, 1H, CONH), 7.36 (t, J=2.2 Hz, 1H, ArH), 7.28 (t, J=8.1 Hz, 1H, ArH), 7.21~7.14 (m, 1H, ArH), 6.75~6.67 (m, 1H, ArH), 3.79 (s, 3H, OCH₃), 3.42 (s, 2H, COCH₂CO).

  3-羰基-3-(3-氯-4-甲基苯胺基)丙酸(3m)^[27]:黄色固体, 产率89%. m.p. 142~143 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.66 (s, 1H, COOH), 10, 21 (s, 1H, CONH), 7.78 (d, J=1.6 Hz, 1H, ArH), 7.38~7.17 (m, 2H, ArH), 3.34 (s, 2H, COCH₂CO), 2.24 (s, 3H, ArCH₃).

  3-羰基-3-(3-三氟甲基苯胺基)丙酸(3e)^[22]:白色固体, 产率86%. m.p. 104~105 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 12.72 (s, 1H, COOH), 10.47 (s, 1H, CONH), 8.09 (s, 1H, ArH), 7.75 (d, J=8.5 Hz, 1H, ArH), 7.53 (d, J=8.0 Hz, 1H, ArH), 7.45~7.21 (m, 1H, ArH), 3.41 (s, 2H, COCH₂CO).

  中间体3a的制备参考文献[28].取中间体2 (10 mmol), 四氢呋喃20 mL溶解, 后加入一水合氢氧化锂(0.84 g, 20 mmol)水溶液20 mL, 反应约1 h后, TLC检测反应完全, 加稀盐酸中和至酸性, 乙酸乙酯和水萃取, 有机相干燥后浓缩, 乙酸乙酯和石油醚重结晶得中间体3.

  3-羰基-3-(4-氯苯胺基)丙酸(3k)^[26]:白色固体, 产率87%. m.p. 145~146 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 1 2.64 (s, 1H, COOH), 10.24 (s, 1H, CONH), 7.70~7.54 (m, 2H, ArH), 7. 48~7.23 (m, 2H, ArH), 3.36 (s, 2H, COCH₂CO).

  3-羰基-3-(4-氯联苯-2-胺基)丙酸(3d):白色固体, 产率82%. m.p. 138~139 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 9.55 (s, 1H, CONH), 7.63 (d, J=7.9 Hz, 1H, ArH), 7.55~7.20 (m, 7H, ArH), 3.29 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, DMSO-d₆) δ: 169.92, 165.47, 137.80, 134.99, 134.83, 132.63, 131.27, 130.61, 128.85, 128.53, 126.60, 126.32, 43.34; HRMS (ESI) calcd for C₁₅H₁₂Cl-NO₃ [M+H]⁺: 290.0578, found 290.0375.

  3-羰基-3-(2, 5-二三氟甲基苯胺基)丙酸(3a):白色固体, 产率87%. m.p. 154~155 ℃; ¹H NMR (300 MHz, DMSO-d₆)δ: 10.18 (s, 1H, CONH), 8.08 (s, 1H, ArH), 7.98 (d, J=8.2 Hz, 1H, ArH), 7.78 (d, J=8.2 Hz, 1H, ArH), 3.50 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, DMSO-d₆) δ: 169.59, 166.07, 136.30, 133.17 (q, J=32.5 Hz), 128.01 (q, J=5.2 Hz), 126.42 (q, J=30.3 Hz), 124.68, 122.77 (q, J=3.4 Hz), 121.41 (q, J=271.2 Hz), 121.05 (q, J=272.2 Hz), 42.63; HRMS (ESI) calcd for C₁₁H₈F₆NO₃ [M+H]⁺: 316.0403, found 316.0406.

  3.2.1    中间体2的合成

  3-羰基-3-(4-溴-3-三氟甲基苯胺基)丙酸甲酯(2n):白色固体, 产率83%. m.p. 78~79 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.51 (s, 1H, CONH), 7.87 (d, J=2.3 Hz, 1H, ArH), 7.76~7.54 (m, J=15.8, 5.5 Hz, 2H, ArH), 3.82 (s, 3H, OCH₃), 3.50 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, DMSO-d₆) δ: 167.98, 164.99, 138.91, 135.83, 128.91 (q, J=30.7 Hz), 124.19, 123.02 (q, J=271.6 Hz), 118.31 (q, J=6.0 Hz), 112.07, 52.29, 43.77; HRMS (ESI) calcd for C₁₂H₁₆NO₄ [M+H]⁺: 339.9791, found 339.9792.

  3-羰基-3-(5-氟-2-甲基苯胺基)丙酸甲酯(2l):白色固体, 产率82%. m.p. 89~90 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.32 (s, 1H, CONH), 7.91 (dd, J=11.0, 2.7 Hz, 1H, ArH), 7.16~6.98 (m, 1H, ArH), 6.86~6.65 (m, 1H, ArH), 3.82 (s, 3H, OCH₃), 3.51 (s, 2H, COCH₂CO), 2.28 (s, 3H, ArCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 170.49, 162.65, 159.46 (d, J=239.6 Hz), 136.56 (d, J=11.2 Hz), 130.78 (d, J=9.0 Hz), 122.81 (d, J=3.0 Hz), 110.84 (d, J=21.3 Hz), 108.60 (d, J=26.8 Hz), 52.49, 40.74, 16.83; HRMS (ESI) calcd for C₁₁H₁₃FNO₃[M+H]⁺: 226.0874, found 226.0876.

  中间体2a~2n的制备参考文献[21].在250 mL的圆底烧瓶中加入化合物1 (20 mmol), 无水二氯甲烷30 mL溶解, 在冰浴和氮气保护条件下, 加入三乙胺(5.06 g, 50 mmol), 5 min后缓慢滴加丙二酸单甲酯酰氯二氯甲烷溶液15 mL (3.28 g, 24 mmol), 滴完后继续反应约3 h, 薄层色谱(TLC)监测反应完全, 甲醇淬灭反应.依次用1 mol/L盐酸、饱和氯化钠溶液、水洗涤有机相, 无水硫酸钠干燥后蒸去二氯甲烷, 柱层析纯化得到中间体2.

  3-羰基-3-(2, 5-二三氟甲基苯胺基)丙酸甲酯(2a):白色固体, 产率94%. m.p. 101~102 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.89 (s, 1H, CONH), 8.65 (s, 1H, ArH), 7.77 (d, J=8.2 Hz, 1H, ArH), 7.50 (d, J=8.2 Hz, 1H, ArH), 3.84 (s, 3H, OCH₃), 3.57 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, CDCl₃) δ: 169.34, 163.29, 135.40, 134.35 (q, J=33.3 Hz), 126.45 (q, J=5.4 Hz), 124.57 (q, J=271.6 Hz), 124.52 (q, J=272.0 Hz), 122.89 (q, J=30.7 Hz), 120.83 (q, J=5.3 Hz), 52.27, 40.66; HRMS (ESI) calcd for C₁₁H₈F₆NO₃ [M+H]⁺: 316.0403, found 316.0399.

  3-羰基-3-(2-氰基苯胺基)丙酸甲酯(2f)^[23]:淡黄色固体, 产率86%. m.p. 98~99 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.79 (s, 1H, CONH), 8.32 (d, J=8.3 Hz, 1H, ArH), 7.78~7.38 (m, 2H, ArH), 7.22~7.02 (m, 1H, ArH), 3.81 (s, 3H, OCH ₃), 3.57 (s, 2H, COCH₂CO).

  3-羰基-3-(2-甲氧基苯胺基)丙酸甲酯(2i)^[24]:无色晶体, 产率87%. m.p. 76~77 ℃; ¹H NMR (300 MHz, CDCl3) δ: 9.33 (s, 1H, CONH), 8.32 (dd, J=8.0, 1.6 Hz, 1H, ArH), 7.08~6.96 (m, 1H, ArH), 6.96~6.77 (m, 2H, ArH), 3.85 (s, 3H, OCH₃), 3.74 (s, 3H, OCH₃), 3.47 (s, 2H, COCH₂CO).

  3-羰基-3-(4-氯联苯-2-胺基)丙酸甲酯(2d):淡黄色固体, 产率83%. m.p. 87~88 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 8.74 (s, 1H, CONH), 8.20 (d, J=8.2 Hz, 1H, ArH), 7.68~7. 11 (m, 7H, ArH), 3.67 (s, 3H, OCH₃), 3.38 (s, 2H, COCH₂C O); ¹³C NMR (75 MHz, CDCl₃) δ: 169.04, 162.82, 136.43, 134.21, 133.76, 132.20, 130.56, 129.95, 128.83, 128.44, 124.85, 122.44, 52.30, 41.73; HRMS (ESI) calcd for C₁₆H₁₅ClNO₃[M+H]⁺: 304.0735, found 304.0741.

  3-羰基-3-(4-甲氧基苯胺基)丙酸甲酯(2h):淡棕色液体, 产率86%. ¹H NMR (300 MHz, CDCl₃) δ: 9.25 (s, 1H, CONH), 7.22 (t, J=2.2 Hz, 1H, ArH), 7.11 (t, J=8.1 Hz, 1H, ArH), 7.02~6.94 (m, 1H, ArH), 6.77~6.36 (m, 1H, ArH), 3.6 6 (s, 3H, OCH₃), 3.65 (s, 3H, OCH₃), 3.41 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, CDCl₃) δ: 169.11, 163.76, 159.86, 138.6 3, 129.41, 112.2, 110.19, 105.8, 54.98, 52.28, 42.25; HRMS (ESI) calcd for C₁₁H₁₄NO₄ [M+H]⁺: 224.0917, found 224.09 21.

  3-羰基-3-(4-乙氧基苯胺基)丙酸甲酯(2g):白色固体, 产率84%. m.p. 109~110 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.03 (s, 1H, CONH), 7.61~7.37 (m, 2H, ArH), 7.94~7.64 (m, 2H, ArH), 4.01 (q, J=7.0 Hz, 2H, ArOCH₂), 3.79 (s, 3H, OCH ₃), 3.47 (s, 2H, COCH₂CO), 1.40 (t, J=7.0 Hz, 3H, CH₂CH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 169.98, 162.58, 155. 80, 130.3 0, 121.73, 114.56, 63.49, 52.36, 41.28, 14.61. HRMS (ESI) calcd for C₁₂H₁₆NO₄ [M+H]⁺: 238.1074, found 238. 1073.

  3-羰基-3-(3-三氟甲基苯胺基)丙酸甲酯(2e)^[22]:白色固体, 产率89%. m.p. 63~64 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.46 (s, 1H, CONH), 7.87 (s, 1H, ArH), 7.75 (d, J=7.9 Hz, 1H, ArH), 7.55~7.30 (m, 2H, ArH), 3.81 (s, 3H, OCH₃), 3.52 (s, 2H, COCH₂CO).

  3-羰基-3-(2-氯-4-三氟甲基苯胺基)丙酸甲酯(2b):白色固体, 产率92%. m.p. 73~74 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 10.03 (s, 1H, CONH), 8.56 (d, J=8.7 Hz, 1H, ArH), 7.64 (s, 1H, ArH), 7.51 (d, J=8.7 Hz, 1H, ArH), 3.84 (s, 3H, O CH₃), 3.58 (s, 2H, COCH₂CO); ¹³C NMR (75 MHz, CDCl₃) δ: 169.75, 163.11, 137.41, 126.44 (q, J=33.4 Hz), 126.08 (q, J=3.9 Hz), 124.55 (q, J=3.7 Hz), 123.08 (q, J=270.3 Hz), 122.95, 121.08, 52.61, 41.15; HRMS (ESI) calcd for C₁₁H₁₀ClF₃NO₃ [M+H]⁺: 296.0296, found 296.0293.

  3-羰基-3-(4-氯苯胺基)丙酸甲酯(2k)^[26]:无色晶体, 产率89%. m.p. 115~116 ℃(文献值106~108 ℃); ¹H NMR (300 MHz, CDCl₃) δ: 9.30 (s, 1H, CONH), 7.52~7.43 (m, 2H, ArH), 7.29~7.22 (m, 2H, ArH), 3.77 (s, 3H, OCH₃), 3.48 (s, 2H, COCH₂CO).

  3-羰基-3-(3-氯-4-甲基苯胺基)丙酸甲酯(2m)^[27]:黄色固体, 产率88%. m.p. 93~94 ℃; ¹H NMR (300 MHz, DMSO-d₆) δ: 10.27 (s, 1H, CONH), 7.76 (d, J=2.0 Hz, 1H, ArH), 7.37~7.22 (m, 2H, ArH), 3.64 (s, 3H, OCH₃), 3.46 (s, 2H, COCH₂CO), 2.25 (s, 3H, ArCH₃).

  3-羰基-3-(苯胺基)丙酸甲酯(2c)^[22]:棕色固体, 产率96%. m.p. 43~44 ℃(文献值42~44 ℃); ¹H NMR (300 MHz, CDCl₃) δ: 9.34 (s, 1H, CONH), 7.61~7.47 (m, 2H, ArH), 7. 33~7. 18 (m, 2H, ArH), 7.12~6.95 (m, 1H, ArH), 3.67 (s, 3H, OCH₃), 3.45 (s, 2H, COCH₂CO).

  3-羰基-3-(4-甲基苯胺基)丙酸甲酯(2j)^[25]:无色晶体, 产率92%. m.p. 88~89 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.13 (s, 1H, CONH), 7.49~7.41 (m, 2H, ArH), 7.19~7.10 (m, 2H, ArH), 3.81 (s, 3H, OCH₃), 3.49 (s, 2H, COCH₂CO), 2.34 (s, 3 H, ArCH₃₎.

  3.3.3    N-(2, 6-二甲基苯基)丙酸甲酯(6)的合成

  中间体6的制备参考文献[33].在250 mL圆底烧瓶中加入2, 6-二甲基苯胺(1.21 g, 10 mmol)、碳酸氢钠(1.00 g, 12 mmol)、2-溴丙酸甲酯(5.01 g, 30 mmol), 122 ℃反应过夜, TLC检测反应完全, 过滤, 浓缩后柱层析分析分离得1.96 g棕色液体6^[32], 产率95%. ¹H NMR (300 MHz, DMSO-d₆) δ: 7.02 (d, J=7.4 Hz, 1H, ArH), 6.85 (t, J=7.4 Hz, 2H, ArH), 4.04 (q, J=7.0 Hz, 1H, COCH), 3.83 (s, 1H, NH), 3.72 (s, 3H, OCH₃), 2.36 (s, 6H, 2×ArMe), 1.44 (d, J=6.9 Hz, 3H, CHCH₃).

  3.3.4    目标化合物7的合成

  目标化合物7的制备参考文献[17]. 250 mL圆底烧瓶中, 加入中间体3 (10 mmol), 干燥二氯甲烷20 mL, 3滴DMF, 在冰浴和氮气保护条件下, 滴加草酰氯(1.27 g, 10 mmol), 滴加完毕后, 室温反应2~3 h, 得中间体4; 在冰浴和氮气保护条件下, 加入碳酸氢钠(1.26 g, 15 mmol), 将中间体4滴加至化合物6 (2.07 g, 10 mmol)的甲苯溶液中, 4 h后, TLC检测反应完全, 甲醇终止反应, 浓缩除去甲苯, 二氯甲烷和水萃取, 有机相干燥, 浓缩后柱层析得目标化合物7.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(5-氟-2-甲基苯胺基)丙酰胺基]丙酸甲酯(7l):黄色固体, 产率69%. m.p. 73~74 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 10.02 (s, 1H, CONH), 7.79 (dd, J=11.1, 2.7 Hz, 1H, ArH), 6.96~7.19 (m, 4H, ArH), 6.58~6.64 (m, 1H, ArH), 4.45 (q, J=7.4 Hz, 1H, CHCO), 3.70 (s, 3H, COOCH₃), 3.00 (dd, J=17.6, 24.5 Hz, 2H, COCH₂CO), 2.34, 2, 20, 2.05 (3s, 9H, 3×ArMe), 0.98 (d, J=7.5 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.10, 169.93, 163.41, 160.98 (d, J=240.1 Hz), 137.92, 137.01 (d, J=11.1 Hz), 136.68, 135.98, 130.65 (d, J=9.0 Hz), 129.43, 129.17, 128.79, 12 2.90 (d, J=3.2 Hz), 110.37 (d, J=21.3 Hz), 108.31 (d, J=26.8 Hz), 55.63, 51.97, 40.11, 18.28, 17.93, 16.93, 14.73; HRMS (ESI) calcd for C₂₂H₂₆FN₂O₄ (M+H)⁺: 401.1871, found 401.1874.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(3-氯-4-甲基苯胺基)丙酰胺基]丙酸甲酯(7m):黄色固体, 产率78%. m.p. 105~106 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.91 (s, 1H, CONH), 7.67 (d, J=2.1 Hz, 1H, ArH), 7.30 (d, J=2.6 Hz, 1H, ArH), 7.0 8~7.27 (m, 4H, ArH), 4.54 (q, J=7.4 Hz, 1H, CHCO), 3.81 (s, 3H, COOCH₃), 3.08 (dd, J=17.5, 29.6 Hz, 2H, COCH₂CO), 2.43, 2. 29, 2.15 (3s, 9H, 3×ArMe), 1.07 (d, J=7.5 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.29, 169.78, 163.54, 137.95, 136.75, 136.47, 136.04, 134.02, 131.36, 130.59, 129.43, 129.19, 128.82, 120.39, 118.15, 55.66, 52.17, 40.47, 19.20, 18.33, 18.06, 14.81; HRMS (ESI) calcd for C₂₂H₂₅ClN₂O₄ (M+H)⁺: 417.1576, found 417.1576.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(4-氯联苯-2-胺基)丙酰胺基]丙酸甲酯(7d):橘黄色固体, 产率70%. m.p. 76~77 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.77 (s, 1H, CONH), 8.12 (d, J=7.9 Hz, 1H, ArH), 7.42~7.46 (m, 2H, ArH), 7.32~7.38 (m, 3H, ArH), 7.08~7.27 (m, 5H, ArH), 4.40 (q, J=7.4 Hz, 1H, CHCO), 3.79 (s, 3H, COOCH₃), 2.98 (dd, J=17.7, 30.7 Hz, 2H, COCH₂CO), 2.40, 2.07 (2s, 6H, 2×ArMe), 1.02 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.16, 169.25, 16 3.84, 138.08, 136.72, 136.07, 134.57, 133.41, 132.72, 130.67, 130.04, 129.59, 129.09, 128.72, 128.65, 128.26, 124.70, 12 2.74, 55.47, 52.05, 40.15, 18.32, 17.99, 14.82; HRMS (ESI) calcd for C₂₇H₂₈ClN₂O₄ (M+H)⁺: 479.1732, found 479.1733.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(苯胺基)丙酰胺基]丙酸甲酯(7c):棕色固体, 产率92%. m.p. 98~99 ℃; ¹HNMR (300 MHz, CDCl₃) δ: 9.87 (s, 1H, CONH), 7.54 (d, J=7.6 Hz, 2H, ArH), 7.25~7.32 (m, 2H, ArH), 7.05~7.22 (m, 4H, ArH), 4.51 (q, J=7.4Hz, 1H, CHCO), 3.82 (s, 3H, COOCH₃), 3.06 (dd, J=17.7, 28.8 Hz, 2H, COCH₂CO), 2.14, 2.43 (2s, 6H, 2×ArMe), 1.05 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.52, 170.07, 163.64, 138.16, 137.76, 136.91, 136.24, 129.65, 129.36, 129.01, 128.84, 124.25, 120.20, 55.85, 52.40, 40.55, 18.52, 18.24, 15.01; HRMS (ESI) calcd for C₂₁H₂₅N₂O₄(M+H)⁺: 369.1809, found 369.1812.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(4-乙氧基苯胺基)丙酰胺基]丙酸甲酯(7g):黄色液体, 产率75%. ¹H NMR (300 MHz, CDCl₃) δ: 9.74 (s, 1H, CONH), 7.59~7.35 (m, 2H, ArH), 7.30~7.01 (m, 3H, ArH), 6.92~6.73 (m, 2H, ArH), 4.52 (q, J=7.5 Hz, 1H, CHCO), 4.10~3.90 (m, 2H, ArOCH₂), 3.83 (s, 3H, COOCH₃), 3.06 (dd, J=17.9, 28.5 Hz, 2H, COCH₂CO), 2.43, 2.14 (2s, 6H, 2×ArMe), 1.39 (t, J=7.0 Hz, 3H, CH₂CH₃), 1.07 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.23, 169.81, 163.07, 155.36, 137.84, 136.59, 135.94, 130.53, 129.29, 128.99, 128.65, 121.51, 114.30, 63.27, 55.48, 52.0 4, 40.10, 18.18, 17.91, 14.68, 14.49; HRMS (ESI) calcd for C₂₃H₂₉N₂O₅ (M+H)⁺: 413.2071, found 413.2076.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(4-溴-3-三氟甲基苯胺基)丙酰胺基]丙酸甲酯(7n):黄色液体, 产率74%. ¹H NMR (300 MHz, CDCl₃) δ: 10.22 (s, 1H, CONH), 7.89 (d, J=2.5 Hz, 1H, ArH), 7.66 (dd, J=8.7, 2.5 Hz, 1H, ArH), 7.55 (d, J=8.7 Hz, 1H, ArH), 7.12~7.28 (m, 3H, ArH), 4.55 (q, J=7.4 Hz, 1H, CHCO), 3.81 (s, 3H, COOCH₃), 3.11 (dd, J=17.6, 29.5 Hz, 2H, COCH₂CO), 2.44, 2.16 (2s, 6H, 2×ArMe), 1.07 (d, J=7.5 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.28, 169. 70, 164.01, 137.92, 137.16, 136.68, 135.90, 135.02, 129.51, 129.31, 128.88, 123.79, 119.08 (q, J=5.7 Hz), 55.70, 52.23, 40.46, 18.31, 18.04, 14.78; HRMS (ESI) calcd for C₂₂H₂₃BrF₃N₂O₄ (M+H)⁺: 515.0788, found 515.0792.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(2-氰基苯胺基)丙酰胺基]丙酸甲酯(7f):淡棕色固体, 产率85%. m.p. 92~93 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 10.72 (s, 1H, CONH), 8.21 (d, J=8.3 Hz, 1H, ArH), 7.61~7.41 (m, 2H, ArH), 7.26~6.96 (m, 4H, ArH), 4.53 (q, J=7.4Hz, 1H, CHCO), 3.82 (s, 3H, COOCH₃), 3.08 (dd, J=17.7, 28.7 Hz, 2H, COCH₂CO), 2.43, 2.14 (2s, 6H, 2×ArMe), 1.07 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ:172.21, 169.25, 164.27, 140.21, 138.06, 136.80, 135.79, 133.51, 132.45, 129.44, 129.19, 128.78, 124.15, 121.72, 115.89, 103.47, 55.61, 52.23, 39.96, 18.35, 18.03, 14.75; HRMS (ESI) calcd for C₂₂H₂₄N₃O₄ (M+H)⁺: 394.1761, found 394.1761.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(3-甲氧基苯胺基)丙酰胺基]丙酸甲酯(7h):黄色固体, 产率83%. m.p. 77~78 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.85 (s, 1H, CONH), 7.27 (t, J=2.2 Hz, 1H, ArH), 7.2 4~7.06 (m, 5H, ArH), 6.61~6.64 (m, 1 H, ArH), 4.53 (q, J=7.4 Hz, 1H, CHCO), 3.80 (s, 3H, COOCH₃), 3.76 (s, 3H, ArOCH₃), 3.09 (dd, J=17.4, 29.3 Hz, 2H, COCH₂CO), 2.43, 2.14 (2s, 6H, 2×ArMe), 1.07 (d, J=7.5 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.2 9, 169.78, 163.46, 159.84, 138.82, 137.96, 136.78, 136.12, 129.41, 129.29, 129.15, 128.81, 112.18, 109.99, 105.54, 55.66, 55.03, 52.15, 40.61, 18.32, 18.04, 14.81; HRMS (ESI) calcd for C₂₂H₂₇N₂O₅ (M+H)⁺: 399.1914, found 399.1918.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(3-三氟甲基苯基)丙酰胺基]丙酸甲酯(7e):黄绿色液体, 产率82%. ¹H NMR (300 MHz, CDCl₃) δ: 9.75 (s, 1H, CONH), 7.42~7.47 (m, 2H, ArH), 7.10~7.24 (m, 3H, ArH), 6.79~6.85 (m, 2H, ArH), 4.53 (q, J=7.4 Hz, 1H, CHCO), 3.82 (s, 3H, COOCH₃), 3.08 (dd, J=17.7, 28.7 Hz, 2H, COCH₂CO), 2.43, 2.14 (2s, 6H, 2×ArMe), 1.07 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.18, 169.60, 163.75, 138.01, 137.79, 136.56, 135.80, 13 0.81 (q, J=32.3 Hz), 129.33, 129.11, 128.99, 128.71, 123.56 (q, J=270.8 Hz), 122.75, 120.34 (q, J=3.8 Hz), 116.43 (q, J=4.0 Hz), 55.54, 52.07, 40.27, 18.15, 17.88, 14.62; HRMS (ESI) calcd for C₂₂H₂₄F₃N₂O₄(M+H)⁺: 437.1683, found 437.1685.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(2-氯-4-三氟甲基苯胺基)丙酰胺基]丙酸甲酯(7b):白色固体, 产率83%. m.p. 121~122 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 10.80 (s, 1H, CONH), 8.51 (d, J=8.6 Hz, 1H, ArH), 7.63 (d, J=1.8 Hz, 1H, ArH), 7.47 (dd, J=1.8 Hz, 8.8 Hz, 1H, ArH), 7.12~7.32 (m, 3H, ArH), 4.57 (q, J=7.4 Hz, 1H, CHCO), 3.81 (s, 3H, COOCH₃), 3.15 (dd, J=17.9, 25.3 Hz, 2H, COCH₂CO), 2.46, 2.16 (2s, 6H, 2×ArMe), 1.08 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.05, 16 9.29, 164.12, 137.98, 137.87, 136.73, 135.92, 129.45, 129.19, 128.78, 126.02 (q, J=33.3Hz), 126.01 (q, J=3.9 Hz), 124.29 (q, J=3.7 Hz), 123.23, 123.17 (q, J=270.2 Hz), 121.16, 55.64, 52.03, 40.34, 18.28, 17.92, 14.70; HRMS (ESI) calcd for C₂₂H₂₃ClF₃N₂O₄(M+H)⁺: 471.1293, found 471.1293.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(4-氯苯胺基)丙酰胺基]丙酸甲酯(7k):棕色液体, 产率71%. ¹H NMR (300 MHz, CDCl₃) δ: 10.00 (s, 1H, CONH), 7.49~7.54 (m, 2H, ArH), 7.12~7.29 (m, 5H, ArH), 4.54 (q, J=7.4 Hz, 1H, CHCO), 3.83 (s, 3H, COOCH₃), 3.08 (dd, J=17.9, 28.7 Hz, 2H, COCH₂CO), 2.43, 2.14 (2s, 6H, 2×ArMe), 1.08 (d, J=7.5 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.28, 169.69, 163.71, 137.94, 136.77, 136.33, 136.04, 129.40, 129.18, 128.80, 128.74, 128.53, 121.05, 55.62, 52.13, 40.64, 18.32, 18.05, 14.81; HRMS (ESI) calcd for C₂₁H₂₄ClN₂O₄ (M+H)⁺: 403.1419, found 403.1418.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(2-甲氧基苯胺基)丙酰胺基]丙酸甲酯(7i):棕色液体, 产率80%. ¹H NMR (300 MHz, CDCl₃) δ: 10.00 (s, 1H, CONH), 8.26 (dd, J=7.9, 1.6 Hz, 1H, ArH), 7.08~7.27 (m, 3H, ArH), 6.98~7.04 (m, 1H, ArH), 6.84~6.92 (m, 2H, ArH), 4.53 (q, J=7.4 Hz, 1H, CHCO), 3.87 (s, 3H, OCH₃), 3.78 (s, 3H, OCH₃), 3.09 (dd, J=17.1, 28.1 Hz, 2H, COCH₂CO), 2.42, 2.13 (2s, 6H, 2×ArMe), 1.05 (d, J=7.5 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ: 172.28, 169.43, 163.25, 148.57, 138.08, 136.93, 136.33, 129.34, 129.04, 128.74, 127.48, 123.76, 120.60, 120.12, 110.07, 55.66, 55.62, 51.94, 41.12, 18.37, 18.04, 14.88; HRMS (ESI) calcd for C₂₂H₂₇N₂O₅ (M+H)⁺: 399.1914, found 399.1913.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(2, 5-二三氟甲基苯胺基)丙酰胺基]丙酸甲酯(7a):淡棕色固体, 产率75%. m.p. 89~90 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 10.71 (s, 1H, CONH), 8.56 (s, 1H, ArH), 7.75 (d, J=8.3Hz, 1H, ArH), 7.45 (d, J=8.2 Hz, 1H, ArH), 7.37~7.06 (m, 3H, ArH), 4.55 (q, J=7.5 Hz, 1H, CHCO), 3.81 (s, 3H, COOCH₃), 3.11 (dd, J=18.1, 27.2 Hz, 2H, COCH₂CO), 2.45, 2.16 (2s, 6H, 2×ArMe), 1.07 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl3) δ: 171.81, 169.05, 164.16, 137.75, 136.58, 135.81, 135.70, 134.11 (q, J=33.0 Hz), 129.18, 128.98, 128.55, 126.39 (q, J=5.3 Hz), 122.91 (q, J=30.8 Hz), 120.97 (q, J=271.2 Hz), 120.88 (q, J=271.7 Hz), 120.52 (q, J=18.3 Hz), 120.32 (q, J=3.2 Hz), 55.49, 51.60, 39.72, 17.96, 17.51, 14.35; HRMS (ESI) calcd for C₂₃H₂₃F₆N₂ (M+H)⁺: 505.1557, found 505.1556.

  2-[N-(2, 6-二甲基苯基)-3-羰基-3-(4-甲基苯胺基)丙酰胺基]丙酸甲酯(7j):黄色固体, 产率76%. m.p. 103~104 ℃; ¹H NMR (300 MHz, CDCl₃) δ: 9.82 (s, 1H, CONH), 7.45 (d, J=8.4 Hz, 2H, ArH), 7.1 0~7.28 (m, 5H, ArH), 4.55 (q, J=7.4 Hz, 1H, CHCO), 3.84 (s, 3H, COOCH₃), 3.09 (dd, J=17.8, 29.0 Hz, 2H, COCH₂CO), 2.44, 2.31, 2.15 (3s, 9H, 3×ArMe), 1.09 (d, J=7.4 Hz, 3H, CHCH₃); ¹³C NMR (75 MHz, CDCl₃) δ:172.36, 169.96, 163.28, 138.00, 136.75, 136.11, 135.05, 133.62, 129.47, 129.16, 128.82, 120.08, 55.68, 52.20, 40.35, 20.67, 18.34, 18.06, 14.83; HRMS (ESI) calcd for C₂₂H₂₇N₂O₄ (M+H)⁺: 383.1965, found 383.1970.

  辅助材料(Supporting Information)    化合物2a~2n, 3a~3n, 7a~7n的¹H NMR谱图和化合物2a、2b、2d、2g、2h、2l、2n、3a、3b、3d、3g、3l、3n, 7a~7n的¹³C NMR谱图和HRMS.这些材料可以免费从本刊网站(http://sioc-journal.cn/)上下载.

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• 

  图 1  农药及其代谢产物

  Figure 1  Pesticides and their metabolites

  下载: 全尺寸图片 幻灯片
  

  图 2  设计目标化合物

  Figure 2  Target compound design

  下载: 全尺寸图片 幻灯片
  

  图式 1  目标化合物合成路线

  Scheme 1  Synthetic route of target compound

  7a: R=2, 5-(CF₃)₂-C₆H₃; 7b: R=2-Cl-4-CF₃-C₆H₃; 7c: R=C₆H₅; 7d: R=2-(4-Cl-C₆H₅)-C₆H₄; 7e: R=3-CF₃-C₆H₄; 7f: R=2-CN-C₆H₄; 7g: R=4-C₂H₅O-C₆H₄; 7h: R=3-CH₃O-C₆H₄; 7i: R=2-CH₃O-C₆H₄; 7j: R=4-Me-C₆H₄; 7k: R=4-Cl-C₆H₄; 7l: R=2-Me-5-F-C₆H₃; 7m: R=4-Me-3-Cl-C₆H₃; 7n: R=3-CF₃-4-Br-C₆H₃

  下载: 全尺寸图片 幻灯片

  表 1  目标化合物的抑菌活性数据(5.0×10^-5 g/mL)

  Table 1.  Fungicidal activity of target compounds (5.0×10^-5 g/mL)

  化合物	辣椒疫霉	番茄晚疫	水稻纹枯	棉花立枯	棉花枯萎	油菜菌核	番茄灰霉	番茄早疫
  7a	55.6	83.0	31.7	60.9	31.6	57.4	62.4	30.1
  7b	72.0	58.1	51.9	66.0	34.8	51.6	81.0	53.9
  7c	61.4	76.0	15.4	66.0	34.8	36.7	57.9	34.2
  7d	76.0	63.1	94.2	85.9	25.1	57.4	68.1	84.3
  7e	74.0	63.1	81.8	72.9	43.8	74.0	75.0	56.4
  7f	45.4	83.0	46.2	52.7	57.2	46.0	56.3	33.3
  7g	76.0	41.5	51.9	56.6	25.1	30.0	64.4	48.6
  7h	29.9	44.5	40.2	66.0	40.8	33.3	62.4	24.7
  7i	41.5	38.3	31.7	63.5	28.4	26.7	71.2	27.9
  7j	71.3	63.1	50.1	56.6	21.7	57.4	86.2	48.6
  7k	87.1	71.2	80.6	88.9	21.7	74.0	68.1	58.8
  7l	74.6	60.6	67.1	77.0	31.6	62.5	82.4	48.6
  7m	49.3	38.5	78.2	75.0	21.7	62.5	66.3	61.2
  7n	43.	28.9	59.0	72.9	18.3	33.3	58.4	48.6
  苯霜灵	74.6	90.2	74.0	60.7	80.9	40.8	36.7	75.0
  代谢物A	50.3	70.2	12.9	37.3	31.6	33.3	66.3	11.0

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