CN116210706A - 生物碱polyaurine B衍生物在抗植物病毒和病菌中的应用 - Google Patents
生物碱polyaurine B衍生物在抗植物病毒和病菌中的应用 Download PDFInfo
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- CN116210706A CN116210706A CN202310053505.3A CN202310053505A CN116210706A CN 116210706 A CN116210706 A CN 116210706A CN 202310053505 A CN202310053505 A CN 202310053505A CN 116210706 A CN116210706 A CN 116210706A
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Abstract
本发明生物碱polyaurine B衍生物在抗植物病毒和病菌中的应用,涉及含有3‑氨基‑5‑苯基‑1,2,4‑噻二唑的生物碱polyaurine B衍生物的杀生剂,具体地说是二十八种生物碱polyaurine B衍生物在抗植物病毒和病菌用作抗植物病毒剂和用作杀植物病菌剂,所述用作抗植物病毒剂是用于烟草花叶病毒;所述用作杀植物病菌剂是用于抑制油菜菌核病菌、花生褐斑病菌、黄瓜枯萎病菌、水稻纹枯病菌、番茄早疫病菌、苹果轮纹病菌、小麦纹枯病菌或水稻稻瘟病菌等8种植物病菌。
Description
技术领域
本发明的技术方案涉及含有3-氨基-5-苯基-1,2,4-噻二唑的生物碱polyaurineB衍生物的的杀生剂,具体地说是生物碱polyaurine B衍生物在抗植物病毒和病菌中的应用。
背景技术
Polyaurine类生物碱是一类具有新颖结构的噻二唑生物碱,通过胺基官能团在噻二唑C-5位连接一个酰胺结构。从自然界提取的或者人工合成的含有该类核心骨架的化合物有polyaurine B、dendrodoine、P2Y2 Receptor Inhibitor、Dendrodoine cytotoxic、SCH-202676及Neuropeptide Y Receptor Antagonit等polyaurines生物碱(Org.Lett.,2013,15,9,2230-2233;Org.Lett.,2009,11,24,5666-5669;Mar.Drugs.,2019,19,278;J.Nat.Prod.2020,83,1721-1724),如结构式一所示。
自2019年Marcello等人第一次报道从海洋被膜多糖的代谢成分中分离得到polyaurine类生物碱以来(Mar.Drugs.,2019,19,278),后续证实polyaurine类生物碱对哺乳动物的细胞生长以及寄生虫的活力有显著的影响,还有杀菌、抗癌、抗病毒等效用(RussChem Bull,2021,3,27,3114-3116)。
通过文献调研,由于polyaurine类生物碱自然界含量低,且合成收率低、成本高,研究人员对其应用研究主要集中在医药特别是抗肿瘤方面,在控制病虫害方面还鲜有报道。随着polyaurine类生物碱合成新方法的不断探索,Daniel G等人(J.Nat.Prod.,2020,83,1721-1724;见反应式一)报道了合成pityriacitrin及其衍生物制备的新方法,极大的降低了合成的难度及成本。因此,通过对其结构修饰和改造,将其应用于控制农业生产中的病虫害,不仅可以拓宽polyaurine类衍生物的生物活性谱,而且对农作物的保护具有非常重要的应用价值。
发明内容
本发明的目的是提供生物碱polyaurine B衍生物在抗植物病毒和病菌中的应用,首次发现生物碱polyaurine B衍生物具有很好的抗植物病毒和病菌活性,扩大了生物碱polyaurine B衍生物作为生物农药的应用范围。
本发明解决该技术问题所采用的技术方案是:
生物碱polyaurine B衍生物在抗植物病毒和病菌中的应用,具体地说是以下1~28种化学结构式所示的生物碱polyaurine B衍生物衍生物用作抗植物病毒剂和用作杀植物病菌剂,
上述生物碱polyaurine B衍生物的应用,其特征在于:用作抗植物病毒剂,所述植物病毒为烟草花叶病毒。
上述生物碱polyaurine B衍生物的应用,其特征在于:用作杀植物病原菌剂,所述植物病原菌为油菜菌核病菌、花生褐斑病菌、黄瓜枯萎病菌、水稻纹枯病菌、番茄早疫病菌、苹果轮纹病菌、小麦纹枯病菌或水稻稻瘟病菌等8种植物病菌。
与现有技术相比,本发明具有的突出的实质性特点和显著进步如下:
(1)本发明针对polyaurine B制备过程中需要用到剧毒化学品氯甲酸甲酯这一问题,在保证其生物活性的同时通过原料的替换,以廉价易得的酰氯为原料,经酰胺化得到生物碱polyaurine类衍生物,避免使用了高毒性试剂;
(2)首次发现生物碱polyaurine类衍生物具有很好的抗植物病毒活性,在同等测试条件下,多数优于商品化品种病毒唑,化合物19和23在100μg/mL时表现出比宁南霉素更高的抗TMV活性;此外,polyaurine B衍生物对8种常见农业病原体均具有广谱抑制活性。化合物5、11、12、27表现出中等到良好的杀菌活性,对一些真菌的活性超过90%,扩大了生物碱polyaurine B衍生物作为生物农药的应用范围。
本发明首次发现生物碱polyaurine B衍生物具有很好的抗植物病毒活性和广谱的杀植物病原菌活性,得到的polyaurine B衍生物表现出优良的生物活性,部分化合物抗病毒活性和杀菌活性取得了意想不到的效果和突破性进展。
具体实施方式
实施例1
化学结构式1~28所示生物碱polyaurine B衍生物的制备方法如反应式二所示:
具体操作步骤如下:
第一步,将4-甲氧基苯甲酸甲酯(498.5mg,3.0mmol)、P4S10(513mg,0.7mmol)和二聚硅氧烷(5mL)回流于对二甲苯(10mL)中。根据TLC监测,连续搅拌反应物直至完成。反应完全后,溶剂用旋转蒸发器蒸干。残渣中多次加入甲醇提取产物。真空脱去溶剂,经柱层析提纯(洗脱剂为石油醚:乙酸乙酯=60:1),得到492.0mg棕色固体产物,收率为89%;该红棕色固体物质的相关参数为:1H NMR(400MHz,DMSO-d6)δ8.12(d,J=8.9Hz,2H),7.01(d,J=8.9Hz,2H),4.23(s,3H),3.84(s,3H);13C NMR(101MHz,DMSO-d6)δ211.2,164.1,131.3,131.0,114.3,59.8,56.2.确定该中间产物为4-甲氧基苯硫代苯甲酸甲酯。
第二步,氢化钠(60%矿物油,140mg,3.5mmol)在0℃下加入到甲基胍盐酸盐(264mg,2.4mmol)的THF-DMF(2:1,12mL)溶液中,溶液搅拌10分钟后,将4-甲氧基苯硫代苯甲酸甲酯(400mg,2.2mmol)的THF溶液(4mL)逐滴滴加,混合物50℃搅拌18h。反应完成后,混合物冷却到0℃,用水(20mL)淬灭,水层用乙酸乙酯(3×10mL)萃取。收集有机层,用无水Na2SO4干燥,过滤,真空浓缩,经柱层析提纯(洗脱剂为石油醚:乙酸乙酯=1:1),得到294.7mg淡黄色固体产物,收率为60%;该淡黄色固体物质的相关参数为:1HNMR(400MHz,DMSO-d6)δ9.11(s,1H),8.21(d,J=8.6Hz,2H),6.86(d,J=8.7Hz,2H),3.78(s,3H),2.83(s,3H);13C NMR(101MHz,DMSO-d6)δ198.0,164.3,161.5,130.3,112.8,55.7,28.2;确定该中间产物为4-甲氧基-N-(N-甲基氨基甲酰胺基)苯并硫酰胺。
第三步,将碳酸铯(594mg,1.8mmol)和4-甲氧基-N-(N-甲基氨基甲酰胺基)苯并硫酰胺(215mg,1.0mmol)溶解在DMF(5mL)中添加到溶有Cu(OAc)2·H2O(30mg,0.2mmol)的DMF(3mL)溶液中,反应混合物在开放的烧瓶中80℃下搅拌3h,冷却至室温,真空浓缩。残渣用乙酸乙酯(10mL)溶解,用水(2×10mL)洗涤,收集有机层用无水Na2SO4干燥,过滤,真空浓缩,经柱层析提纯(洗脱剂为石油醚:乙酸乙酯=3:1),得到238.9mg白色固体化合物1,收率为60%;该白色固体物质的相关参数为:1H NMR(400MHz,DMSO-d6)δ7.82(d,J=8.7Hz,2H),7.33(s,1H),7.08(d,J=8.7Hz,2H),3.84(s,3H),2.86(d,J=4.7Hz,3H);13C NMR(101MHz,DMSO-d6)δ185.6,171.8,162.6,128.9,123.5,115.3,56.0,30.0;HRMS(ESI):calcd forC10H11N3OS:m/z[M+H]+,222.0692;found:222.0696。
第四步,将碘甲烷、溴代烷烃或氯甲酸酯或(1.5mL)在0℃下滴加到5-(4-甲氧基苯基)-N-甲基-1,2,4-噻二唑-3-胺(221.0mg,1.0mmol)中,混合物回流搅拌12h,经柱层析提纯(洗脱剂为石油醚:乙酸乙酯=20:1),得到化合物2~28。
化合物2,白色固体,收率为83%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.84(d,J=8.7Hz,2H),6.95(d,J=8.6Hz,2H),3.87(s,3H),3.24(s,6H);13C NMR(100MHz,CDCl3)δ186.2,172.3,162.3,128.7,124.1,114.4,55.5,39.1;HRMS(ESI):calcd forC11H13N3OS:m/z[M+H]+,236.0858;found(ESI+):236.0852。
化合物3,黄色油状物,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.84(d,J=8.5Hz,2H),6.95(d,J=8.5Hz,2H),3.87(s,3H),3.66(t,J=7.3Hz,2H),3.21(s,3H),1.64(q,J=7.4Hz,2H),1.38(q,J=7.4Hz,2H),0.96(t,J=7.3Hz,3H);13C NMR(100MHz,CDCl3)δ185.7,171.5,162.2,128.7,124.0,114.3,55.5,51.4,36.7,29.7,20.0,14.0;HRMS(ESI):calcd for C14H19N3OS:m/z[M+H]+,278.1326;found(ESI+):277.1322。
化合物4,白色固体,收率为90%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.86(d,J=8.2Hz,2H),7.32(s,5H),6.96(d,J=8.3Hz,2H),4.93(s,2H),3.87(s,3H),3.18(s,3H);13C NMR(100MHz,CDCl3)δ186.3,171.8,162.4,138.3,129.1,128.8,128.6,127.7,127.3,124.0,114.4,55.3,36.3;HRMS(ESI):calcd for C17H17N3OS:m/z[M+H]+,312.1160;found(ESI+):312.1165。
化合物5,棕色固体,收率为90%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.88(d,J=8.1Hz,2H),6.97(d,J=8.1Hz,2H),5.14–5.04(m,1H),3.87(s,3H),3.53(s,3H),1.33(d,J=5.6Hz,6H);13C NMR(101MHz,DMSO-d6)δ187.0,166.3,163.1,154.0,129.3,122.9,115.5,70.3,56.1,36.1,22.2;HRMS(ESI):calcd for C14H17N3O3S:m/z[M+H]+,222.0692;found:222.0696。
化合物6,黄色固体,收率为92%;该物质的相关参数为:1H NMR(400MHz,DMSO-d6)δ7.94(d,J=8.5Hz,2H),7.11(d,J=8.4Hz,2H),3.94(d,J=6.4Hz,2H),3.85(s,3H),3.40(s,3H),1.90(dt,J=13.2,6.6Hz,1H),0.91(d,J=6.7Hz,6H);13C NMR(101MHz,CDCl3)δ187.1,166.2,162.7,154.5,128.9,123.3,114.6,72.6,55.6,36.0,27.9,19.1;HRMS(ESI):calcd for C15H19N3O3S:m/z[M+H]+,308.1069;found:308.1064。
化合物7,白色固体,收率为90%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.86(d,J=8.2Hz,2H),7.45(d,J=7.1Hz,1H),7.36(s,2H),7.33(d,J=8.7Hz,2H),6.97(d,J=8.2Hz,2H),5.31(s,2H),3.88(s,3H),3.58(s,3H);13C NMR(101MHz,CDCl3)δ187.2,166.0,162.8,154.3,136.1,128.9,128.5,128.1,128.0,123.3,114.6,68.1,55.6,36.1;HRMS(ESI):calcd for C18H17N3O3S:m/z[M+H]+,322.1223;found:322.1220。
化合物8,白色固体,收率为80%;该白色固体物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.88(d,J=8.7Hz,2H),7.00(d,J=8.7Hz,2H),3.89(s,3H),3.54(s,3H),2.52(s,3H);13C NMR(101MHz,CDCl3)δ187.5,171.8,167.1,162.9,128.9,123.0,114.7,55.6,34.6,25.2;
HRMS(ESI):calcd for C12H13N3O2S:m/z[M+H]+,264.0807;found:264.0801。
化合物9,淡黄色固体,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.88(d,J=8.1Hz,2H),7.00(d,J=8.1Hz,2H),3.89(s,3H),3.53(s,3H),2.86(q,J=6.9Hz,2H),1.20(t,J=7.1Hz,3H);13C NMR(101MHz,CDCl3)δ187.4,175.3,167.1,162.9,128.9,123.1,114.7,55.6,34.7,29.9,9.7;HRMS(ESI):calcd for C13H15N3O2S:m/z[M+H]+,278.0960;found:278.0958。
化合物10,白色固体,收率为83%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ8.00–7.69(m,2H),7.00(d,J=8.6Hz,2H),3.89(s,3H),3.56(d,J=30.1Hz,3H),2.80(t,J=7.4Hz,2H),1.73(dq,J=14.6,7.1Hz,2H),0.95(t,J=7.3Hz,3H);13C NMR(101MHz,CDCl3)δ187.4,174.4,167.1,162.9,128.9,123.1,114.7,55.6,38.3,34.7,18.9,14.0;HRMS(ESI):calcd for C14H17N3O2S:m/z[M+H]+,292.1118;found:292.1114。
化合物11,淡黄色固体,收率为85%;经测定,该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.88(d,J=8.6Hz,2H),7.01(d,J=8.6Hz,2H),3.89(s,3H),3.50(s,3H),1.59(s,1H),1.20(d,J=6.7Hz,6H);13C NMR(101MHz,CDCl3)δ187.5,178.9,167.2,163.0,128.9,123.1,114.7,55.6,35.1,33.1,20.0;HRMS(ESI):calcd for C14H17N3O2S:m/z[M+H]+,292.1116;found:292.1114。
化合物12,白色固体,收率为87%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.88(d,J=8.6Hz,2H),7.00(d,J=8.6Hz,2H),3.89(s,3H),3.52(s,3H),2.73(d,J=6.9Hz,2H),2.17(dt,J=13.1,6.7Hz,1H),0.95(d,J=6.6Hz,6H);13C NMR(101MHz,CDCl3)δ187.4,173.9,167.2,162.9,128.9,123.1,114.7,55.6,45.0,34.7,25.8,22.7;HRMS(ESI):calcd for C15H19N3O2S:m/z[M+H]+,306.1274;found:306.1271。
化合物13,白色固体,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.90(d,J=7.9Hz,2H),7.00(d,J=8.0Hz,2H),3.89(s,3H),3.35(s,3H),1.15(s,9H);13CNMR(101MHz,CDCl3)δ188.0,179.8,168.2,163.1,128.9,122.9,114.7,55.6,41.3,37.8,28.5;HRMS(ESI):calcd for C15H19N3O2S:m/z[M+H]+,306.1276;found:306.1271。
化合物14,黄色固体,收率为78%;该物质的相关参数为:1H NMR(400MHz,Chloroform-d)δ7.63–7.59(m,2H),7.42(d,J=8.0Hz,2H),7.35(d,J=7.2Hz,1H),7.29(d,J=7.5Hz,2H),6.92(d,J=8.6Hz,2H),3.86(s,3H),3.68(s,3H);13C NMR(101MHz,CDCl3)δ187.2,171.6,167.5,162.9,136.7,133.6,130.4,128.8,127.9,122.9,114.6,55.6,35.5;HRMS(ESI):calcd for C17H15N3O2S:m/z[M+H]+,326.0955;found:326.0958。
化合物15,白色固体,收率为88%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.49(d,J=7.5Hz,2H),7.39(s,1H),7.29(s,3H),6.89(s,2H),3.85(s,3H),3.74(s,3H);13CNMR(101MHz,CDCl3)δ168.8,162.8,133.4,132.4,131.5,130.7,130.1,129.2,128.8,128.6,126.6,122.9,114.5,55.5,34.2;HRMS(ESI):calcd for C17H14ClN3O2S:m/z[M+H]+,360.0573;found:360.0568。
化合物16,棕色固体,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.64(d,J=7.7Hz,2H),7.48(s,1H),7.33(d,J=7.3Hz,1H),7.20(d,J=8.3Hz,2H),6.93(d,J=7.8Hz,2H),3.86(s,3H),3.67(s,3H);13C NMR(101MHz,CDCl3)δ187.5,170.1,167.0,163.0,138.5,134.1,130.3,129.2,128.8,128.2,125.8,122.8,114.6,55.6,35.50;HRMS(ESI):calcd for C17H14ClN3O2S:m/z[M+H]+,360.0570;found:360.0568。
化合物17,棕色固体,收率为87%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.48(d,J=8.2Hz,3H),7.35(dd,J=14.3,7.3Hz,2H),7.21(t,J=7.5Hz,1H),6.88(d,J=8.3Hz,2H),3.85(s,3H),3.74(s,3H);13C NMR(101MHz,CDCl3)δ186.9,169.5,165.7,162.8,139.9,132.4,130.2,128.8,128.6,127.1,122.9,119.4,114.5,55.6,34.2;HRMS(ESI):calcd for C17H14BrN3O2S:m/z[M+H]+,404.0067;found:404.0063。
化合物18,白色固体,收率为82%;该物质的相关参数为:1H NMR(400MHz,Chloroform-d)δ7.70(s,3H),7.43(d,J=8.3Hz,2H),7.33(d,J=6.4Hz,1H),6.86(d,J=8.3Hz,2H),3.84(s,3H),3.75(s,3H);13C NMR(101MHz,CDCl3)δ187.1,169.1,165.7,162.8,160.7,132.8,132.1,131.4,131.3,128.8,128.7,127.4,114.5,55.6,34.3。
化合物19,黄色固体,收率为85%;该物质的相关参数为:1H NMR(400MHz,Chloroform-d)δ7.51(d,J=8.1Hz,2H),7.21(t,J=7.3Hz,1H),7.15(d,J=10.9Hz,2H),7.07(t,J=7.1Hz,1H),6.89(d,J=8.2Hz,2H),3.85(s,3H),3.70(s,3H),2.36(s,3H);13CNMR(101MHz,CDCl3)δ186.9,171.9,1667,162.8,137.7,135.4,130.1,129.0,128.8,126.4,125.3,123.0,114.5,55.5,34.5,19.5;HRMS(ESI):calcd for C18H17N3O2S:m/z[M+H]+,340.1116;found:340.1114。
化合物20,红棕色固体,收率为70%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.85(d,J=8.1Hz,2H),7.25(s,2H),7.21(d,J=7.3Hz,3H),6.99(d,J=8.3Hz,2H),4.26(s,2H),3.89(s,3H),3.55(s,3H);13C NMR(101MHz,CDCl3)δ187.6,172.4,166.9,163.0,135.4,129.4,128.9,128.4,126.7,123.0,114.7,55.6,42.9,35.0;HRMS(ESI):calcd forC18H17N3O2S:m/z[M+H]+,340.1119;found:340.1114。
化合物21,棕色固体,收率为83%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.78(d,J=8.4Hz,2H),7.39(d,J=4.8Hz,1H),7.08–7.04(m,1H),6.97(d,J=8.4Hz,2H),6.92–6.87(m,1H),3.87(s,3H),3.61(s,3H);13C NMR(101MHz,CDCl3)δ187.9,167.5,164.3,163.0,138.2,130.9,130.3,128.9,126.8,122.9,114.7,55.6,36.2;HRMS(ESI):calcd forC15H13N3O2S2:m/z[M+H]+,332.0520;found:332.0522。
化合物22,淡黄色油状物,收率为82%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.89(d,J=7.9Hz,2H),7.00(d,J=8.1Hz,2H),3.89(s,3H),3.54(s,3H),2.61–2.50(m,1H),1.15(s,2H),0.88–0.84(m,2H);13C NMR(101MHz,CDCl3)δ187.4,175.1,167.4,162.9,128.9,123.1,114.7,55.6,35.2,14.0,9.7;HRMS(ESI):calcd for C14H15N3O2S:m/z[M+H]+,290.0955;found:290.0958。
化合物23,白色固体物质,收率为80%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.90–7.87(m,2H),7.01–6.98(m,2H),3.89(s,3H),3.53(s,3H),2.55(s,1H),1.31–0.75(m,8H);13C NMR(101MHz,CDCl3)δ187.5,175.1,167.4,162.9,128.9,123.1,114.7,55.6,35.2,14.0,9.7,9.1;HRMS(ESI):calcd for C16H19N3O2S:m/z[M+H]+,236.0858;found:236.0852。
化合物24,白色固体物质,收率为70%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.99–7.91(m,2H),7.17(t,J=8.0Hz,2H),5.10(p,J=6.0Hz,1H),3.54(s,3H),1.34(d,J=6.1Hz,6H);13C NMR(101MHz,CDCl3)δ186.1,166.5,163.7,153.9,129.4,126.9,116.4,70.5,35.9,22.1;HRMS(ESI):calcd for C13H14FN3O2S:m/z[M+H]+,296.0870;found:296.0864。
化合物25,淡黄色固体物质,收率为86%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.87(d,J=8.1Hz,2H),7.46(d,J=8.2Hz,2H),5.17–5.02(m,1H),3.54(s,3H),1.37–1.29(m,6H);13C NMR(101MHz,CDCl3)δ186.1,166.6,153.8,138.2,129.6,128.9,128.4,22.1;HRMS(ESI):calcd for C13H14ClN3O2S:m/z[M+H]+,312.0571;found:312.0568。
化合物26,白色固体物质,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.84–7.78(m,2H),7.63(d,J=8.0Hz,2H),5.10(dt,J=11.8,5.8Hz,1H),3.54(s,3H),1.34(d,J=6.0Hz,6H);13C NMR(101MHz,CDCl3)δ186.2,166.7,153.9,132.6,129.4,128.5,126.6,70.5,35.9,22.1;HRMS(ESI):calcd for C13H14BrN3O2S:m/z[M+H]+,356.0067;found:356.0063。
化合物27,黄色固体物质,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.82(d,J=7.6Hz,2H),7.29(s,2H),5.17–5.04(m,1H),3.54(s,3H),2.42(s,3H),1.33(d,J=5.9Hz,6H);13C NMR(101MHz,CDCl3)δ187.5,166.4,154.0,142.7,129.9,127.9,127.1,70.3,35.9,22.1,21.7;HRMS(ESI):calcd for C14H17N3O2S:m/z[M+H]+,292.1110;found:292.1114。
化合物28,白色固体物质,收率为85%;该物质的相关参数为:1H NMR(400MHz,CDCl3)δ7.48(s,2H),7.39(t,J=7.7Hz,1H),7.07(d,J=7.6Hz,1H),5.14–5.05(m,1H),3.89(s,3H),3.55(s,3H),1.37–1.32(m,6H);13C NMR(101MHz,CDCl3)δ187.3,166.5,160.2,154.0,131.7,130.4,119.7,118.3,111.8,70.4,55.6,35.9,22.1;HRMS(ESI):calcd forC14H17N3O3S:m/z[M+H]+,308.1068;found:308.1064。
实施例2
上述生物碱polyaurine B衍生物1~28所示个体化合物的抗烟草花叶病毒活性的测定,测定程序如下:
第一步,烟草花叶病毒提纯及浓度测定:
烟草花叶病毒提纯及浓度测定参照南开大学元素所生测室编制烟草花叶病毒SOP规范执行,病毒粗提液经2次聚乙二醇离心处理后,测定浓度为20μg/mL,4℃冷藏备用;
第二步,上述生物碱polyaurine B衍生物1~28所示个体化合物药剂溶液的配制:
分别称量生物碱polyaurine B衍生物1~28所示个体化合物40mg作为原药,然后分别在该各个原药中加入DMF 0.4mL进行溶解,制得1×105μg/mL母液,再用质量百分比浓度为1‰的吐温80水溶液稀释至测试浓度为500μg/mL或100μg/mL,由此配制得上述生物碱polyaurine B衍生物1~28所示个体化合物药剂溶液,另外取宁南霉素制剂直接兑水稀释作为对比物;
第三步,活体保护作用:
分别选十份长势均匀一致的3–5叶期珊西烟,分别全株喷雾施第二步配制得的上述生物碱polyaurine B衍生物1~28所示个体化合物药剂溶液,每处理3次重复,并设置质量百分比浓度为1‰的吐温80水溶液对照,24h后,叶面撒布500目金刚砂,用毛笔蘸取病毒液,在全叶面沿支脉方向轻擦2次,叶片下方用手掌支撑,病毒浓度10μg/mL,接种后用流水冲洗,于常温光照条件下保湿培养,每3片叶为1次重复,重复3次,3天后记录病斑数,计算防效;;
第四步,活体治疗作用:
分别选十份长势均匀一致的3–5叶期珊西烟,分别用毛笔全叶接种病毒,病毒浓度为10μg/mL,接种后用流水冲洗,叶面收干后,全株喷雾施第二步配制的上述生物碱polyaurine B衍生物1~28所示个体化合物药剂溶液,每处理3次重复,并设置质量百分比浓度为1‰的吐温80水溶液对照,3天后记录病斑数,计算防效;
第五步,活体钝化作用:
分别选十份长势均匀一致的3–5叶期珊西烟,分别将第二步配制的上述生物碱polyaurine B衍生物1~28所示个体化合物药剂溶液与等体积的病毒汁液混合钝化30min后,摩擦接种,病毒浓度为20μg/mL,接种后即用流水冲洗,重复3次,设置质量百分比浓度为1‰的吐温80水溶液对照,3天后数病斑数,计算结果;
上述生物碱polyaurine B衍生物1~28所示个体化合物的抗烟草花叶病毒活性的测定结果见表1。
表1.生物碱polyaurine B衍生物1~28所示个体化合物的抗TMV活性测试结果:
表1生物活性测试结果发现,生物碱polyaurines及其衍生物具有很好的抗植物病毒活性。在同等测试条件下,多数优于商品化品种病毒唑,部分化合物在100μg/mL时表现出比宁南霉素更高的抗TMV活性,具备开发价值。
实施例3
上述生物碱polyaurine B衍生物1~28中的个体化合物的抗菌活性测试,离体杀菌测试,测定程序如下:
菌体生长速率测定法即平皿法:分别将3mg上述生物碱polyaurine B衍生物1~28中的个体化合物溶解在0.03mL丙酮内,然后用含有200μg/mL吐温80的水溶液稀释至测试浓度为50mg/kg,然后各吸取1mL药液注入与之对应的培养皿内,再分别加入9mL培养基,摇匀后制成50μg/mL的含药平板,以添加1mL灭菌纯净水的平板做空白对照,用直径4mm的打孔器沿菌丝外缘切取菌盘,移至上述含药平板上,每处理重复三次,将培养皿放在24±1℃恒温培养箱内培养,48h后调查各处理菌盘扩展直径,求平均值,与空白对照比较计算相对抑菌率。
上述生物碱polyaurine B衍生物1~28中的个体化合物的离体杀真菌活性的结果见表2。
表2.生物碱polyaurine B衍生物1~28中的个体化合物的离体杀菌活性测试结果
表2说明生物碱polyaurine B衍生物1~28对8种常见农业病原体均具有广谱抑制活性。部分化合物表现出中等到良好的杀菌活性,特别是化合物1、5、11、12、27对一些病原菌的抑制率超过90%。
上述实施例中的百分比均为质量百分比。
上述实施例中所涉及的原料和试剂均由商购获得,化学反应工艺是本技术领域的技术人员所能掌握的。
本发明未尽事宜为公知技术。
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