新型三唑哌嗪類衍生物的合成及生物活性

鄧珅釧, 李旭軍, 朱 祥, 杜曉英,2, 李俊凱,2, 徐志紅*,,2

(1. 長江大學 農學院，湖北 荊州 434025；2. 長江大學 農藥研究所，湖北 荊州 434025)

三唑類殺菌劑具有殺菌活性高、殺菌譜廣、內吸傳導性強和持效期長久等特點[1-3]，有些三唑類衍生物還具有殺蟲、除草或植物生長調節等活性[4-7]。隨著這類殺菌劑的廣泛應用，對其結構修飾的研究也更加深入。楊雙花等[8]設計合成的含醚鍵和α,β-不飽和酮結構的三唑類化合物對于小麥赤霉病、蘋果輪紋病具有良好的防效。杜琳等[9]通過酰胺化及疊氮-炔烴點擊反應合成的含三唑結構的環己甲酰胺衍生物，在50 mg/L 時對辣椒疫霉病的最高抑制率可達91%。童建穎等[10]通過活性亞結構拼接的方法引入鄰氟苯甲酸，合成的系列鄰氟苯基三唑類衍生物對黃瓜真菌類病害具有較好的防效。三唑類衍生物的構效關系表明，三唑環是三唑類殺菌劑的關鍵藥效基團，2,4-二氟苯基是重要的藥效基團，羥基 (酮基) 對于抗真菌發揮一定作用[11-13]。

近年來，在醫藥領域哌嗪及其衍生物作為一種重要的中間體，為治療腫瘤、糖尿病、真菌類和精神類疾病的新藥研究上提供了新的選擇[14-17]。在新農藥的創制中，劉長令等[18-19]合成含有哌嗪的二鹵丙烯醚類衍生物，對昆蟲的卵、幼蟲和成蟲均表現出較高的殺蟲活性；吳清來等[20]以天然產物Aspernigerin 為先導化合物，設計合成的Aspernigerin類似物，具有一定的殺菌、殺蟲和除草活性。

基于本課題組前期的研究結果[21]，本研究在保留1,2,4-三唑環和2,4-二氟苯基的三唑衍生物的基礎上進行結構改造，將哌嗪活性亞結構進行拼接，合成了新型三唑哌嗪類衍生物，并進行了殺菌活性測定。鑒于含哌嗪基團的衍生物還具有一定的殺蟲活性[22-24]，本研究同時進行了殺蟲活性測定。

目標化合物的合成路線如圖式1 所示。

1 材料與方法

1.1 儀器與藥劑

所有化學藥品均為市售國產試劑。97% 1,2,4-三唑，上海朋索生化科技有限公司；98% 2-氯-2,4-二氟苯乙酮，上海笛柏生物科技有限公司；無水哌嗪，上海阿拉丁生化科技股份有限公司；96%三唑酮 (triadimefon) 原藥，山東濰坊潤豐化工股份有限公司；97%三唑醇 (triadimenol) 原藥，江蘇七洲綠色化工股份有限公司；96% 氰氟蟲腙(metaflumizone) 原藥，京博農化科技有限公司。

Bruker Avance DPX400 型核磁共振儀 (以TMS 作內標物，CDCl3作溶劑)，德國-瑞士布魯克光譜公司；WRR 熔點測定儀 (熔點未校正)，上海精密科學儀器有限公司；Themo scientific QExactive 型高分辨質譜儀 (ESI-MS)，賽默飛世爾科技 (中國) 有限公司；Digital water bath SB-1000 旋轉蒸發儀，日本EYELA 公司。

1.2 供試真菌及蟲源

水稻紋枯病菌Rhizoctonia solani、小麥赤霉病菌Fusarium graminearum、番茄早疫病菌Altemaria solani、西瓜枯萎病菌Fusarium oxysporum、白芨白絹病菌Sclerotium rolfii 和水稻稻瘟病菌Magnaporthe oryzae，均由長江大學農學院植物病理實驗室提供。

斜紋夜蛾Spodoptera litura 為大小一致、健康的3 齡幼蟲，由長江大學昆蟲研究所提供。

1.3 化合物的合成

1.3.1 2,4-二氟-2-(1H-1,2,4-三唑-1-基) 苯乙酮的合成 (2) 參考路小娟等[25]的方法合成，得到白色固體 (2)，收率58%，熔點99.5～101.2 ℃ (文獻值[25]：收率41.7%，m.p. 103～105 ℃)。

1.3.2 1-(2,4-二氟苯基)-1-羥基-2-(1H-1,2,4-三唑-1-基)-乙醇的合成 (3) 參考Nelson 等[26]的方法合成，得到淡黃色固體，收率93%，熔點118～120 ℃(文獻值[26]：收率85%，m.p. 118～120 ℃)。

1.3.3 1-[(取代芐基) 甲基]哌嗪的通用合成方法(5) 參考韓生華等[27]的方法合成，略有改進。向250 mL 圓底燒瓶中加入無水哌嗪8.27 g (0.096 mol)和K2CO34.42 g (0.032 mol) 溶于100 mL 正丁醇中的溶液，冰浴條件下攪拌溶解；用恒壓滴定漏斗向其中滴加相應的氯芐 (n(無水哌嗪) : n (氯芐) =6 : 1) 與正丁醇的混合溶液，加熱回流3 h，薄層層析 (TLC，V(石油醚) : V(乙酸乙酯) = 4 : 1) 監測反應進程。反應完畢，減壓濃縮，加入水溶解，用濃鹽酸調節pH 值為3～4。用乙酸乙酯 (30 mL ×3) 萃取，無水硫酸鈉干燥，旋干，再用甲醇溶解，減壓濃縮至干。得到白色固體1-[(取代芐基)甲基]哌嗪純品，收率80～90%(文獻值[27]：收率73.5%～81.8%)。

1.3.4 2-氯-1-{4-[(取代芐基) 甲基]哌嗪-1-基}-1-乙酮的合成 (6) 參考文獻[28]的方法合成。向250 mL 單口圓底燒瓶中加入相應的1-[(取代芐基)甲基]哌嗪 (5) 和二氯甲烷50 mL，常溫下攪拌至溶解；隨后加入K2CO31.11 g (0.008 mol)，冰浴下攪拌15 min；將0.91 g (0.008 mol) 氯乙酰氯(n(1-[(取代芐基) 甲基]哌嗪) : n(氯乙酰氯) = 1 : 3)快速溶于20 mL 二氯甲烷中并用恒壓滴定漏斗緩慢滴加到單口圓底燒瓶中，滴加完后室溫反應1 h。過濾，減壓濃縮，再用甲醇溶解，減壓濃縮至干，分別得到淡黃色油狀液體或黃色油狀液體或褐色油狀液體產物6a-6t，其中化合物6g、6h、6i、6o 和6r 為未知化合物。

1.3.5 1-(4-(取代芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮化合物(7a～7t) 的合成通法 向250 mL 單口圓底燒瓶中加入化合物3 2.15 g (0.01 mol) 和二氯甲烷50 mL，攪拌溶解后加入叔丁醇鈉1.54 g (0.016 mol)，充分攪拌20 min。緩慢滴加化合物6 2.22 g (0.008 mol)溶于50 mL 二氯甲烷的溶液，在室溫下反應6 h，TLC (V(石油醚) : V(乙酸乙酯) = 3 : 1) 監測反應進程。待反應液澄清后，過濾，濾液濃縮至干，用柱層析分離純化 (V(石油醚) : V(乙酸乙酯) = 1 : 1)，無水硫酸鈉干燥后得產物 (7a～7t)，收率53%～76%。

7a：1-(4-(2-甲基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體, 收率68%.1H NMR (400 MHz, CDCl3), δ: 8.21 (s, 1H), 7.90 (s,1H), 7.35 – 7.26 (m, 1H), 7.24 – 7.11 (m, 4H), 6.93 – 6.82 (m,2H), 5.19 – 5.13 (m, 1H), 4.49 (dd, J = 10.4, J = 6.0 Hz, 2H),4.07 (d, J = 13.6 Hz, 1H), 3.95 (d, J = 13.6 Hz, 1H), 3.52 (s,2H), 3.44 (s, 2H), 3.23 – 3.09 (m, 2H), 2.39 (s, 2H), 2.35 (s,3H), 2.31 (t, J = 4.8 Hz, 2H).13C NMR (101 MHz, CDCl3), δ:166.42, 163.31 (dd, JC-F= 250.5, JC-F= 12.1 Hz), 160.83 (dd,JC-F= 249.3, JC-F= 12.1 Hz), 151.94, 144.54, 137.75, 135.79,130.58, 130.04, 127.54, 125.74, 120.18 (dd, J = 13.7, J = 3.8 Hz), 112.28 (dd, J = 21.3, J = 3.6 Hz), 104.55 (t, J = 25.4 Hz),74.37, 68.06, 60.87, 54.15, 53.14, 52.70, 44.98, 42.04, 19.39.HRMS: 計算值C24H27F2N5O2([M+H]+) 456.220 6, 實測值456.2207.

7b：1-(4-(3-甲基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率68%.1H NMR (400 MHz, CDCl3), δ: 8.23 (s, 1H), 7.89 (s,1H), 7.32 (dd, J = 14.8, J =8.4 Hz, 1H), 7.20 (t, J = 7.6 Hz,1H), 7.08 (dd, J = 11.2, J =6.4 Hz, 3H), 6.91– 6.81(m, 2H),5.16 (dd, J = 7.2, J = 4.4 Hz, 1H), 4.57 – 4.43 (m, 2H), 4.08 (dd, J =10.0, J = 6.4 Hz, 1H), 3.95 (d, J = 13.6 Hz, 1H), 3.54 (s, 2H),3.45 (s, 2H), 3.27 – 3.12 (m, 2H), 2.37 (s, 2H), 2.33 (s, 3H),2.31 (s, 2H).13C NMR (101 MHz, CDCl3), δ: 166.34, 163.17(dd, JC-F= 243.5, JC-F= 7.2 Hz), 160.64 (dd, JC-F= 242.6,JC-F= 7.2 Hz), 151.68, 144.44, 138.00, 137.08, 129.25 (dd,J = 9.8, J =5.5 Hz), 128.20, 126.30, 120.03 (dd, J = 13.7, J =3.7 Hz),112.13 (dd, J = 21.3, J = 3.6 Hz), 104.38(t,J = 25.4 Hz), 74.19, 67.82, 62.76, 53.95, 52.83, 52.38, 44.63,41.68, 21.40. HRMS: 計算值C24H27F2N5O2([M+H]+), 456.220 6,實測值456.2206.

7c：1-(4-(4-甲基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1，2，4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體, 收率66%.1H NMR (400 MHz, CDCl3), δ: 8.20 (s, 1H), 7.90 (s,1H), 7.33 – 7.27 (m, 1H), 7.18 (d, J = 8.0 Hz, 2H), 7.13 (d, J =8.0 Hz, 2H), 6.92 – 6.81 (m, 2H), 5.15 (t, J = 5.6 Hz, 1H), 4.49(d, J = 5.6 Hz, 2H), 4.06 (d, J = 13.6 Hz, 1H), 3.94 (d, J = 13.6 Hz, 1H), 3.60 – 3.50 (m, 2H), 3.46 (s, 2H), 3.26 – 3.12 (m,2H), 2.41 – 2.36 (m, 2H), 2.34 (s, 3H), 2.31 (t, J = 5.2 Hz, 2H).13C NMR (101 MHz, CDCl3), δ: 166.40, 163.27 (dd, JC-F= 244.0,JC-F= 6.7 Hz), 160.73 (dd, JC-F= 243.1, JC-F=6.7 Hz), 151.81,144.50, 137.67, 135.69, 130.51, 129.98, 129.31 (dd, J = 9.8,J = 5.5 Hz),127.47, 125.67, 120.12 (dd, J = 13.7, J = 3.7 Hz),112.21 (dd, J = 21.3, J = 3.6 Hz), 104.47(t, J = 25.4 Hz),74.28, 67.92, 60.77, 54.05, 53.04, 52.62, 44.88, 41.96, 19.32.HRMS: 計算值C24H27F2N5O2([M+H]+) 456.220 6, 實測值456.2208.

7d：1-(4-(2-氟芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率68%.1H NMR (400 MHz, CDCl3), δ: 8.19 (d, J = 2.4 Hz, 1H),7.91 (s, 1H), 7.36 – 7.28 (m, 2H), 7.28 – 7.24 (m, 1H), 7.14 –7.00 (m, 2H), 6.92 – 6.81 (m, 2H), 5.16 (dd, J = 7.2, J =4.4 Hz,1H), 4.55 – 4.48 (m, 2H), 4.06 (d, J = 13.6 Hz, 1H), 3.94 (d,J = 13.6 Hz, 1H), 3.58 (s, 2H), 3.55 (s, 2H), 3.26 – 3.13 (m,2H), 2.43 (s, 2H), 2.36 (s, 2H).13C NMR (101 MHz, CDCl3),δ: 166.47, 163.66 (dd, JC-F= 212.4, JC-F=38.8 Hz), 161.96 (d,JC-F= 247.5 Hz), 160.53 (dd, JC-F= 159.7, JC-F= 90.5 Hz),151.04, 144.34, 144.71, 131.88 (d, J = 4.4 Hz), 129.51 (dd, J = 11.7,J = 3.5 Hz), 124.31 (d, J = 3.6 Hz), 124.05 (t, J = 8.0 Hz),120.26 (dd, J = 13.7, J = 3.7 Hz), 115.72 (d, J = 22.2 Hz),112.45 (dd, J = 21.3, J = 3.6 Hz), 104.70 (t, J = 25.5 Hz),74.52, 68.13, 62.92, 54.26, 52.86, 52.44, 44.94, 42.01. HRMS:計算值C23H24F3N5O2([M+H]+) 460.195 5, 實測值460.1956.

7e：1-(4-(3-氟芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率65%.11H NMR (400 MHz, CDCl3), δ: 8.21 (s, 1H), 7.91 (s,1H), 7.33 (dd, J = 8.4, J = 6.4 Hz, 1H), 7.28 – 7.25 (m, 1H),7.05 (dd, J = 10.4, J = 4.4 Hz, 2H), 6.98 –6.82(m, 3H), 5.16(dd, J = 7.6, J = 4.4 Hz, 1H), 4.55 – 4.45 (m, 2H), 4.07 (d, J =13.6 Hz, 1H), 3.95 (d, J = 13.6 Hz, 1H), 3.56 (s, 2H), 3.48 (s,2H), 3.26 – 3.14 (m, 2H), 2.39 (t, J = 4.4 Hz, 2H), 2.32 (t, J =4.0 Hz, 2H).13C NMR (101 MHz, CDCl3), δ: 166.48, 163.90(dd, JC-F= 133.4, JC-F=117.3 Hz), 162.60 (d, JC-F=247.4 Hz),160.77 (dd, JC-F= 238.4, JC-F= 11.5 Hz), 151.93, 144.55,140.43 (d, J = 7.0 Hz), 129.97 (d, J = 8.2 Hz), 129.36 (dd, J =9.8, J = 5.5 Hz), 124.63 (d, J = 2.8 Hz), 120.11 (dd, J = 13.6, J = 3.8 Hz),115.81 (d, J = 21.3 Hz), 114.41 (d, J = 21.1 Hz), 112.30 (dd,J = 21.4, J = 3.6 Hz), 104.56 (t, J = 25.5 Hz), 74.37, 68.02,62.30, 54.12, 53.02, 52.58, 44.85, 41.89. HRMS: 計算值C23H24F3N5O2([M+H]+) 460.195 5, 實測值460.1958.

7f：1-(4-(4-氟芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率62%.1H NMR (400 MHz, CDCl3), δ: 8.20 (s, 1H), 7.90 (s,1H), 7.34 – 7.29 (m, 1H), 7.26 (dd, J = 8.4, J = 5.6 Hz, 2H),7.04 – 6.97 (m, 2H), 6.93 – 6.84 (m, 2H),, 5.16 (t, J = 5.6 Hz,1H), 4.49 (dd, J = 10.4, J = 5.6 Hz, 2H), 4.07 (d, J = 13.6 Hz,1H), 3.94 (d, J = 13.6 Hz, 1H), 3.60 – 3.50 (m, 2H), 3.46 (s,2H), 3.24 – 3.13 (m, 2H), 2.37 (t, J = 4.4 Hz, 2H), 2.30 (t, J =4.8 Hz, 2H).13C NMR (101 MHz, CDCl3), δ: 166.38, 163.65(dd, JC-F= 175.2, JC-F= 75.4 Hz), 162.14 (d, JC-F= 245.4 Hz), 160.53(dd, JC-F= 195.7, JC-F= 53.2 Hz), 151.92, 144.49, 133.25 (d,J = 2.9 Hz), 130.69 (d, J = 7.9 Hz), 129.34 (dd, J = 9.9, J = 5.5 Hz),120.12 (dd, J = 13.7, J = 3.8 Hz), 115.32 (d, J = 21.3 Hz),112.24 (dd, J = 21.3, J = 3.6 Hz), 104.52 (t, J = 25.4 Hz),74.36, 68.07, 62.06, 54.10, 52.94, 52.46, 44.82, 41.82. HRMS:計算值C23H24F3N5O2([M+H]+) 460.195 5, 實測值460.1958.

7g：2-((4-(2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基)乙氧基) 乙酰基) 哌嗪-1-基) 甲基) 苯甲腈，黃色油狀液體,收率63%.1H NMR (400 MHz, CDCl3), δ: 8.24 (s, 1H),7.91 (s, 1H), 7.66 (d, J = 7.6Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H),7.51 (d, J = 7.6 Hz, 1H), 7.39 (t, J = 7.2 Hz, 1H), 7.32 (t, J =7.4 Hz, 1H), 6.95 – 6.82 (m, 2H), 5.17 (dd, J = 7.2, J =4.4 Hz,1H), 4.56 – 4.46 (m, 2H), 4.08 (d, J = 13.6 Hz, 1H), 3.96 (d, J =13.6 Hz, 1H), 3.69 (s, 2H), 3.55 (d, J = 4.8 Hz, 2H), 3.28 –3.14 (m, 2H), 2.49 – 2.43 (m, 2H), 2.41 (t, J = 4.4 Hz, 2H).13C NMR (101 MHz, CDCl3), δ: 166.36, 163.19 (dd, JC-F= 243.5,JC-F= 7.2 Hz), 160.66 (dd, JC-F= 242.6, JC-F=7.3 Hz), 151.72,144.44, 141.63, 133.15, 132.72, 130.11, 129.25 (dd,J = 9.8, J = 5.5 Hz), 127.97, 120.01 (dd, J = 13.7, J = 3.8 Hz),117.75, 113.12, 112.16 (dd, J = 21.3, J = 3.7 Hz), 104.41 (t, J =25.4 Hz), 74.21, 67.89, 60.31, 53.96, 52.83, 52.30, 44.66,41.67. HRMS: 計算值C24H24F2N6O2([M+H]+) 467.200 2, 實測值467.2003.

7h：4-((4-(2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基)乙氧基) 乙酰基) 哌嗪-1-基) 甲基) 苯甲腈，黃色油狀液體,收率65%.1H NMR (400 MHz, CDCl3), δ: 8.25 (s, 1H), 7.90(s, 1H), 7.62 (d, J = 8.4 Hz, 2H), 7.46 (d, J = 8.4 Hz, 2H), 7.34(dd, J = 14.4, J = 7.8 Hz, 1H), 6.94 – 6.82 (m, 2H), 5.19 (dd,J = 7.2, J = 4.4 Hz, 1H), 4.56 – 4.46 (m, 2H), 4.10 (d, J =13.6 Hz, 1H), 3.97 (d, J = 13.6 Hz, 1H), 3.56 (s, 4H), 3.30 –3.15 (m, 2H), 2.39 (s, 2H), 2.34 (t, J = 4.8Hz, 2H).13C NMR(101 MHz, CDCl3), δ: 166.23, 163.02 (dd, JC-F= 241.8,JC-F=8.8 Hz), 160.50 (dd, JC-F= 241.1, JC-F= 8.8 Hz), 151.57,144.30, 143.39, 132.06 (d, J = 8.8 Hz), 129.36, 129.14 (dd, J =9.8, J = 5.5 Hz), 119.91, 118.74, 111.99 (dd, J = 21.3, J = 3.6 Hz),110.94, 104.24 (t, J = 25.5 Hz), 74.04, 67.80, 61.98, 53.80,52.84, 52.37, 44.55, 41.51. HRMS: 計算值C24H24F2N6O2([M+H]+) 467.200 2, 實測值467.2006.

7i：1-(4-(4-乙基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率75%.1H NMR (400 MHz, CDCl3), δ: 8.22 (s, 1H), 7.91 (s,1H), 7.30 (dd, J = 15.2, J = 8.4 Hz,1H), 7.21 (d, J = 8.4 Hz,2H), 6.93 – 6.81 (m, 4H), 5.19 – 5.12 (m, 1H), 4.54 – 4.44 (m,2H), 4.06 (d, J = 13.6 Hz, 1H), 3.98 – 3.89 (m, 1H), 3.80 (s,3H), 3.55 (s, 2H), 3.47 (s, 2H), 3.24 – 3.16 (m, 4H), 2.40 (s,2H), 2.33 (s, 2H).13C NMR (101 MHz, CDCl3), δ: 166.47,163.33 (d, JC-F= 239.0 Hz), 160.84 (d, JC-F= 119.18 Hz),159.20, 151.87, 144.55, 130.66, 129.38 (dd, J = 9.8, J = 5.5 Hz),128.91, 120.13, 113.93, 112.30 (dd, J = 21.3, J = 3.6 Hz),104.57 (t, J = 25.5 Hz), 74.39, 68.02, 62.20, 54.14, 52.78,52.31, 44.68, 41.72, 21.18. HRMS: 計算值C25H29F2N5O2([M+H]+) 470.236 4, 實測值470.2362.

7j：1-(4-(4-甲氧基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體,收率69%.1H NMR (400 MHz, CDCl3), δ: 8.36 – 8.12 (m,1H), 8.05 – 7.82 (m, 1H), 7.35 – 7.23 (m, 3H), 7.19 (d, J = 7.8 Hz, 2H), 6.92 – 6.82 (ddd, J = 21.6, J = 13.6, J = 5.6 Hz, 2H),4.56 – 4.41 (m, 2H), 4.13 – 4.02 (m, 2H), 3.93 (d, J = 13.6 Hz,1H), 3.67 (s, 3H), 3.35 (d, J = 3.6 Hz, 2H), 2.65 (m,J = 7.6 Hz, 2H), 2.56 (s, 2H), 2.52 (s, 2H).13C NMR (101 MHz,CDCl3), δ: 166.26, 163.15 (dd, JC-F= 251.0, JC-F= 12.0 Hz),160.66 (dd, JC-F= 250.1, JC-F= 12.0 Hz), 159.07, 151.77,144.39, 130.52, 129.20 (dd, J = 9.8, J = 5.5 Hz), 119.94 (dd,J = 13.7, J = 3.7 Hz), 113.79, 112.12 (dd, J = 21.4, J = 3.6 Hz), 104.40(t, J = 25.4 Hz), 74.24, 67.88, 62.01, 55.27, 53.95, 52.60,52.12, 44.45, 41.47. HRMS: 計算值C24H27F2N5O3([M+H]+) 472.215 4, 實測值472.2155.

7k：1-(4-(2-氯芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體, 收率76%.1H NMR (400 MHz, CDCl3), δ: 8.21 (s, 1H), 7.91 (s,1H), 7.42 (dd, J = 7.6, J = 1.8 Hz, 1H), 7.38 – 7.28 (m, 2H),7.27 – 7.17 (m, 2H), 6.93 – 6.81 (m, 2H), 4.54 – 4.47 (m, 2H),4.15 – 4.04 (m, 2H), 3.95 (d, J = 13.6 Hz, 1H), 3.67 – 3.51 (m,4H), 3.24 – 3.15 (m, J = 13.2, J = 8.4 Hz, 2H), 2.46 (t, J = 4.0 Hz,2H), 2.39 (t, J = 5.2 Hz, 2H).13C NMR (101 MHz, CDCl3), δ:166.27, 163.15 (d, JC-F= 238.6 Hz), 160.66 (dd, JC-F= 249.7,JC-F= 12.0 Hz), 151.79, 144.37, 135.13, 134.41, 130.74,129.59, 129.20 (dd, J = 9.9, J = 5.4 Hz), 128.47, 126.69,119.98 (dd, J = 13.7, J = 3.8 Hz), 112.12 (dd, J = 21.4, J = 3.6 Hz),104.40 (t, J = 25.4 Hz), 74.22, 67.91, 59.05, 53.97, 52.90,52.46, 44.75, 41.79. HRMS: 計算值C23H24ClF2N5O2([M+H]+) 476.165 9, 實測值476.1657.

7l：1-(4-(3-氯芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率73%.1H NMR (400 MHz, CDCl3), δ: 8.22 (s, 1H), 7.92 (s,1H), 7.36 – 7.29 (m, 2H), 7.25 (d, J = 1.2 Hz, 2H), 7.17 (t, J =3.2 Hz, 1H), 6.93 – 6.82 (m, 2H), 5.18 – 5.14 (m, 1H), 4.52 –4.48 (m, 2H), 4.07 (d, J = 13.6 Hz, 1H), 3.94 (d, J = 13.6 Hz,1H), 3.56 (s, 2H), 3.47 (s, 2H), 3.27 – 3.15 (m, 2H), 2.39 (t, J =4.4 Hz, 2H), 2.32 (t, J = 4.8 Hz, 2H).13C NMR (101 MHz,CDCl3), δ: 166.51, 162.16 (d, JC-F= 12.0 Hz), 162.10 (d, JC-F=12.0 Hz), 151.86, 144.55, 139.63, 134.51, 129.84, 129.39 (dd,J = 9.9, J = 5.5 Hz), 129.22, 127.79, 127.35, 120.10 (dd, J =13.6, J = 3.8 Hz),112.33 (dd, J = 21.4, J = 3.5 Hz), 104.60 (t, J = 25.5 Hz),74.41, 68.03, 62.24, 54.16, 52.97, 52.54, 44.79, 41.82. HRMS:計算值C23H24ClF2N5O2([M+H]+) 476.165 9, 實測值476.166 6.

7m：1-(4-(4-氯芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體, 收率74%.1H NMR (400 MHz, CDCl3), δ: 8.19 (s, 1H), 7.90 (s,1H), 7.35 – 7.27 (m, 3H), 7.23 (d, J = 8.4 Hz, 2H), 6.93 – 6.82(m, 2H), 5.16 (dd, J = 6.4, J = 4.8 Hz, 1H), 4.53 – 4.45 (m,2H), 4.06 (d, J = 13.6 Hz, 1H), 3.94 (d, J = 13.6 Hz, 1H), 3.57 –3.50 (m, 2H), 3.45 (s, 2H), 3.26 – 3.11 (m, 2H), 2.37 (t, J =4.8 Hz, 2H), 2.29 (t, J = 5.2 Hz, 2H).13C NMR (101 MHz,CDCl3), δ: 166.52, 163.30 (dd, JC-F= 251.1, JC-F= 12.0 Hz),160.81 (dd, JC-F= 250.2, JC-F= 12.0 Hz), 151.69, 144.53,135.80, 133.29, 130.58, 129.35 (dd, J = 9.8, J = 5.5 Hz),128.68, 120.03 (dd, J = 13.7, J = 3.7 Hz), 112.28 (dd, J = 21.3,J = 3.7 Hz), 104.54 (t, J = 25.4 Hz), 74.32, 67.92, 62.01, 54.10,52.84, 52.40, 44.69, 41.74. HRMS: 計算值C23H24ClF2N5O2([M+H]+) 476.165 9, 實測值476.1664.

7n：1-(4-(4-異丙基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體,收率65%.1H NMR (400 MHz, CDCl3), δ: 8.20 (s, 1H), 7.90(s, 1H), 7.35 – 7.28 (m, 1H), 7.23 – 7.14 (m, 4H), 6.92 – 6.80(m, 2H), 5.16 (t, J = 5.6 Hz, 1H), 4.53 – 4.47 (m, 2H), 4.06 (d,J = 13.6 Hz, 1H), 3.94 (d, J = 13.6 Hz, 1H), 3.59 – 3.51 (m,2H), 3.47 (d, J = 7.6 Hz, 2H), 3.24 – 3.12 (m, 2H), 2.93 – 2.88(m, 1H), 2.38 (t, J = 4.8 Hz, 2H), 2.31 (t, J = 5.0 Hz, 2H), 1.25(s, 3H), 1.24 (d, J = 1.6 Hz, 3H).13C NMR (101 MHz, CDCl3),δ: 166.60, 163.34 (dd, JC-F= 251.5, JC-F= 11.4 Hz), 160.85(dd, JC-F= 250.8, JC-F= 11.5 Hz), 151.67, 148.50, 144.57,133.72, 129.62, 129.39 (dd, J = 9.9, J = 5.4 Hz), 126.65,120.06 (d, J = 10.0 Hz), 112.32 (dd, J = 21.2, J = 3.6 Hz),104.58 (t, J = 25.5 Hz), 74.38, 67.94, 62.38, 54.15, 52.67,52.18, 44.52, 41.58, 33.97, 24.16, 21.20. HRMS: 計算值C26H31F2N5O2([M+H]+) 484.251 9, 實測值484.2522.

7o：1-(4-(2-硝基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體, 收率69%.1H NMR (400 MHz, CDCl3), δ: 8.20 (s, 1H), 7.91 (s,1H), 7.81 (d, J = 7.6 Hz, 1H), 7.57 – 7.51 (m, 2H), 7.46 – 7.39(m, 1H), 7.33 – 7.28 (m, 1H), 6.96 – 6.81 (m, 2H), 5.20 – 5.11(m, 1H), 4.49 (dd, J = 10.4, J =5.6 Hz, 2H), 4.06 (d, J = 13.6 Hz,1H), 3.93 (d, J = 13.6 Hz, 1H), 3.79 (s, 2H), 3.51 (t, J = 9.2 Hz, 2H), 3.21 – 3.09 (m, 2H), 2.36 (s, 2H), 2.32 (s, 2H).13C NMR (101 MHz, CDCl3), δ: 166.42, 163.30 (dd, JC-F= 250.6,JC-F= 12.0 Hz), 160.81 (dd, JC-F=249.6, JC-F= 12.0 Hz),151.91, 150.00, 144.50, 133.08, 132.59, 131.09, 129.36 (d, J =4.3 Hz), 128.48, 124.70, 120.05 (d, J = 3.8 Hz), 112.26 (dd,J = 21.4, J = 3.6 Hz), 104.54 (t, J = 25.4 Hz), 74.38, 68.04,59.03, 54.10, 53.16, 52.56, 44.83, 41.84. HRMS: 計算值C23H24F2N6O4([M+H]+) 487.190 0, 實測值487.1904.

7p：1-(4-(4-硝基芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率73%.1H NMR (400 MHz, CDCl3), δ: 8.20 (d, J = 3.2 Hz, 2H),8.18 (s, 1H), 7.91 (s, 1H), 7.51 (d, J = 8.8 Hz, 2H), 7.36-7.28(m, 1H), 6.93-6.83 (m, 2H), 5.17 (dd, J = 6.8, J = 4.8 Hz, 1H),4.53 – 4.45 (m, 2H), 4.07 (d, J = 13.6 Hz, 1H), 3.95 (d, J =13.6 Hz, 1H), 3.58 (d, J = 9.6 Hz, 4H), 3.27 – 3.14 (m, 2H),2.40 (t, J = 4.8 Hz, 2H), 2.33 (t, J = 4.8 Hz, 2H).13C NMR(101 MHz, CDCl3), δ: 166.45, 163.32 (dd, JC-F= 250.8, JC-F=12.0 Hz), 160.83 (dd, JC-F= 249.8, JC-F= 12.0 Hz), 151.92,147.47, 145.65, 144.50, 129.60, 129.36 (dd, J = 9.8, J = 5.5 Hz),123.81, 120.05 (dd, J = 13.7, J = 3.8 Hz), 112.29 (dd, J = 21.3,J = 3.6 Hz), 104.57 (t, J = 25.5 Hz), 74.39, 68.08, 62.00, 54.11,53.16, 52.69, 44.82, 41.80. HRMS: 計算值C23H24F2N6O4([M+H]+) 487.190 0, 實測值487.1903.

7q：2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基)-1-(4-(萘-2-基甲基) 哌嗪-1-基) 乙酮，褐色油狀液體, 收率53%.1H NMR (400 MHz, CDCl3), δ: 8.27 – 8.23 (m, 1H),8.21 (s, 1H), 7.90 (s, 1H), 7.85 (dd, J = 6.8, J = 2.4 Hz, 1H),7.79 (dd, J = 7.6, J = 1.6 Hz, 1H), 7.55 – 7.46 (m, 2H), 7.43 –7.38 (m, 2H), 7.30 (dd, J = 11.6, J = 5.0 Hz, 1H), 6.91 – 6.81(m, 2H), 5.16 (t, J = 5.6 Hz, 1H), 4.49 (d, J = 5.6 Hz, 2H), 4.07(d, J = 13.6 Hz, 1H), 3.96 – 3.88 (m, 3H), 3.53 (s, 2H), 3.17(dd, J = 10.4, J = 5.2 Hz, 2H), 2.46 (t, J = 4.4 Hz, 2H), 2.37 (t,J = 4.8 Hz, 2H).13C NMR (101 MHz, CDCl3), δ: 166.42,162.93 (m), 161.28 (m), 151.91, 144.54, 134.05, 133.35,132.61, 129.45, (d, J = 16.2 Hz), 128.57 (d, J = 16.2 Hz),127.78, 125.98 (d, J = 12.6 Hz), 125.92, 125.27, 124.83,120.17 (dd, J = 13.8, J = 3.8 Hz), 112.28 (dd, J = 21.4, J = 3.8 Hz),104.56 (t, J = 25.4 Hz), 74.37, 68.07, 61.19, 54.16, 53.23,52.86, 44.94, 42.01. HRMS:計算值C27H27F2N5O2([M+H]+)492.220 6, 實測值492.2208.

7r：1-(4-(4-叔丁基苯) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體, 收率67%.1H NMR (400 MHz, CDCl3), δ: 8.22 (s, 1H), 7.90 (s,1H), 7.34 (d, J = 8.2 Hz, 2H), 7.32 – 7.29 (m, 1H), 7.22 (d, J =8.2 Hz, 2H), 6.91 – 6.81 (m, 2H), 5.16 (dd, J = 6.6, J = 4.8 Hz, 1H),4.52 – 4.48 (m, 2H), 4.07 (d, J = 13.6 Hz, 1H), 3.95 (d, J =13.6 Hz, 1H), 3.58 – 3.51 (m, 2H), 3.47 (s, 2H), 3.25 –3.15 (m, 2H), 2.39 (s, 2H), 2.33 (s, 2H), 1.31 (s, 9H).13C NMR(101 MHz, CDCl3), δ: 166.32, 163.19 (dd, JC-F= 243.3, JC-F=7.3 Hz), 160.65 (dd, JC-F= 242.6, JC-F= 7.3 Hz), 151.72,150.33, 144.42, 134.10, 129.27 (dd, J = 9.8, J = 5.5 Hz),128.94, 125.29, 120.07 (dd, J = 13.7, J = 3.7 Hz), 112.13 (dd,J = 21.3, J = 3.6 Hz), 104.40 (t, J = 25.4 Hz), 74.23, 67.92,62.39, 53.99, 52.87, 52.33, 44.69, 41.70, 34.53, 31.41, 31.11.HRMS: 計算值C27H33F2N5O2([M+H]+) 498.267 5 實測值498.2678.

7s：1-(4-(2,4-二氯芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，黃色油狀液體,收率59%.1H NMR (400 MHz, CDCl3), δ: 8.22 (s, 1H), 7.91(s, 1H), 7.38 (dd, J = 8.6, J = 5.2 Hz, 2H), 7.35 – 7.27 (m, 1H), 7.23 (dd,J = 8.4, J = 2.2 Hz, 1H), 6.93 – 6.83 (m, 2H), 5.17 (dd, J = 6.8,J = 4.8 Hz, 1H), 4.55 – 4.47 (m, 2H), 4.08 (d, J = 13.6 Hz,1H), 3.96 (d, J = 13.6 Hz, 1H), 3.56 (s, 4H), 3.20 – 3.14 (m,2H), 2.46 – 2.33 (m, 4H).13C NMR (101 MHz, CDCl3),δ: 166.29, 163.18 (dd, JC-F= 243.4, JC-F=7.4 Hz), 160.65 (dd,JC-F= 242.5, JC-F= 7.4 Hz), 151.78, 144.40, 134.95, 133.92,133.43, 131.50, 129.32, 129.22 (m), 127.06, 120.02 (dd, J =13.7, J = 3.8 Hz), 112.13 (dd, J = 21.3, J = 3.6 Hz), 104.40 (t,J = 25.4 Hz), 74.23, 67.95, 58.46, 53.96, 52.89, 52.45, 44.74,41.74. HRMS: 計算值C23H23Cl2F2N5O2([M+H]+) 510.127 0,實測值510.1277.

7t：1-(4-(4-(三氟甲基) 芐基) 哌嗪-1-基)-2-(1-(2,4-二氟苯基)-2-(1H-1,2,4-三唑-1-基) 乙氧基) 乙酮，褐色油狀液體,收率63%.1H NMR (400 MHz, CDCl3), δ: 8.23 (s, 1H), 7.92(s, 1H), 7.58 (s, 1H), 7.52 (dd, J = 12.2, J = 7.6 Hz, 2H), 7.45(t, J = 7.6 Hz, 1H), 7.34 – 7.27 (m, 1H), 6.93 – 6.82 (m, 2H),5.16 (dd, J = 6.8, J = 4.8 Hz, 1H), 4.56 – 4.45 (m, 2H), 4.07 (d,J = 13.6 Hz, 1H), 3.95 (d, J = 13.6 Hz, 1H), 3.55 (d, J =6.4 Hz, 4H), 3.29 – 3.13 (m, 2H), 2.41 – 2.31 (m, 4H).13C NMR (101 MHz, CDCl3), δ: 166.50, 163.33 (dd, JC-F= 251.0,JC-F= 12.1 Hz), 160.84 (dd, JC-F= 250.0, JC-F= 12.0 Hz),151.83, 144.53, 138.71, 132.46, 130.91 (d, J = 32.2 Hz),129.36 (dd, J = 9.9, J = 5.5 Hz), 129.02, 125.77, 124.42 (q,JC-F= 3.9 Hz), 122.94, 120.07 (dd, J = 13.7, J = 3.8 Hz),112.29 (dd, J = 21.3, J = 3.6 Hz), 104.56 (t, J = 25.4 Hz),74.38, 68.02, 62.30, 54.13, 53.00, 52.57, 44.78, 41.81. HRMS:

計算值C24H24F5N5O2([M+H]+) 510.192 3, 實測值510.1931.

1.4 生物活性測定

1.4.1 殺菌活性的測定 采用菌絲生長速率法[29]測定化合物7a～7t 對6 種植物病原真菌的殺菌活性，藥劑濃度為200 μmol/L。將目標化合物溶解于1 mL 丙酮中，用0.1%的吐溫水溶液定容至50 mL 后加入熱的PDA 培養基中，制成含藥培養基。在含藥培養基上接入菌餅。每個藥劑3 次重復，以三唑酮為對照藥劑。置于生化培養箱 (溫度28 ℃) 中培養，待空白對照菌落長至接近2/3 培養皿直徑時，以十字交叉法測定各處理菌落的直徑，按 (1) 式計算供試化合物對病原菌的生長抑制率。

式中：I—菌絲生長抑制率，%；D0—空白對照菌落直徑，mm；Dt—藥劑處理菌落直徑, mm。

選擇抑制率大于60% 的化合物測定其EC50值。采用等倍稀釋法配制目標化合物的系列質量濃度為400、200、100、50 和25 μmol/L，采用上述菌絲生長速率法分別測定各濃度藥液對供試植物病原菌的抑制率。以三唑醇為對照藥劑，采用軟件SPSS 計算EC50值和95%置信限。

1.4.2 殺蟲活性的測定 參考FAO 推薦的觸殺法[30]。將目標化合物用丙酮配制成質量濃度為100 mg/L。選取大小一致的3 齡幼蟲，用微量點滴器點滴到斜紋夜蛾幼蟲前胸背板上，每頭給藥1 mL。將處理后幼蟲轉移至90 mm 培養皿中，常規條件下飼養。每處理重復3 次。以丙酮處理為空白對照，以氰氟蟲腙為藥劑對照。置于恒溫箱[(26 ± 1)℃，相對濕度65%，光周期12L：12D]內飼養48 h 后，觀察結果并計算死亡率。幼蟲的死亡標準[31]：蟲體干癟、變黑，以毛筆輕觸蟲體，無明顯自主反應者判斷為死亡。

2 結果與討論

2.1 化合物的合成

為了避免引入其他無機堿，本研究采用了6 倍量的哌嗪，以較高收率 (80%～90%) 獲得了單取代哌嗪中間體5。在制備中間體6 的過程中，為了便于產物的純化，使用了3 倍量的氯乙酰氯，而過量的氯乙酰氯經處理后生成酸，用飽和碳酸鈉溶液充分洗滌即可除去。

2.2 目標化合物的生物活性

表1 目標化合物對6 種植物病原菌菌絲生長的抑制作用Table1 Inhibitory effect of target compounds against six pathogenic fungi in vitro

2.2.1 殺菌活性 離體殺菌活性測試結果 (表1)表明：在200 μmol/L 時，絕大多數目標化合物對水稻紋枯病菌的生長均有不同程度的抑制作用 (抑制率大于40%)；化合物7n、7r 和7s 對小麥赤霉病菌的抑制率分別為6 0.1 9%、6 2.2 7% 和50.00%，其中7n 和7r 與對照藥劑三唑酮抑菌效果相當；同時部分化合物如7m、7p、7r 和7s 對水稻稻瘟病菌也有一定的殺菌活性；但大多數化合物對番茄早疫病菌、白芨白娟病菌和西瓜枯萎病菌的殺菌活性較弱。

根據初篩結果測定了殺菌活性較高化合物的EC50值。從表2 數據可以看出，化合物7r 對水稻紋枯病菌和小麥赤霉病菌的殺菌活性最強，其EC50值分別為49.49 μmol/L 和109.7 μmol/L，其中對小麥赤霉病菌的毒力高于對照藥劑三唑醇。

表2 部分目標化合物對供試病原菌的EC50 值Table2 The EC50 values of some target compounds against test phytopathogens

初步的結構-活性關系表明，在目標化合物的苯環上引入不同取代基 (R)，會對化合物的抑菌活性產生影響。從7n 和7r 對小麥赤霉病菌的抑制活性可以看出，當R 為異丙基和叔丁基時，其殺菌活性優于R 為甲基、乙基和其他取代基團時的活性，說明烷基基團越大，空間位阻越大，抑菌活性越強。對水稻紋枯病菌，化合物7s(R=2,4-2Cl) 和7l(R=3-Cl) 活性明顯優于7k(R=2-Cl) 和7m (R=4-Cl)，說明雙鹵素原子鄰對位取代和單原子間位取代的抑菌活性高于鄰對位單取代。

2.2.2 殺蟲活性 殺蟲活性測定結果表明：在100 mg/L 下，部分化合物處理斜紋夜蛾幼蟲24 h 后，試蟲出現行動遲緩、取食較少現象；處理48 h后，化合物7g、7j、7o 和7p 對斜紋夜蛾幼蟲表現出一定的殺蟲效果，其中7o 致死率最高，為40.00%，略低于對照藥劑氰氟蟲腙 (表3)。

表3 目標化合物對斜紋夜蛾的殺蟲活性Table3 Insecticidal activities of target compounds against Spodoptera litura

3 結論

本研究通過活性亞結構拼接的方式，設計合成了20 個未見文獻報道的新型三唑哌嗪類衍生物，其結構均通過1H NMR、13C NMR 和高分辨質譜表征確認。

殺菌活性結果表明：在200 μmol/L 時，大部分化合物對水稻紋枯病菌表現出中等偏上的抑制作用，但不及對照藥劑三唑酮；化合物7n 和7r 對小麥赤霉病菌表現出良好的抑制作用，高于對照藥劑三唑醇，說明哌嗪環的引入增強了這類結構對小麥赤霉病菌的抑制作用；同時部分化合物對水稻稻瘟病菌也有一定的抑制作用，但對番茄早疫病菌、白芨白娟病菌和西瓜枯萎病菌的抑制作用偏低，結果不如預期。殺蟲活性測定結果表明，化合物7o 對斜紋夜蛾幼蟲表現出一定的殺蟲活性，但略低于對照藥劑氰氟蟲腙，殺蟲活性并不理想。說明哌嗪基團的引入對于三唑類化合物的抑菌活性和殺蟲活性的改善作用有限，三唑類衍生物的結構改造有待進一步研究。