賈長青, 楊冬燕, 車傳亮, 馬永強, 芮昌輝, 閆曉靜, 覃兆海

(1. 中國農業(yè)大學理學院, 北京 100193; 2. 中國農業(yè)科學院植物保護研究所, 北京 100193)

?

1H-1,2,4-三唑-5-氨基甲酸酯類化合物的合成、 結構表征及殺蟲、 抑菌活性

賈長青1, 楊冬燕1, 車傳亮1, 馬永強1, 芮昌輝2, 閆曉靜2, 覃兆海1

(1. 中國農業(yè)大學理學院, 北京 100193; 2. 中國農業(yè)科學院植物保護研究所, 北京 100193)

摘要通過活性基團拼接的方法, 將氨基甲酸酯結構和1H-1,2,4-三唑-5-胺有機結合, 設計合成了一系列結構新穎的多取代1H-1,2,4-三唑-5-氨基甲酸酯類化合物(5和6), 所有化合物的結構經核磁共振(NMR), 紅外光譜(IR), 高分辨質譜(HRMS)及元素分析等方法確證.初步殺蟲活性測定結果表明, 大部分化合物對桃蚜[Myzus persicae(Sulzer)]具有良好的殺蟲活性, 致死率大于90%. 精密毒力測試結果表明, 化合物5b的半致死濃度(LC50)為9.49 μg/mL, 具有進一步研究開發(fā)的價值.抑菌活性初步測試結果表明該類化合物的抑菌活性一般, 在50 μg/mL濃度下少數化合物也表現出一定的離體抑菌活性. 對此類化合物的構效關系進行了討論.

關鍵詞1H-1,2,4-三唑-5-胺; 氨基甲酸酯; 殺蟲活性; 抑菌活性

氨基甲酸酯類農藥是基于天然產物毒扁豆生物堿開發(fā)的繼有機磷之后出現的一類高活性農藥品種, 作為殺蟲劑、 殺菌劑及除草劑被廣泛應用. 在該類農藥中, 氨基甲酸酚(或肟)酯多表現出強烈的殺蟲活性, 如呋喃丹、 涕滅威及克百威等, 而芳氨基甲酸酯則表現出良好的殺菌活性, 如多菌靈(1)和苯菌靈(2)等(結構見圖1). 雖然該類化合物發(fā)現較早并得到較充分的開發(fā), 但氨基甲酸酯類化合物表現出的高效、 速效及易降解等特點使氨基甲酸酯結構仍具有活力[1,2]. 如BASF公司將氨基甲酸酯結構引入到Strobilurin類殺菌劑中, 于1993年研發(fā)了唑菌胺酯(Pyraclostrobin, 3, 結構見圖1)[3,4]. 1,2,4-三唑類農藥是另一個重要的農藥品種, 該品種具有廣譜、 高效的抑菌活性和優(yōu)良的殺蟲、 除草及植物生長調節(jié)活性, 使得1,2,4-三唑及其衍生物受到廣泛關注[5～12]. 另外, 在醫(yī)藥領域中, 許多含有1,2,4-三唑氨基甲酸酯結構的化合物具有抗菌[13]、 抗腫瘤[14]及抑制脂肪酸酰胺水解酶(FAAH)[15]等活性; 在農藥領域中, 含有1,2,4-三唑基團的氨基甲酸酯化合物也表現出較好的殺螺[16]、 殺菌[17]和植物生長調節(jié)[18]活性.

Fig.1　Structrue of compounds 1—3

前文[19]報道了一類結構新穎且合成簡便的1,3-二取代-1H-1,2,4-三唑-5-氨類化合物(4), 其結構易于衍生且部分化合物具有較好的抑菌活性, 該類化合物具有作為農藥活性結構的潛力. 鑒于多取代1H-1,2,4-三唑-5-氨基甲酸酯類化合物的合成和生物活性研究尚未見報道, 也為了進一步尋找活性優(yōu)異的1H-1,2,4-三唑類衍生物, 本文通過活性基團拼接的方法, 將氨基甲酸酯結構單元引入到該類化合物中, 設計合成了目標化合物(如Scheme1所示), 并對其抑菌和殺蟲活性進行了研究.

4a, 5a: R1=n-Pr, R2=Cl; 4b, 5b: R1=n-Bu, R2=Cl; 4c, 5c: R1=H, R2=Et; 4d, 5d: R1=H, R2=Allyl; 4e, 5e: R1=2-NO2, R2=Et; 4f, 5f: R1=2-NO2, R2=n-Pr; 4g, 5g: R1=2-NO2, R2=n-Bu; 4h, 5h: R1=2-NO2, R2=Allyl; 4i, 5i, 6b: R1=2-Cl, R2=Et; 4j, 5j: R1=2-Cl, R2=n-Bu; 4k, 5k: R1=2-Cl, R2=Allyl; 4l, 5l: R1=4-OCH3, R2=Et; 4m, 5m: R1=4-OCH3, R2=n-Bu; 4n, 5n: R1=4-Cl, R2=Et; 4o, 5o: R1=4-Cl, R2=n-Pr; 4p, 5p: R1=4-Cl, R2=Allyl; 4q, 5q: R1=4-NO2, R2=Et; 4r, 5r: R1=4-NO2, R2=n-Pr; 4s, 5s: R1=4-NO2, R2= n-Bu; 4t, 5t: R1=4-F, R2=Et; 4u, 5u: R1=4-F, R2=n-Pr; 4v, 5v: R1=4-F, R2=Allyl; 4w, 5w: R1=4-F, R2=n-Bu; 4x, 5x: R1=4-NO2, R2=Allyl; 4y, 6a: R1=H, R2=n-Bu; 4z, 6c: R1=2-OCH3, R2=Et; 4aa, 6d: R1=2-OCH3, R2=Allyl; 4ab, 6e: R1=4-OCH3, R2=n-Pr; 4ac, 6f: R1=4-OCH3, R2=Allyl; 4ad, 6g: R1=4-Cl, R2=n-Bu. Scheme 1　Design and synthesis of target compounds 5 and 6

1實驗部分

1.1試劑與儀器

所用試劑均為國產分析純, 購自北京偶合科技有限公司.

Yanagimoto熔點測定儀(日本MFG公司);AvanceDPX300型核磁共振儀(TMS為內標,DMSO-d6為溶劑, 德國Bruker公司);ST-Carloerba元素分析儀(采用FlashEA1112法, 德國Elementar公司);VECTOR22型紅外光譜儀(KBr壓片, 德國Bruker公司);ThermoScientificLTQOrbitrapDiscovery型高分辨質譜儀(德國Bremen公司).

1.2實驗過程

1.2.1中間體化合物4的合成中間體化合物4參照文獻[19]方法合成, 其中化合物4a, 4b和4q～4x尚未見文獻報道, 各中間體的表征結果分別列于表1和表2.

Table 1　Melting point data of intermediate compounds 4

Table 2　1H NMR data of intermediate compounds 4a, 4b and 4q—4x

1.2.2目標化合物5a～5x的合成稱取中間體化合物4(1.5mmol, 0.5g)加入50mL單口燒瓶中, 用10mL丙酮使其完全溶解, 再加入1mL3mol/LNaOH水溶液, 室溫下攪拌反應0.5h后, 加入氯甲酸乙酯(1.6mol, 0.17g), 繼續(xù)反應2h. 過濾, 濾去析出的固體, 將濾液濃縮, 用硅膠柱層析[V(乙酸乙酯)∶V(石油醚)=1∶6], 得目標化合物5a～5x.

1.2.3目標化合物6a～6g的合成稱取中間體化合物4(1.5mmol, 0.5g)加入50mL單口燒瓶中, 用10mL丙酮使其完全溶解, 再加入1mL3mol/LNaOH水溶液, 室溫下攪拌反應30min后, 加入氯甲酸乙酯(3.2mol, 0.34g), 繼續(xù)反應4h. 過濾, 濾去析出的固體鹽, 將濾液濃縮, 用硅膠柱層析[V(乙酸乙酯)∶V(石油醚)=1∶8], 得目標化合物6a～6g. 目標化合物的表征結果見表3和表4.

Table 3　Appearance, melting point, yield, elemental analysis and IR data of target compounds

Continued.

Compd.Appearancem.p./℃Yield(%)Elementalanalysis(%,calcd.)CHNIR(KBr),ν/cm-15nWhitepowder114—1154452.79(52.98)5.11(5.13)18.88(19.01)3166,2977,1741,1580,1462,1430,1260,1233,1132,1089,837,7505oWhitepowder115—1164854.50(54.46)5.50(5.55)17.90(18.15)3448,3191,2970,2937,1777,1744,1464,1586,1426,1374,1324,1259,1288,1092,857,7515pWhitepowder80—815354.71(54.82)5.11(4.93)18.20(18.26)3468,3184,3079,2982,1743,1579,1557,1457,1428,1256,1230,1064,838,7515qYellowsolid197—1985651.17(51.14)4.98(4.95)22.90(22.94)3463,3209,2986,1699,1605,1568,1521,1341,1274,1237,1068,854,7275rYellowsolid169—1705452.61(52.66)5.46(5.37)21.77(21.93)3216,3083,2978,2944,1699,1604,1561,1523,1342,1265,1227,1065,854,7275sYellowsolid150—1514654.33(54.05)5.92(5.75)20.76(21.01)3237,3090,2960,2874,1704,1604,1561,1524,1468,1341,1263,1070,866,7275tWhitepowder114—1155356.26(56.11)5.59(5.43)20.08(20.13)3183,3069,2976,2941,1745,1601,1561,1466,1431,1262,1233,1058,845,7555uWhitepowder115—1164757.44(57.52)5.99(5.86)19.08(19.17)3473,3186,3066,2976,2941,1745,1601,1563,1467,1428,1255,1229,1150,1061,848,7575vWhitepowder89—904958.11(57.92)5.43(5.21)19.29(19.30)3186,3066,2980,2938,1744,1601,1562,1467,1431,1256,1228,1188,1062,848,7565waWhitepowder54—55483194,3067,2962,2932,2876,1742,1609,1567,1469,1431,1249,1226,1060,850,7555xbYellowsolid175—176463434,3210,2984,1701,1604,1653,1524,1464,1324,1206,1066,855,7276aWhitepowder83—844459.71(59.99)6.73(6.71)15.54(15.55)3453,2981,2960,2874,1778,1645,1589,1580,1447,1370,1323,1264,1208,1099,855,775,7026bWhitepowder96—974552.24(52.39)5.28(5.22)15.33(15.27)3445,3050,2988,1781,1657,1613,1582,1562,1471,1367,1324,1252,1201,1150,1073,857,773 6cWhitepowder93—944056.40(56.34)6.20(6.12)15.54(15.46)3438,2973,2939,2841,1774,1658,1601,1445,1372,1305,1264,1202,1075,850,7756dWhitepowder104—1054157.73(57.75)6.01(5.92)15.00(14.96)3440,2904,1775,1658,1603,1469,1437,1327,1215,1140,1045,927,7736eWhitesolid107—1084257.46(57.74)6.54(6.43)14.93(14.88)3443,2971,2935,2842,1776,1645,1615,1587,1562,1508,1372,1325,1259,1210,1180,1068,857,7366fWhitepowder98—994257.77(57.75)5.81(5.92)15.03(14.96)3442,2978,2840,1778,1646,1589,1563,1507,1372,1327,1264,1212,1179,1067,858,7646gcWhitepowder89—90493439,3091,2983,2937,2899,2875,1778,1647,1587,1442,1370,1321,1265,1204,1100,857,730

a.HRMScalcd.forC15H19FN4O2[M+H]+307.1564,found307.1564; b.HRMScalcd.forC14H15N5O4[M+H]+318.1197,found318.1201; c.HRMScalcd.forC18H23ClN4O4[M+H]+395.1480,found395.1484.

Table 4　1H NMR and 13C NMR data of target compounds

Continued.

Compd.1HNMR(DMSO-d6,300MHz),δ13CNMR(DMSO-d6,75MHz),δ5c10.08(brs,1H,NH),7.96—7.92(m,5H,Ph),4.16(q,J=7.08Hz,2H,OCH2),4.05(q,J=7.20Hz,2H,CH2N),1.38(t,J=7.20Hz,3H,OCH2CH3),1.24(t,J=7.11Hz,3H,CH3)158.76,153.96,147.11,131.14,129.18,128.82,125.53,61.57,42.89,14.48,14.455d10.15(brs,1H,NH),7.95—7.40(m,5H,Ph),6.02—5.91(m,1H,CH),5.25—5.14(m,2H,CH2),4.73—4.70(m,2H,CH2N),4.16(q,J=7.08Hz,2H,OCH2),1.24(t,J=7.05Hz,3H,CH3)158.85,153.75,147.66,132.42,130.98,129.27,128.84,125.56,118.34,61.57,50.40,14.485e10.18(brs,1H,NH),7.96—7.63(m,4H,NO2Ph),4.17(q,J=7.11Hz,2H,OCH2),4.04(q,J=7.23Hz,2H,CH2N),1.35(t,J=7.17Hz,3H,OCH2CH3),1.24(t,J=7.08Hz,3H,CH3)155.15,153.81,148.88,147.28,132.27,130.49,129.86,123.73,123.63,61.67,43.15,14.45,14.365f10.17(brs,1H,NH),7.95—7.63(m,4H,NO2Ph),4.15(q,J=7.08Hz,2H,OCH2),3.98(t,J=7.08Hz,2H,CH2N),1.79—1.75(m,2H,CH2),1.24(t,J=7.08Hz,3H,OCH2CH3),0.84(t,J=7.35Hz,3H,CH3)155.14,153.74,148.92,147.72,132.27,130.47,129.90,123.73,123.71,61.44,49.42,22.16,14.44,10.845g10.19(brs,1H,NH),7.95—7.63(m,4H,NO2Ph),4.15(q,J=7.11Hz,2H,OCH2),4.01(t,J=7.05Hz,2H,CH2N),1.75—1.71(m,2H,CH2),1.29—1.25(m,2H,CH2),1.23(t,J=7.29Hz,3H,OCH2CH3),0.88(t,J=7.26Hz,3H,CH3)155.11,153.74,148.91,147.63,132.25,130.45,129.88,123.71,123.69,61.64,47.56,30.70,19.13,14.44,13.465h10.08(brs,1H,NH),7.77—7.47(m,4H,NO2Ph),5.84—5.72(m,1H,CH),5.08—4.93(m,2H,CH2),4.56—4.54(m,2H,CH2N),3.98(q,J=7.11Hz,2H,OCH2),1.06(t,J=7.11Hz,3H,CH3)157.62,155.49,150.65,149.14,133.24,132.67,131.81,131.44,125.88,124.96,118.91,63.38,51.94,14.705i10.13(brs,1H,NH),7.82—7.42(m,4H,ClPh),4.16(q,J=7.08Hz,2H,OCH2),4.07(q,J=7.20Hz,2H,CH2N),1.38(t,J=7.20Hz,3H,OCH2CH3),1.25(t,J=7.08Hz,3H,CH3)157.37,153.92,146.61,131.50,131.14,130.69,130.51,130.03,127.27,61.57,42.96,14.48,14.425j8.57(brs,1H,NH),7.84—7.26(m,4H,ClPh),4.19(q,J=7.12Hz,2H,OCH2),4.11(t,J=7.37Hz,2H,CH2N),1.93—1.89(m,2H,CH2),1.39—1.35(m,2H,CH2),1.24(t,J=7.06Hz,3H,OCH2CH3),0.94(t,J=7.30Hz,3H,CH3)157.36,153.92,147.00,131.56,131.21,130.75,130.60,130.06,127.36,61.62,47.45,30.86,19.27,14.55,13.575k10.21(brs,1H,NH),7.82—7.40(m,4H,ClPh),6.05—5.91(m,1H,CH),5.26—5.14(m,2H,CH2),4.76—4.73(m,2H,CH2N),4.16(q,J=7.08Hz,2H,OCH2),1.24(t,J=7.08Hz,3H,CH3)159.56,155.61,148.60,134.04,132.76,132.48,131.69,131.47,131.24,127.92,119.05,63.37,51.89,14.735l10.03(brs,1H,NH),7.88—6.99(m,4H,OCH3Ph),4.15(q,J=7.11Hz,2H,OCH2),4.01(q,J=7.20Hz,2H,CH2N),3.97(s,3H,OCH3),1.36(t,J=7.20Hz,3H,OCH2CH3),1.24(t,J=7.08Hz,3H,CH3)160.10,158.69,153.96,146.85,126.99,123.78,114.21,61.51,55.31,42.73,14.93,14.855m10.01(brs,1H,NH),7.88—6.98(m,4H,OCH3Ph),4.15(q,J=7.08Hz,2H,OCH2),3.99(t,J=7.11Hz,2H,CH2N),3.79(s,3H,OCH3),1.78—1.74(m,2H,CH2),1.28—1.24(m,2H,CH2),1.24(t,J=7.08Hz,3H,OCH2CH3),0.80(t,J=7.29Hz,3H,CH3)162.00,160.56,159.19,154.17,146.81,126.84,122.82,113.30,61.60,54.07,30.53,18.97,13.04,12.145n10.13(brs,1H,NH),7.96—7.50(m,4H,ClPh),4.16(q,J=7.08Hz,2H,OCH2),4.05(q,J=7.20Hz,2H,CH2N),1.37(t,J=7.20Hz,3H,OCH2CH3),1.24(t,J=7.08Hz,3H,CH3)157.84,153.90,147.32,133.85,129.98,128.97,127.27,61.61,42.98,14.48,14.425o10.11(brs,1H,NH),7.96—7.50(m,4H,ClPh),4.16(q,J=7.08Hz,2H,OCH2),3.99(t,J=7.02Hz,2H,CH2N),1.83—1.78(m,2H,CH2),1.24(t,J=7.05Hz,3H,OCH2CH3),0.86(t,J=7.35Hz,3H,CH3)157.78,153.80,147.73,133.83,129.96,128.95,127.27,61.56,49.28,22.20,14.46,10.955p8.27(brs,1H,NH),7.99—7.35(m,4H,ClPh),6.06—5.96(m,1H,CH),5.32—5.25(m,2H,CH2),4.80—4.77(m,2H,CH2N),4.26(q,J=7.14Hz,2H,OCH2),1.31(t,J=7.11Hz,3H,CH3)158.51,153.73,147.44,135.22,131.33,129.05,128.68,127.52,119.08,62.60,51.94,14.365q10.26(brs,1H,NH),8.35—8.31(m,4H,NO2Ph),4.18(q,J=7.08Hz,2H,OCH2),4.10(q,J=7.20Hz,2H,CH2N),1.40(t,J=7.20Hz,3H,OCH2CH3),1.25(t,J=7.08Hz,3H,CH3)157.03,153.78,147.86,147.76,136.99,126.53,124.33,61.69,43.23,14.49,14.38

Continued.

Compd.1HNMR(DMSO-d6,300MHz),δ13CNMR(DMSO-d6,75MHz),δ5r10.23(brs,1H,NH),8.34—8.16(m,4H,NO2Ph),4.17(q,J=7.08Hz,2H,OCH2),4.03(t,J=7.02Hz,2H,CH2N),1.84—1.80(m,2H,CH2),1.25(t,J=7.08Hz,3H,OCH2CH3),0.87(t,J=7.35Hz,3H,CH3)156.99,153.70,148.28,147.78,136.98,126.55,124.35,61.66,49.50,22.19,14.47,10.955s10.24(brs,1H,NH),8.35—8.17(m,4H,NO2Ph),4.18(q,J=7.05Hz,2H,OCH2),4.08(t,J=7.11Hz,2H,CH2N),1.82—1.78(m,2H,CH2),1.32—1.28(m,2H,CH2),1.26(t,J=7.11Hz,3H,OCH2CH3),0.90(t,J=7.29Hz,3H,CH3)157.03,153.76,148.23,147.79,137.00,126.58,124.39,61.72,47.71,30.83,19.30,14.52,13.565t10.10(brs,1H,NH),7.99—7.25(m,4H,FPh),4.16(q,J=7.08Hz,2H,OCH2),4.04(q,J=7.20Hz,2H,CH2N),1.37(t,J=7.20Hz,3H,OCH2CH3),1.25(t,J=7.08Hz,3H,CH3)164.43,161.17,157.99,153.93,147.20,127.77,127.66,115.99,115.70,61.60,42.90,14.51,14.475u10.09(brs,1H,NH),7.99—7.25(m,4H,FPh),4.16(q,J=7.02Hz,2H,OCH2),3.98(t,J=7.08Hz,2H,CH2N),1.83—1.79(m,2H,CH2),1.24(t,J=7.05Hz,3H,OCH2CH3),0.86(t,J=7.32Hz,3H,CH3)164.45,161.19,157.97,153.88,147.65,127.80,127.69,116.01,115.72,61.60,49.24,22.27,14.52,11.015v10.17(brs,1H,NH),7.99—7.26(m,4H,FPh),6.01—5.93(m,1H,CH),5.25—5.15(m,2H,CH2),4.72—4.70(m,2H,CH2N),4.16(q,J=7.11Hz,2H,OCH2),1.24(t,J=7.11Hz,3H,CH3)164.46,161.20,158.10,153.72,147.75,132.39,127.81,127.53,118.38,115.97,115.69,61.59,50.41,14.475w10.09(brs,1H,NH),7.99—7.25(m,4H,FPh),4.16(q,J=7.11Hz,2H,OCH2),4.01(t,J=7.11Hz,2H,CH2N),1.80—1.75(m,2H,CH2),1.34—1.26(m,2H,CH2),1.24(t,J=7.08Hz,3H,OCH2CH3),0.88(t,J=7.29Hz,3H,CH3)164.40,161.15,157.93,153.85,147.53,127.76,127.65,115.95,115.66,61.54,47.36,30.80,19.25,14.48,13.505x10.31(brs,1H,NH),8.34—8.16(m,4H,NO2Ph),6.03—5.94(m,1H,CH),5.28—5.17(m,2H,CH2),4.77(d,J=5.73Hz,2H,CH2N),4.17(q,J=7.11Hz,2H,OCH2),1.25(t,J=7.08Hz,3H,CH3)157.13,153.57,148.37,147.82,136.82,132.15,126.59,124.35,118.69,61.69,50.67,14.466a7.62—7.50(m,5H,Ph),4.21(q,J=7.21Hz,2H,OCH2),3.98(q,J=7.08Hz,2H,OCH2),3.95(t,J=7.07Hz,2H,CH2N),1.75—1.71(m,2H,CH2),1.35—1.31(m,2H,CH2),1.18(t,J=7.09Hz,3H,OCH2CH3),1.12(t,J=7.12Hz,3H,OCH2CH3),0.91(t,J=7.30Hz,3H,CH3)160.00,149.77,147.33,145.50,131.14,129.10,128.68,125.27,65.45,60.51,46.32,29.61,19.24,14.65,13.54,13.346b7.69—7.49(m,5H,ClPh),4.13(q,J=7.11Hz,2H,OCH2),3.98(q,J=7.24Hz,2H,OCH2),3.97(t,J=7.10Hz,2H,CH2N),1.31(t,J=7.20Hz,3H,OCH2CH3),1.17(t,J=7.09Hz,3H,OCH2CH3),1.02(t,J=7.13Hz,3H,CH3)159.78,148.90,146.95,143.01,133.47,133.07,132.58,129.51,127.74,125.68,65.31,60.57,42.02,14.66,13.27,13.166c7.51—6.96(m,4H,OCH3Ph),4.26(q,J=7.20Hz,2H,OCH2),4.14(q,J=7.13Hz,2H,OCH2),4.08(t,J=7.23Hz,2H,CH2N),3.78(s,3H,OCH3),1.42(t,J=7.22Hz,3H,OCH2CH3),1.32(t,J=7.17Hz,3H,OCH2CH3),1.20(t,J=7.16Hz,3H,CH3)159.90,157.04,149.13,147.45,143.77,133.11,130.83,121.07,114.74,111.35,65.03,60.47,55.66,41.87,14.71,13.48,13.246d7.51—6.90(m,4H,OCH3Ph),6.03—5.93(m,1H,CH),5.38—5.27(m,2H,CH2),4.65—4.62(m,2H,CH2N),4.26(q,J=7.20Hz,2H,OCH2),4.14(t,J=7.12Hz,2H,OCH2),3.77(s,3H,OCH3),1.31(t,J=7.11Hz,3H,OCH2CH3),1.20(t,J=7.17Hz,3H,CH3)160.62,157.20,149.98,147.57,144.52,132.54,131.02,130.61,121.00,119.48,115.37,110.34,64.77,61.23,55.18,49.32,14.59,13.456e7.54—7.05(m,4H,OCH3Ph),4.22(q,J=7.14Hz,2H,OCH2),3.97(q,J=7.07Hz,2H,OCH2),3.89(t,J=6.96Hz,2H,CH2N),3.82(s,3H,OCH3),1.77—1.73(m,2H,CH2),1.82(t,J=7.08Hz,3H,OCH2CH3),1.52(t,J=7.15Hz,3H,OCH2CH3),0.90(t,J=7.36Hz,3H,CH3)161.40,160.08,149.89,147.48,145.48,130.78,117.18,114.12,65.40,60.45,55.53,48.14,21.05,14.63,13.40,10.966f7.55—7.04(m,4H,OCH3Ph),5.99—5.88(m,1H,CH),5.28—5.21(m,2H,CH2),4.57(d,J=5.55Hz,2H,CH2N),4.22(q,J=7.14Hz,2H,OCH2),3.98(t,J=7.08Hz,2H,OCH2),3.82(s,3H,OCH3),1.20(t,J=7.08Hz,3H,OCH2CH3),1.15(t,J=7.11Hz,3H,CH3)161.42,159.99,149.70,147.43,145.72,131.52,130.83,118.69,117.12,114.13,65.45,60.52,55.56,48.81,14.63,13.42

Continued.

Compd.1HNMR(DMSO-d6,300MHz),δ13CNMR(DMSO-d6,75MHz),δ6g7.61—7.26(m,4H,ClPh),4.32(q,J=7.17Hz,2H,OCH2),4.16(q,J=7.14Hz,2H,OCH2),4.02(t,J=7.41Hz,2H,CH2N),1.85—1.80(m,2H,CH2),1.39—1.33(m,2H,CH2),1.32(t,J=7.11Hz,3H,OCH2CH3),1.26(t,J=7.14Hz,3H,OCH2CH3),0.95(t,J=7.29Hz,3H,CH3)161.01,150.53,147.53,145.13,137.39,130.67,128.68,123.69,65.55,61.35,47.02,30.01,19.70,14.56,13.54,13.48

1.3抑菌活性測試

1.3.1材料測試對象為化合物5a～5x和6a～6g, 選擇多菌靈原藥作為對照. 供試菌種為番茄晚疫病菌(Phytophthora infestans)、 油菜菌核病菌(Sclerotinia scleotiorum)、 蘋果斑點落葉病病原(Ahernaria mali)、 辣椒疫霉病菌(Phytophora capsici)、 西瓜炭疽病菌(Colletetrichum orbiculare)和辣椒灰霉病菌(Botrytis cinerea), 均由中國農業(yè)科學院植物保護研究所提供; 實驗用培養(yǎng)基均為馬鈴薯瓊脂培養(yǎng)基(PDA).

1.3.2實驗方法將目標化合物用二甲亞砜溶解配制成20000mg/L的高濃度母液. 按照一定的濃度梯度, 將測試藥劑加入經滅菌并冷卻至約55 ℃的培養(yǎng)基(PDA)中混勻, 倒入培養(yǎng)皿中, 制成含藥濃度為50mg/L的平板. 實驗設不含藥劑處理的空白對照, 各處理重復3次.以百菌清及多菌靈作為對照藥劑, 采用菌落直徑法進行抑菌活性實驗.

在含藥培養(yǎng)基平板中心分別接一供試病菌的新鮮菌絲塊菌碟, 菌絲朝下. 在25 ℃恒溫培養(yǎng)箱中, 培養(yǎng)至空白對照菌落布滿培養(yǎng)皿2/3以上時,用十字交叉法測量處理的菌落直徑.重復3次, 以其平均數代表菌落的大小.計算藥劑對菌絲生長的抑制率.

藥劑處理對各種病原菌的菌絲生長抑制率公式:

菌絲生長抑制率(%)=(空白菌落直徑-藥劑處理菌落直徑)/(空白菌落直徑-菌碟直徑)×100%

1.4殺蟲活性測試

實驗害蟲為桃蚜[Myzus persicae(Sulzer)], 采自田間的種群; 對照藥劑為吡蟲啉.毒力測定方法如下: 先將原藥用N,N-二甲基甲酰胺配制成母液(質量分數2%), 再用TritonX-100的水溶液(質量分數0.05%)稀釋至測定濃度. 采用藥液浸漬法, 將事先挑選好的帶蚜葉片浸入藥液中, 輕搖10s后取出, 用吸水紙吸去多余藥液, 計數后移入放有濕潤吸水紙保濕的培養(yǎng)皿內, 加保鮮膜加蓋置(25±1) ℃下恢復. 用不含藥劑溶液作空白對照. 48h后在臺式放大鏡下檢查結果. 以毛筆尖輕觸蟲體, 無反應和不能正常爬行者為死亡. 所測藥劑濃度梯度設定為5, 10, 20, 80, 200和400mg/L, 對照藥劑吡蟲啉的濃度梯度設定為40, 20, 10, 2, 1, 0.5和0.2mg/L, 每個梯度平行測定3次. 實驗結果用Probit軟件進行統計分析[20], 計算半致死濃度(LC50).

2結果與討論

2.1目標化合物的合成

以氯甲酸乙酯為原料合成氨基甲酸酯多采用弱堿三乙胺、 吡啶、 碳酸鉀和碳酸鈉等作為縛酸劑, 但在合成化合物5時, 室溫下用弱堿作縛酸劑反應并不能進行. 改用NaOH作縛酸劑后, 以丙酮與水為溶劑, 在室溫下反應能較好地進行, 但產率通常約為50%, 有近1/2的原料未反應, 延長反應時間并不能提高反應收率.由于本實驗以得到產物為目標, 并且未產生副產物, 故未優(yōu)化反應條件.

但對于某些底物, 在相同條件下得到的主要產物為化合物6, 在此情況下化合物5的含量非常少. 進一步研究發(fā)現, 化合物6在酸性條件下容易脫掉4-位的乙氧羰基回到化合物5, 即使在很弱的酸性條件下也容易發(fā)生分解. 考慮到化合物6具有與苯菌靈類似的結構, 故合成了部分化合物6, 并將其活性與化合物5進行了比較.

2.2殺蟲活性

采用浸漬法測定了目標化合物在500μg/mL下對桃蚜的校正致死率, 結果列于表5.可見, 在此濃度下, 大部分目標化合物表現出良好的殺蟲活性. 基于此, 選擇致死率>90%的化合物進一步測定了其精密毒力, 結果列于表6. 分析發(fā)現, 當R2基團為2-氯吡啶-5-甲基時(化合物5a和5b), 活性高于R2為脂肪基團取代的化合物, 其中化合物5b對桃蚜的LC50值為9.49μg/mL, 遠高于其它化合物; 當R1同為芳基時,R2為烯丙基時(化合物5u和5x)的殺蟲活性高于R2為乙基、 正丙基及正丁基取代的化合物(5f, 5j, 5w和5q～5u); 當R1同為芳基取代時, 雙氨基甲酸酯類化合物6的殺蟲活性總體高于單氨基甲酸酯類化合物5, 但化合物5x例外.

Table 5　Corrected mortality of synthesized compounds to Myzus perscicae Sulzer at 500 μg/mL

Table 6　Precision insecticidal activity of selected compounds against Myzus perscicae Sulzer(48 h, LC50)

2.3抑菌活性

采用菌絲生長速率法測定了50μg/mL濃度下目標化合物5和6的離體抑菌活性, 結果列于表7. 可見, 對于化合物5i～5x和5a～5h, 當R1為芳環(huán)時, 苯環(huán)上有吸電子取代時抑菌活性高于苯環(huán)上有給電子基團或無取代基的化合物;R1基團苯環(huán)上為4-Cl取代時(化合物5n～5p), 對多數供試菌種的抑菌活性高于R1基團苯環(huán)上有2-Cl,F及NO2取代的化合物5i～5k, 5q～5x, 但當R1基團苯環(huán)上為2-Cl取代時(化合物5i～5k), 對西瓜炭疽病菌的抑制率明顯高于其它目標化合物; 化合物5r對蘋果斑點落葉病菌的抑制率為74.7%, 高于對照藥劑多菌靈(49.5%); 與化合物5相比, 雙氨基甲酸酯類化合物6的抑菌活性較差, 但當R1基團苯環(huán)上有4-Cl取代時(化合物6g), 對大多供試菌種的抑菌活性高于其它目標化合物.

Table 7　Fungicidal activity of target compounds at 50 μg/mL in vitro inhibitory rate(%)

Continued.

Compd.P.infestansS.scleotiorumA.maliP.capsiciC.orbiculareB.cinereaClgPa5k2.98.635.05.841.62.54.555lNANA3.9NA2.1NA3.515m20.90.421.7NA13.5NA4.425n28.130.325.59.814.10.34.205o36.438.531.5NA16.226.64.685p33.023.233.5NA16.218.84.555q19.711.123.87.212.59.7/5r20.227.674.72.111.214.3/5s22.421.322.35.37.213.4/5t8.212.020.24.912.113.03.805u6.720.424.7NA5.89.24.285v9.415.920.55.97.510.84.155w24.024.029.52.5NA7.94.705x23.119.522.91.7NA1.2/6a19.224.129.4NA9.9NA4.656b14.39.16.54.82.1NA4.306c8.65.513.11.9NANA3.626d6.63.111.9NANANA3.976e7.413.311.0NA4.8NA4.106f7.76.62.7NA1.8NA3.976g46.452.046.018.824.936.65.20Carbendazol10010049.533.874.176.11.29Benomyl2.38

a:lgPmeansoctanol-waterpartitioncoefficient,whichisequaltothelogarithmoftheratioofconcentrationsofanunionizedcompoundbetweenoctanolandwater. ClgPisacalculatedlgPvalueinsilicomethod. “/”meansthattheClgPofcompoundcannotbecalculatedinthesamesilicomethod.NA:noactivity. ---:notested.

總之, 該類化合物的抑菌活性不佳, 盡管具有與多菌靈(Carbendazim)和苯菌靈(Benomyl)類似的結構. 但比較ClgP值可知, 目標化合物的脂溶性遠高于多菌靈和苯菌靈(表7), 甚至高于其未取代的母體化合物4, 其水溶性差可能是造成抑菌活性差的主要原因, 這也為進一步提高其活性提供了新思路.

3結論

采用活性基團拼接的方法, 在1H-1,2,4-三唑-5-胺上引入氨基甲酸酯單元, 合成了一系列新的雜芳基氨基甲酸酯類化合物, 其對桃蚜表現出較高的殺蟲活性, 其中化合物5b的LC50值為9.49μg/mL, 稍遜于對照藥吡蟲啉, 但仍具有進一步研究開發(fā)或作為先導化合物研究的價值. 該類化合物的抑菌活性不強, 僅有個別化合物對某種病原菌表現出較好的活性. 雖然所合成化合物的總體生物活性不夠理想, 但是對此類結構新穎的多取代1H-1,2,4-三唑5-氨基甲酸酯類化合物的活性研究為今后開發(fā)高活性的三唑類化合物提供了新思路.

參考文獻

[1]VelikordovA.V.,IonovaV.A.,DegtyarevO.V.,SukhenkoL.T., Pharm. Chem. J., 2013, 46(12), 715—719

[2]MaH.J.,XieR.L.,ZhaoQ.F.,MeiX.D.,NingJ., J. Agric. Food Chem., 2010, 58(24), 12817—12821

[3]YangJ.C.,MaS.C.,JiangM.F.,ZhangX.,LiuC.L., Agrochemicals Research and Application, 2009, 13(6), 1—5(楊吉春, 馬士存, 姜美峰, 張茜, 劉長令. 農藥研究與應用, 2009, 13(6), 1—5)

[4]HouC.Q.,LiZ.L.,LiuC.L., Pesticides, 2002, 41(6), 41—44(候春青, 李志念, 劉長令.農藥, 2002, 41(6), 41—44)

[5]FanZ.J.,YangZ.K.,ZhangH.K.,MiN.,WangH.,CaiF.,ZuoX.,ZhengQ.X.,SongH.B., J. Agric. Food Chem., 2010, 58(5), 2630—2636

[6]WangZ.J.,SunX.H.,LiuY.F.,ChenB.,ShenS.Q.,JinR.Y.,MaH.X., Chem. J. Chinese Universities, 2015, 36(7), 1315—1320(王子劍, 孫曉紅, 劉源發(fā), 陳邦, 沈生強, 靳如意, 馬海霞. 高等學?；瘜W學報, 2015, 36(7), 1315—1320)

[7]CaoX.F.,HuM.,ZhangJ.,LiF.,YangY.H.,LiuD.L.,LiuS.H., J. Agric. Food Chem., 2009, 57(15), 6914—6919

[8]WangB.L.,ShiY.X.,MaY.,LiuX.H.,LiY.H.,SongH.B.,LiB.J.,LiZ.M., J. Agric. Food Chem., 2010, 58(9), 5515—5522

[9]CaoX.F.,LiF.,HuM.,LuW.C.,YuG.,LiuS.H., J. Agric. Food Chem., 2008, 56(23), 11367—11375

[10]DuanZ.F.,FuL.,LiC.B., Chem. J. Chinese Universities, 2015, 36(1), 102—109(段志芳, 付莉, 李充璧. 高等學?；瘜W學報, 2015, 36(1), 102—109)

[11]LiY.D.,MaoW.T.,FanZ.J.,LiJ.J.,FangZ.,JiX.T.,ZongG.G.,LiF.Y., Chem. Res. Chinese Universities, 2014, 30(3), 390—395

[12]XuL.Z.,BiW.Z.,ShangY.Q.,ZhaiZ.W.,YiX., Chem. Res. Chinese Universities, 2008, 24(3), 299—302

[13]BayrakH.,DemirbasA.,BektasH., Turk. J. Chem., 2010, 34, 835—846

[14]QianY.M.,HamiltonM.,Sidduri.A.,GabrielS.,RenY.L.,PengR.Q.,RamaK.,NarayannanA.,TruittT.,HamidR.,ChenY., J. Med. Chem., 2012, 55, 7920—7939

[15]ColombanoG.,AlbaniC.,OttonelloG.,OttonelloG.,RiberioA., ChemMedChem, 2015, 10, 380—395

[16]RadwanA.M.,El-ZemityS.R., Pest. Manag. Sci., 2001, 57, 707—712

[17]LiuW.D.,DengY.,LanZ.L.,WangX.G.,GaoB.D., Chin. J. Pestic. Sci., 2004, 6(4), 13—17(劉衛(wèi)東, 鄧義, 蘭支利, 王曉光. 高必達. 農藥學學報, 2004, 6(4), 13—17)

[18]JianF.F.,XiaoH.L.,ZhuC.Y.,XuL.Z., J. Heterocyclic. Chem., 2006, 43, 925—929

[19]JiaC.Q.,SuW.C.,XuY.J.,LiuJ.P.,QinZ.H., Chin. J. Org. Chem.,doi: 10.6023/cioc2015/0021(賈長青, 蘇旺蒼, 徐彥軍, 劉吉平, 覃兆海. 有機化學,doi: 10.6023/cioc2015/0021)

[20]RaymondM., Cah. Orstom Ser. Ent. Med. Parasitol., 1985, 23, 117—121

(Ed.:P,H,Y,K)

?SupportedbytheNationalNaturalScienceFoundationofChina(No.31272076).

doi:10.7503/cjcu20160065

收稿日期:2016-01-26. 網絡出版日期: 2016-04-18.

基金項目:國家自然科學基金(批準號: 31272076)資助.

中圖分類號O626

文獻標志碼A

Synthesis,StructuralCharacterization,InsecticidalandFungicidalActivityof(1H-1,2,4-Triazol-5-yl)carbamates?

JIAChangqing1,YANGDongyan1,CHEChuanliang1,MAYongqiang1,RUIChanghui2,YANXiaojing2,QINZhaohai1*

(1. College of Science, China Agricultural University, Beijing 100193, China;2. Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)

AbstractA series of novel (1H-1,2,4-triazol-5-yl)carbamates(5 and 6) was synthesized via the method of combining the crucial sub-structures of carbamates and 1H-1,2,4-triazole-5-amines in one molecules. Their structures were characterized by nuclear magnetic resonance(NMR), infrared(IR) spectrometry, high resolution mass spectrometry(HRMS) and elemental analysis. The preliminary insecticidal activity assays indicated that the lethal rates of target compounds to Myzus persicae were good at the concentration of 500 μg/mL. And the lethal rate of several compounds was more than 90%. Further research of these compounds against Myzus persicae indicated that compound 5b showed considerably high activity with a LC50of 9.49 μg/mL, which confirmed that these compounds are valuable for further study. The preliminary fungicidal activity assays indicated that these compounds exhibited low fungicidal activity against tested fugi in vitro. At a dose of 50 μg/mL, only a few of compounds gave limited inhibition on mycelium growth. In addition, the structure-activity relationship was also discussed.

Keywords1H-1,2,4-triazole-5-amine; Carbamate; Insecticidal activity; Fungicidal activity

聯系人簡介: 覃兆海, 男, 博士, 教授, 博士生導師, 主要從事新農藥創(chuàng)制研究.E-mail:qinzhaohai@263.net