芳香丙烯酰胺-三氮唑类化合物及其制备方法及和应用

1.本发明公开了一种新型芳香丙烯酰胺-三氮唑类化合物、其制备方法及其在抗真菌药物制备中的用途，属于药物化学领域。


背景技术：

2.近几十年来，随着肿瘤化疗、器官移植及免疫系统功能低下人群数量的增加，侵袭性真菌感染的发病率及死亡率都呈现明显上升趋势，并导致每年150～200万人死亡。抗生素滥用、真菌抑菌药的广泛使用以及介入性插管手术的广泛开展导致真菌耐药甚至是多药耐药现象日趋严重(fisher,m.c.；hawkins,n.j.；sanglard,d.；gurr,s.j.science 2018,360,739.)，基于抗真菌药物类型的有限性以及各类型均出现耐药现象，研发高效、抗菌谱广、对耐药菌有效的抗真菌新药已迫在眉睫，并具有重要的社会意义和应用价值。
3.真菌与哺乳动物同属真核生物，真菌细胞膜的构成需要麦角固醇，而哺乳动物细胞物构成需要胆固醇，这是二者之间的最显著区别(watt,k.；manzoni,p.；cohen-wolkowiez,m.；rizzollo,s.；boano,e.；jacqz-aigrain,e.；benjamin jr,d.k..current drug metabolism 2013,14,193.)，研究表明麦角固醇在维持真菌细胞膜的流动性、完整性、调解细胞形态和影响真菌侵袭过程方面有着重要作用。由于真菌只能依靠自身合成得到麦角固醇，故抑制其合成麦角固醇所需关键酶—羊毛甾醇14α-脱甲基酶(lanosterol 14α-demethylase，以下简称cyp51)的活性是抑制真菌生长的有效方式(shrestha,s.k.；garzan,a.；garneau-tsodikova,s.,eur j med chem 2017,133,309-318.)。基于唑类结构的cyp51抑制剂是目前使用最广泛、市场份额最大的抗真菌药物，以现有药物与cyp51的共晶模型为依据，在保留咪唑环或三氮唑环结构的基础上合成新类型的cyp51抑制剂是寻求高活性抗真菌分子的主要途径和解决耐药问题的有效办法。(fuentefria,a.m.；pippi,b.；dalla lana,d.f.；donato,k.k.；de andrade,s.f.letters in applied microbiology 2018,66,2；li,z.；liu,n.；tu,j.；ji,c.；han,g.；sheng,c.acs infectious diseases 2019,5,1376；fuentefria,a.m.；pippi,b.；dalla lana,d.f.；donato,k.k.；de andrade,s.f.letters in applied microbiology 2018,66,2；thamban chandrika,n.；shrestha,s.k.；ngo,h.x.；tsodikov,o.v.；howard,k.c.；garneau-tsodikova,s.journal of medicinal chemistry 2018,61,158；benhamou,r.i.；bibi,m.；berman,j.；fridman,m.localizing antifungal drugs to the correct organelle can markedly enhance their efficacy.angewandte chemie international edition 2018,57,6230.)
4.喹啉结构广泛存在于具有药理活性的天然产物分子和上市药物分子当中，在分子结构修饰中能起到提高活性、改善药代动力学的作用(boger,d.l.；tse,w.c.bioorganic&medicinal chemistry,2001,9,2511-2518；wang,m.,gao,m.,miller,k.d.,sledge,g.w.,hutchins,g.d.,and zheng,q.h.eur j med chem,2009,44,2300-2306)。同时，含有喹啉结构的噻唑橙类物质因具有抗肿瘤、免疫、抗炎等多种药理活性而备受药学家的关注。
5.因此，以三氮唑环和喹啉结构类化合物利用拼接技术合成新一类化合物，进行抗
基)丙烯酰胺
27.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(吡啶-3-基)丙烯酰胺
28.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(吡啶-4-基)丙烯酰胺
29.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(萘-1-基)丙烯酰胺
30.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(4-吗啉基苯基)丙烯酰胺
31.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(1h-吲哚-3-基)丙烯酰胺
32.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹喔啉-2-基)丙烯酰胺
33.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹啉-2-基)丙烯酰胺
34.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-甲氧基喹啉-2-基)丙烯酰胺
35.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-甲基喹啉-2-基)丙烯酰胺
36.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-氟喹啉-2-基)丙烯酰胺
37.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-(三氟甲基)喹啉-2-基)丙烯酰胺
38.3-(6-氯喹啉-2-基)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-丙烯酰胺
39.乙基-2-(3-((2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)氨基)-3-氧代丙基-1-烯-1-基)喹啉-6-羧酸酯
40.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-苯基喹啉-2-基)丙烯酰胺
41.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(8-甲氧基喹啉-2-基)丙烯酰胺
42.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-溴喹啉-2-基)丙烯酰胺
43.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-碘喹啉-2-基)丙烯酰胺
44.n-(2-(2,4-二氯苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹啉-2-基)丙烯酰胺
45.n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹啉-3-基)丙烯酰胺
46.47.[0048][0049]
制备所述芳香丙烯酰胺-三氮唑类物质的方法，路线如下所示：
[0050][0051]
r1、r2如上所定义。
[0052]
具体步骤为：
[0053]
将化合物a与唑类试剂置于含有磁力转子的反应瓶中，加入甲苯作为溶剂和固体碱作为缚酸剂，加热搅拌反应，反应结束后，经冷却、萃取，减压蒸馏、柱层析得中间体b；将中间体b和三甲基碘化亚砜置于含有甲苯的反应瓶中，加入氢氧化钠溶液并加热反应，反应结束后，经冷却、萃取，有机相洗涤，减压除去溶剂、柱层析得中间体c；将中间体c溶解于甲醇中，加入叠氮化钠和氯化铵，搅拌反应，反应结束后，减压除去溶剂、柱层析得叠氮中间体d；将中间体d溶于异丙醇溶液，加入钯/碳，室温反应，反应结束后，减压除去溶剂、柱层析得关键中间体e；将中间体e和有机羧酸溶于干燥的dmf中，加入偶联试剂1-羟基苯并三唑和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐，室温反应，反应结束后，减压除去溶剂，经萃取，有机层洗涤、干燥，过滤，柱层析分离得纯净的目标产物。
[0054]
本发明优点：本发明采用药效基团拼接方式设计并合成了一类具有很强cyp51抑制活性的新型芳香丙烯酰胺-三氮唑类化合物，大部分的化合物具有显著的抗真菌活性，并且对耐药菌有较好的活性，体外细胞抑制活性明显优于氟康唑、咪康唑等药物。本发明提供的化合物代表了一类具有进一步开发价值的新结构类型的唑类cyp51抑制剂，为研发抗真菌新药和解决真菌耐药问题提供了基础。目标化合物的制备工艺简便，操作简单，总收率可达25％以上，适合工业化生产。
具体实施方式
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为对本发明进行更好地说明，举实施例如下：
[0056][0057]
实施例1 n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)丙烯酰胺的制备
[0058]
(1)1-(2,4-二氟苯基)-2-(1h-1,2,4-三氮唑-1-基)乙酮的制备
[0059]
称取2-溴-2',4'-二氟苯乙酮(5.0g)和1,2,4-三氮唑(2.22g)，并将其置于装有碳酸氢钠(2.68g)和甲苯(25ml)的100毫升圆底烧瓶中，回流状态下搅拌反应至原料消失，加入去离子水20ml,分液萃取，有机层用饱和食盐水洗涤、硫酸钠干燥，过滤除去干燥剂，减压除去溶剂，柱层析得目标产物(4.1g,收率86.4％)。1h nmr(400mhz,cdcl3)δ8.54(s,1h),8.07(dd,j＝15.0,8.1hz,2h),7.13
–
7.03(m,1h),7.03
–
6.92(m,1h),5.66(s,2h).
[0060]
(2)1-((2-(2,4-二氟苯基)环氧乙烷-2-基)甲基)-1h-1,2,4-三氮唑的制备
[0061]
将1-(2,4-二氟苯基)-2-(1h-1,2,4-三氮唑-1-基)乙酮(2.56g)和三甲基碘化亚砜(2.95g)溶于40ml甲苯中，搅拌下滴加氢氧化钠的水溶液(2.7ml,20％w/w)，加热至60℃，搅拌反应6小时。分备用水和饱和食盐水洗涤有机相，硫酸钠干燥，过滤除去干燥剂，减压除去溶剂，柱层析得目标产物(2.2g,收率80.9％)。1h nmr(400mhz,cdcl3)δ8.07(s,1h),7.88(s,1h),7.19(td,j＝8.4,6.5hz,1h),6.83(tdd,j＝10.6,8.3,2.4hz,2h),4.84(d,j＝14.9hz,1h),4.52(d,j＝14.9hz,1h),2.95(d,j＝4.7hz,1h),2.89(d,j＝4.7hz,1h).
[0062]
(3)1-氨基-2-(2,4-二氟苯基)-3-(1h-1,2,4-三氮唑-1-基)丙烷-2-醇的制备
[0063]
称取1-(2-(2,4-二氟苯基)环氧乙烷-2-基)甲基)-1h-1,2,4-三氮唑(1.0g)置于装有磁力搅拌子的圆底烧瓶中，相继加入甲醇(12ml)、叠氮化钠(0.82g)和氯化铵(0.29g)，加热至70℃，搅拌反应至原料消失。减压除去甲醇，加入二氯甲烷和水进行分液，水层用二氯甲烷洗涤2次，合并有机相并用硫酸钠干燥，减压除去溶剂得叠氮产物，直接用于下一步。
[0064]
将上述粗产物溶于异丙醇(15ml)中，加入5％钯/碳(20mg),在氢气环境下室温反应过夜，用滤纸抽滤除去钯/碳,减压除去溶剂并柱层析纯化得目标产物e(0.75g)。1h nmr(400mhz,cdcl3)δ8.09(s,1h),7.84(s,1h),7.57(td,j＝8.9,6.6hz,1h),6.94
–
6.68(m,2h),4.71
–
4.52(m,2h),3.21(dd,j＝12.9,0.9hz,1h),2.99(dd,j＝12.9,1.2hz,1h)；
13
c nmr(101mhz,cdcl3)δ164.16,163.81,161.80,161.67,160.47,157.82,151.70,144.78,130.51,130.45,130.42,130.36,124.72,111.86,111.82,111.65,111.62,104.70,104.44,104.17,74.43,74.38,55.98,55.92,47.24,47.19.
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(4)精确称量胺类中间体e(85.0mg,0.325mmol)与丙烯酸(49.8mg,0.25mmol)置于干燥的10ml双口圆底反应瓶中，加入无水dmf(10ml)，在氮气条件下加入1-羟基苯并三唑(hobt,26.0mg)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(edci,37.0mg)，氮气保护
下室温搅拌反应至羧酸消失。减压除去dmf，在反应瓶中加入乙酸乙酯(30ml)和稀氢氧化钠溶液(0.5mol/l,10ml)，萃取分液，用纯净水和饱和食盐水分别洗涤乙酸乙酯层，用无水硫酸镁对有机相进行干燥，过滤除去干燥剂并减压蒸馏除去乙酸乙酯，残渣进行柱层析纯化得纯品产物，收率67％。1h nmr(400mhz,dmso)δ8.27(s,1h),8.18(t,j＝5.9hz,1h),7.70(s,1h),7.30(dd,j＝8.9,7.0hz,1h),7.13(ddd,j＝11.9,9.2,2.5hz,1h),6.90(td,j＝8.5,2.4hz,1h),6.25
–
6.12(m,2h),5.99(dd,j＝17.1,2.1hz,1h),5.52(dd,j＝10.1,2.1hz,1h),4.58(d,j＝14.3hz,1h),4.46(d,j＝14.3hz,1h),3.61(d,j＝5.8hz,2h)；
13
c nmr(101mhz,cdcl3)δ168.05,164.41,164.30,162.87,161.94,161.83,160.14,160.02,157.69,157.57,151.61,144.89,130.65,130.59,130.55,130.50,129.58,128.38,123.93,123.80,112.15,112.12,111.95,111.92,104.62,104.36,104.09,99.68,76.58,76.53,56.11,56.06,47.53,47.48.
[0066]
实施例2
[0067]
n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)丙炔酰胺的制备
[0068]
按照实施例1的方法，收率63％。1h nmr(400mhz,dmso)δ8.44(t,j＝6.0hz,1h),8.30(s,1h),7.72(s,1h),7.33(dd,j＝16.0,9.0hz,1h),7.16(ddd,j＝11.8,9.2,2.4hz,1h),6.94(td,j＝8.6,2.5hz,1h),5.99(s,1h),4.61(d,j＝14.4hz,1h),4.49(d,j＝14.4hz,1h),3.64(dd,j＝14.5,6.2hz,1h),3.53(dd,j＝13.9,5.8hz,1h)；
13
c nmr(101mhz,dmso)δ169.62,163.13,160.69,160.57,160.39,157.81,157.70,153.32,150.62,150.52,144.94,130.17,130.10,130.07,130.01,124.48,124.44,124.35,124.31,110.79,110.59,104.16,103.90,103.62,83.56,75.21,74.61,74.55,54.94,54.89,45.91,45.86.
[0069]
实施例3
[0070]
n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)肉桂酰胺的制备
[0071]
按照实施例1的方法，收率73％。1h nmr(400mhz,dmso)δ8.33(s,1h),8.25(t,j＝5.8hz,1h),7.76(s,1h),7.54(d,j＝6.6hz,2h),7.46
–
7.30(m,5h),7.20(ddd,j＝11.7,9.3,2.4hz,1h),6.95(td,j＝8.5,2.2hz,1h),6.67(d,j＝15.8hz,1h),6.34(s,1h),4.65(d,j＝14.4hz,1h),4.54(d,j＝14.3hz,1h),3.73(d,j＝5.9hz,2h)；
13
c nmr(101mhz,dmso)δ170.33,166.38,163.12,160.68,160.56,160.31,160.19,157.86,157.73,150.56,144.97,139.37,134.69,130.28,130.19,130.12,129.58,128.92,127.58,124.79,124.66,121.46,110.86,110.66,104.21,103.93,103.67,74.98,74.93,55.19,46.27.
[0072]
实施例4
[0073]
n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)吡啶酰胺的制备
[0074]
按照实施例1的方法，收率71％。1h nmr(400mhz,dmso)δ8.78
–
8.56(m,2h),8.33(s,1h),8.08
–
7.91(m,2h),7.76(s,1h),7.68
–
7.54(m,1h),7.48
–
7.32(m,1h),7.18(ddd,j＝11.8,9.2,2.4hz,1h),6.92(td,j＝8.5,2.5hz,1h),6.34(s,1h),4.69(d,j＝14.4hz,1h),4.57(d,j＝14.3hz,1h),3.93(dd,j＝13.9,6.9hz,1h),3.81(dd,j＝13.8,5.7hz,1h)；
13
c nmr(101mhz,cdcl3)δ166.84,164.32,164.19,161.83,161.71,160.18,160.06,
157.73,157.61,151.59,148.79,148.45,144.74,137.60,130.60,130.54,130.51,130.45,126.83,123.96,123.93,123.83,123.79,122.54,111.98,111.95,111.77,111.74,104.57,104.32,104.30,104.05,76.61,76.56,56.08,56.03,47.48,47.43.
[0075]
实施例5
[0076]
n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)异烟酸酰胺的制备
[0077]
按照实施例1的方法，收率72％。1h nmr(400mhz,dmso)δ8.85
–
8.63(m,3h),8.34(s,1h),7.76(s,1h),7.65(d,j＝6.0hz,2h),7.40(dd,j＝15.9,8.9hz,1h),7.19(ddd,j＝11.8,9.3,2.4hz,1h),6.93(td,j＝8.5,2.3hz,1h),6.15(s,1h),4.73(d,j＝14.4hz,1h),4.60(d,j＝14.4hz,1h),3.79(ddd,j＝19.5,13.8,6.2hz,2h)；
13
c nmr(101mhz,cdcl3)δ167.38,164.41,162.04,161.92,160.16,157.71,157.59,152.09,150.78,144.83,140.78,130.39,130.33,130.30,130.24,124.62,124.12,123.51,123.42,123.38,121.00,112.28,112.25,112.08,112.04,104.83,104.56,104.30,76.54,76.49,55.63,55.58,47.34,47.30.
[0078]
实施例6
[0079]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(吡啶-2-基)丙烯酰胺的制备
[0080]
按照实施例1的方法，收率61％。1h nmr(400mhz,dmso)δ8.58(d,j＝4.1hz,1h),8.45(t,j＝5.8hz,1h),8.32(s,1h),7.82(td,j＝7.7,1.7hz,1h),7.74(s,1h),7.55(d,j＝7.8hz,1h),7.38(ddd,j＝14.9,12.6,5.1hz,3h),7.18(ddd,j＝11.8,9.2,2.4hz,1h),7.07(d,j＝15.4hz,1h),6.94(td,j＝8.5,2.4hz,1h),6.31(s,1h),4.65(d,j＝14.4hz,1h),4.54(d,j＝14.3hz,1h),3.71(d,j＝5.9hz,2h)；
13
c nmr(101mhz,cdcl3)δ168.08,152.79,151.82,150.23,144.89,141.24,137.18,130.61,125.23,124.50,123.28,112.14,111.97,104.39,104.12,103.98,76.68,76.63,56.17,56.13,47.80,47.76.
[0081]
实施例7
[0082]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(吡啶-3-基)丙烯酰胺的制备
[0083]
按照实施例1的方法，收率65％。1h nmr(400mhz,dmso)δ8.74(s,1h),8.55(d,j＝3.5hz,1h),8.32(s,1h),8.26(t,j＝5.9hz,1h),7.96(d,j＝8.0hz,1h),7.75(s,1h),7.41(ddd,j＝24.6,11.8,7.0hz,3h),7.19(ddd,j＝11.9,9.3,2.5hz,1h),6.94(td,j＝8.5,2.4hz,1h),6.78(d,j＝15.9hz,1h),6.28(s,1h),4.64(d,j＝14.4hz,1h),4.53(d,j＝14.4hz,1h),3.72(d,j＝5.9hz,2h)；
13
c nmr(101mhz,cdcl3)δ167.52,164.35,164.27,161.98,161.93,161.86,161.78,160.20,160.11,160.08,160.04,157.60,151.88,150.87,149.41,144.93,139.29,134.81,130.67,130.61,130.57,130.51,130.41,123.98,121.32,112.21,112.04,104.69,104.44,104.17,76.66,76.61,56.08,56.03,47.67,47.63.
[0084]
实施例8
[0085]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(吡啶-4-基)丙烯酰胺的制备
[0086]
按照实施例1的方法，收率61％。1h nmr(400mhz,dmso)δ8.61(d,j＝5.8hz,2h),
8.39
–
8.27(m,2h),7.76(s,1h),7.51(d,j＝6.0hz,2h),7.42
–
7.33(m,2h),7.20(ddd,j＝11.8,9.2,2.5hz,1h),6.98
–
6.86(m,2h),6.25(s,1h),4.65(d,j＝14.4hz,1h),4.54(d,j＝14.3hz,1h),3.78
–
3.67(m,2h)；
13
c nmr(101mhz,dmso)δ165.35,163.13,163.01,160.69,160.57,160.34,160.22,157.88,157.76,150.57,150.04,144.96,142.31,136.63,130.28,130.18,130.12,126.30,124.63,124.54,121.76,110.88,110.65,104.22,103.94,103.68,74.81,74.76,55.12,55.07,46.12,46.07.
[0087]
实施例9
[0088]
n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(萘-2-基)丙烯酰胺的制备
[0089]
按照实施例1的方法，收率73％。1h nmr(400mhz,dmso)δ8.46
–
8.29(m,2h),8.26
–
8.10(m,2h),7.98(d,j＝7.9hz,2h),7.76(d,j＝9.7hz,2h),7.70
–
7.49(m,3h),7.42(dd,j＝15.9,9.0hz,1h),7.21(ddd,j＝11.8,9.2,2.5hz,1h),6.96(td,j＝8.5,2.4hz,1h),6.75(d,j＝15.6hz,1h),6.34(s,1h),4.69(d,j＝14.4hz,1h),4.58(d,j＝14.3hz,1h),3.77(qd,j＝14.0,6.0hz,2h).
[0090]
实施例10
[0091]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(4-吗啉苯基)丙烯酰胺的制备
[0092]
按照实施例1的方法，收率35％。1h nmr(400mhz,dmso)δ8.33(s,1h),8.12(d,j＝5.8hz,1h),7.76(d,j＝3.4hz,1h),7.40(d,j＝8.7hz,3h),7.31(d,j＝15.7hz,1h),7.24
–
7.15(m,1h),6.95(d,j＝8.8hz,3h),6.44(t,j＝7.8hz,2h),4.63(d,j＝14.4hz,1h),4.52(d,j＝14.3hz,1h),3.77
–
3.64(m,6h),3.22
–
3.12(m,4h).
[0093]
实施例11
[0094]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(1h-吲哚-3-基)丙烯酰胺的制备
[0095]
按照实施例1的方法，收率45％。1h nmr(400mhz,dmso)δ11.59(s,1h),8.51
–
8.28(m,2h),8.15(t,j＝5.9hz,1h),7.87(d,j＝7.8hz,1h),7.76(d,j＝5.5hz,1h),7.59(d,j＝15.8hz,1h),7.50
–
7.32(m,2h),7.29
–
7.08(m,3h),6.95(t,j＝8.6hz,1h),6.62(d,j＝15.8hz,1h),4.63(d,j＝14.2hz,1h),4.54(d,j＝14.2hz,1h),3.75
–
3.65(m,2h)；
13
c nmr(101mhz,cdcl3)δ170.28,164.40,162.72,161.80,160.94,160.18,160.06,159.49,157.74,157.61,150.96,144.78,137.32,136.87,130.84,130.75,129.25,125.40,124.16,124.03,123.57,121.67,120.44,113.93,113.50,112.11,111.94,104.57,104.31,104.04,76.63,76.58,56.59,56.55,48.11,48.07.
[0096]
实施例12
[0097]
n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹喔啉-2-基)丙烯酰胺的制备
[0098]
按照实施例1的方法，收率62％。1h nmr(400mhz,dmso)δ9.16(s,1h),8.54(t,j＝6.0hz,1h),8.33(s,1h),8.14
–
8.00(m,2h),7.95
–
7.80(m,2h),7.76(s,1h),7.67(d,j＝15.6hz,1h),7.39(dd,j＝11.2,7.4hz,2h),7.20(ddd,j＝11.8,9.2,2.4hz,1h),7.01
–
6.90(m,1h),6.23(s,1h),4.67(d,j＝14.4hz,1h),4.57(d,j＝14.4hz,1h),3.77(d,j＝6.0hz,
2h).
[0099]
实施例13
[0100]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹啉-2-基)丙烯酰胺的制备
[0101]
按照实施例1的方法，收率62％。1h nmr(400mhz,dmso)δ8.49(t,j＝5.9hz,1h),8.41(d,j＝8.5hz,1h),8.32(d,j＝4.7hz,1h),8.03
–
7.92(m,2h),7.87
–
7.70(m,3h),7.68
–
7.52(m,2h),7.40(dd,j＝15.9,8.9hz,1h),7.30
–
7.12(m,2h),7.00
–
6.88(m,1h),6.26(s,1h),4.67(d,j＝14.4hz,1h),4.56(d,j＝14.3hz,1h),3.75(d,j＝6.0hz,2h)；
13
c nmr(101mhz,cdcl3)δ168.16,164.34,164.22,162.79,161.86,161.74,160.16,160.01,157.68,157.54,153.01,151.56,148.30,144.85,141.43,137.08,130.69,130.63,130.60,130.54,130.31,129.67,128.29,127.76,127.40,124.86,123.95,123.91,123.81,123.78,121.64,112.04,111.84,104.55,104.28,104.02,76.60,76.55,56.22,56.17,47.84,47.79.
[0102]
实施例14
[0103]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-甲氧基喹啉-2-基)丙烯酰胺的制备
[0104]
按照实施例1的方法，收率67％。1h nmr(400mhz,cdcl3)δ8.34
–
8.05(m,3h),7.85(s,1h),7.72(d,j＝15.3hz,1h),7.66
–
7.50(m,2h),7.46(d,j＝9.1hz,1h),7.28(d,j＝11.4hz,1h),7.10(s,1h),6.93
–
6.62(m,3h),6.10(s,1h),4.62(q,j＝14.0hz,2h),4.08
–
3.89(m,4h),3.83(dd,j＝14.0,5.0hz,1h)；
13
c nmr(101mhz,cdcl3)δ168.17,164.40,164.29,161.92,161.80,160.15,160.03,158.75,157.70,157.58,151.74,150.36,144.89,144.17,141.28,135.93,130.94,130.73,130.67,130.64,130.58,129.61,123.95,123.85,123.82,123.62,122.21,118.80,112.15,112.12,111.94,111.91,110.80,105.13,104.61,104.35,104.09,76.66,76.61,56.22,56.18,55.82,47.87,47.83.
[0105]
实施例15
[0106]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-甲基喹啉-2-基)丙烯酰胺的制备
[0107]
按照实施例1的方法，收率71％。1h nmr(400mhz,cdcl3)δ8.12(d,j＝6.1hz,2h),8.04(d,j＝8.8hz,1h),7.85(s,1h),7.72(d,j＝15.3hz,1h),7.60(d,j＝18.0hz,3h),7.49(d,j＝8.4hz,1h),7.14(d,j＝15.3hz,1h),6.89
–
6.74(m,2h),6.67(s,1h),6.13(s,1h),4.62(q,j＝14.1hz,2h),3.96(dd,j＝14.4,5.8hz,1h),3.82(dd,j＝14.4,5.5hz,1h),2.55(s,3h)；
13
c nmr(101mhz,cdcl3)δ168.06,164.39,164.28,161.91,161.80,160.15,160.02,157.69,157.57,151.95,151.72,146.60,144.89,141.22,137.78,136.72,132.94,130.71,130.66,130.62,130.56,129.09,128.44,126.64,124.69,123.96,123.82,121.82,112.11,111.94,111.91,104.61,104.34,104.08,100.19,76.65,76.60,56.20,56.16,47.85,47.80,21.85.
[0108]
实施例16
[0109]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-氟喹啉-2-基)丙烯酰胺的制备
[0110]
按照实施例1的方法，收率66％。1h nmr(400mhz,cdcl3)δ8.28
–
8.01(m,3h),7.86(s,1h),7.71(d,j＝15.1hz,1h),7.66
–
7.49(m,3h),7.45(d,j＝8.4hz,1h),7.08(d,j＝15.2hz,1h),6.90
–
6.71(m,2h),6.55(s,1h),4.62(q,j＝14.1hz,2h),3.97(dd,j＝14.3,5.9hz,1h),3.83(dd,j＝14.4,5.7hz,1h)；
13
c nmr(101mhz,cdcl3)δ167.65,162.46,159.96,157.72,157.60,152.21,152.18,151.77,144.88,144.69,140.30,137.33,137.27,131.77,131.68,131.12,130.68,130.62,130.59,130.53,129.12,129.06,129.02,125.70,123.89,123.85,123.75,123.72,122.57,121.45,121.20,112.19,111.98,111.11,110.89,104.68,104.41,104.15,76.65,76.60,56.14,56.10,47.82,47.78,30.79.
[0111]
实施例17
[0112]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-(三氟甲基)喹啉-2-基)丙烯酰胺的制备
[0113]
按照实施例1的方法，收率64％。1h nmr(400mhz,dmso)δ8.62(d,j＝8.6hz,1h),8.58
–
8.48(m,2h),8.33(s,1h),8.17(d,j＝8.9hz,1h),8.02(dd,j＝8.9,1.8hz,1h),7.91(d,j＝8.5hz,1h),7.76(s,1h),7.60(d,j＝15.6hz,1h),7.40(dd,j＝15.9,8.9hz,1h),7.31(d,j＝15.6hz,1h),7.21(ddd,j＝11.8,9.2,2.4hz,1h),6.96(td,j＝8.5,2.4hz,1h),6.25(s,1h),4.67(d,j＝14.4hz,1h),4.57(d,j＝14.3hz,1h),3.76(d,j＝5.7hz,2h)；
13
c nmr(101mhz,dmso)δ165.47,163.10,163.00,160.67,160.55,160.32,160.22,157.89,157.77,155.94,150.54,148.51,144.93,138.28,130.47,130.27,130.20,130.15,130.11,128.35,126.88,126.68,126.56,126.35,126.25,126.21,125.43,125.35,124.67,124.64,124.54,124.50,122.72,122.56,110.82,110.65,104.18,103.90,103.64,74.87,74.82,55.07,46.17.
[0114]
实施例18
[0115]
(e)-3-(6-氯喹啉-2-基)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)丙烯酰胺的制备
[0116]
按照实施例1的方法，收率68％。1h nmr(400mhz,dmso)δ8.51(t,j＝5.9hz,1h),8.39(d,j＝8.6hz,1h),8.33(s,1h),8.13(d,j＝2.3hz,1h),8.00(d,j＝9.0hz,1h),7.89
–
7.72(m,3h),7.56(d,j＝15.6hz,1h),7.40(d,j＝7.0hz,1h),7.31
–
7.12(m,2h),7.03
–
6.88(m,1h),6.26(s,1h),4.67(d,j＝14.4hz,1h),4.57(d,j＝14.3hz,1h),3.76(d,j＝5.9hz,2h)；
13
c nmr(101mhz,dmso)δ165.64,163.13,163.01,160.69,160.56,160.36,160.23,157.90,157.77,154.01,150.56,145.95,144.96,138.54,136.41,131.31,131.01,130.68,130.28,130.22,130.19,130.12,128.32,127.39,126.61,124.70,124.66,124.57,124.53,122.17,110.88,110.67,104.21,103.95,103.67,74.91,74.85,55.14,55.09,46.23,46.19.
[0117]
实施例19
[0118]
(e)-乙基2-(3-((2-2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)胺基)-3-氧丙基-1-烯-1-基喹啉-6-羧酸酯
[0119]
按照实施例1的方法，收率59％。1h nmr(400mhz,cdcl3)δ8.56(d,j＝1.6hz,1h),8.35
–
8.24(m,2h),8.14
–
8.02(m,2h),7.87(s,1h),7.73(d,j＝15.2hz,1h),7.61(dd,j＝15.7,9.1hz,1h),7.54(d,j＝8.5hz,1h),7.08(d,j＝15.2hz,1h),6.90
–
6.76(m,2h),6.36
(t,j＝6.0hz,1h),6.03(s,1h),4.71
–
4.56(m,2h),4.47(td,j＝14.2,7.1hz,2h),3.99(dd,j＝14.4,6.2hz,1h),3.82(dd,j＝14.7,5.8hz,1h),1.45(t,j＝7.1hz,3h)；
13
c nmr(101mhz,cdcl3)δ167.63,166.17,161.96,161.89,160.16,160.04,157.71,157.59,154.98,151.90,151.51,150.17,145.02,144.90,141.17,139.28,138.45,130.78,130.69,130.64,130.60,130.54,130.01,129.87,129.16,127.48,125.81,123.90,123.86,123.76,123.72,122.56,112.24,112.04,104.70,104.43,104.17,76.69,76.64,61.69,56.06,55.92,47.79,47.74,14.57.
[0120]
实施例20
[0121]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-苯基喹啉-2-基)丙烯酰胺的制备
[0122]
按照实施例1的方法，收率61％。1h nmr(400mhz,dmso)δ8.51(t,j＝6.0hz,1h),8.46(d,j＝8.5hz,1h),8.34(s,1h),8.29(d,j＝1.8hz,1h),8.13(dd,j＝8.9,2.0hz,1h),8.06(d,j＝8.8hz,1h),7.85(d,j＝7.3hz,2h),7.81
–
7.73(m,2h),7.56(dd,j＝21.1,11.7hz,3h),7.41(dt,j＝17.8,8.2hz,2h),7.21(dd,j＝7.4,4.7hz,1h),6.96(td,j＝8.5,2.5hz,1h),6.28(s,1h),4.67(d,j＝14.4hz,1h),4.57(d,j＝14.3hz,1h),3.76(d,j＝6.0hz,2h)；
13
c nmr(101mhz,dmso)δ165.83,163.15,163.02,162.30,160.70,160.58,160.37,160.25,157.91,157.79,153.49,150.57,146.97,144.97,139.12,138.93,138.41,137.30,130.23,129.54,129.28,129.12,128.01,127.92,127.06,126.86,125.20,124.70,124.57,121.65,110.87,110.69,104.22,103.96,103.68,74.95,74.90,55.16,46.24.
[0123]
实施例21
[0124]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(8-甲氧基-喹啉-2-基)丙烯酰胺的制备
[0125]
按照实施例1的方法，收率66％。1h nmr(400mhz,dmso)δ8.55(t,j＝5.9hz,1h),8.37
–
8.29(m,2h),7.84
–
7.69(m,2h),7.58(d,j＝15.6hz,1h),7.55
–
7.36(m,3h),7.27
–
7.12(m,3h),6.95(td,j＝8.5,2.4hz,1h),6.31(s,1h),5.75(s,1h),4.68(d,j＝14.3hz,1h),4.58(d,j＝14.3hz,1h),3.98(s,3h),3.77(d,j＝5.9hz,2h)；
13
c nmr(101mhz,dmso)δ166.04,163.15,163.03,160.71,160.59,160.37,160.25,157.91,157.79,155.21,152.00,150.56,144.98,139.50,139.26,136.73,130.30,130.24,130.21,130.15,128.67,127.44,126.43,124.75,124.72,124.62,124.59,121.49,119.29,110.89,110.66,108.89,104.20,103.94,103.67,79.15,75.01,74.96,55.69,55.21,55.16,46.35,46.31.
[0126]
实施例22
[0127]
(e)-3-(6-溴喹啉-2-基)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)丙烯酰胺的制备
[0128]
按照实施例1的方法，收率53％。1h nmr(400mhz,cdcl3)δ8.14
–
8.05(m,2h),7.98(d,j＝2.0hz,1h),7.93(d,j＝9.0hz,1h),7.87(s,1h),7.80(dd,j＝9.0,2.1hz,1h),7.71(d,j＝15.2hz,1h),7.66
–
7.55(m,1h),7.51(d,j＝8.5hz,1h),7.05(d,j＝15.3hz,1h),6.82(qd,j＝9.9,5.1hz,2h),6.32(t,j＝5.7hz,1h),6.03(s,1h),4.66(d,j＝14.2hz,1h),4.58(d,j＝14.2hz,1h),3.99(dd,j＝14.4,6.2hz,1h),3.82(dd,j＝15.1,6.0hz,1h)；
13
c nmr(101mhz,cdcl3)δ167.74,157.74,157.62,157.58,157.57,153.36,151.94,146.93,
144.95,141.17,136.28,136.23,134.00,131.44,130.73,130.67,130.63,130.58,129.99,129.92,129.39,125.30,123.90,123.81,122.72,121.59,112.27,112.24,112.07,112.04,104.47,76.72,76.67,56.15,47.77.
[0129]
实施例23
[0130]
((e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(6-碘喹啉-2-基)丙烯酰胺的制备
[0131]
按照实施例1的方法，收率41％。1h nmr(400mhz,cdcl3)δ8.21(d,j＝1.6hz,1h),8.10(s,1h),8.04(t,j＝9.4hz,1h),7.96(dd,j＝8.9,1.8hz,1h),7.86(d,j＝8.5hz,1h),7.73(dd,j＝22.9,12.1hz,2h),7.61(dd,j＝15.8,9.0hz,1h),7.49(d,j＝8.5hz,1h),7.03(d,j＝15.2hz,1h),6.89
–
6.72(m,2h),6.27(t,j＝5.7hz,1h),6.03(s,1h),4.66(d,j＝14.2hz,1h),4.57(d,j＝14.1hz,1h),3.99(dd,j＝14.3,6.3hz,1h),3.81(dd,j＝14.5,5.9hz,1h)；
13
c nmr(101mhz,cdcl3)δ167.50,153.29,151.70,150.13,147.12,144.68,141.05,138.92,136.47,136.40,135.65,131.17,130.51,130.46,130.41,130.33,130.30,129.68,124.97,122.32,112.04,112.00,111.83,111.79,104.48,104.21,93.02,76.47,76.42,55.84,47.56.
[0132]
实施例24
[0133]
(e)-n-(2-(2,4-二氯苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹啉-2-基)丙烯酰胺的制备
[0134]
按照实施例1的方法，收率57％。1h nmr(400mhz,dmso)δ8.32(t,j＝5.9hz,1h),8.24(d,j＝8.5hz,1h),8.12(d,j＝4.7hz,1h),7.93
–
7.85(m,2h),7.82
–
7.61(m,3h),7.62
–
7.50(m,2h),7.25(dd,j＝15.9,8.9hz,1h),7.18
–
7.12(m,2h),6.84
–
6.76(m,1h),6.21(s,1h),4.53(d,j＝14.4hz,1h),4.44(d,j＝14.3hz,1h),3.65(d,j＝6.0hz,2h)；
13
c nmr(101mhz,cdcl3)δ168.06,164.30,162.70,161.71,160.02,157.54,153.01,151.40,148.11,144.65,141.40,136.98,130.42,130.43,130.28,129.42,128.12,127.52,124.23,123.41,123.71,112.02,104.43,,76.52,56.12,47.67.
[0135]
实施例25
[0136]
(e)-n-(2-(2,4-二氟苯基)-2-羟基-3-(1h-1,2,4-三氮唑-1-基)丙基)-3-(喹啉-3-基)丙烯酰胺的制备
[0137]
按照实施例1的方法，收率65％。1h nmr(400mhz,dmso)δ9.09(s,1h),8.49(s,1h),8.30(dd,j＝13.3,7.8hz,2h),8.13
–
7.92(m,2h),7.86
–
7.71(m,2h),7.62(dd,j＝21.7,11.8hz,2h),7.41(dd,j＝16.0,8.7hz,1h),7.20(t,j＝9.7hz,1h),7.04
–
6.86(m,2h),6.31(s,1h),4.66(d,j＝14.3hz,1h),4.56(d,j＝14.3hz,1h),3.76(d,j＝5.6hz,2h)；
13
c nmr(101mhz,dmso)δ165.90,163.14,163.01,160.69,160.57,160.34,160.21,157.88,157.76,150.57,149.22,147.59,144.96,136.23,134.82,130.28,130.21,130.14,128.76,128.53,127.90,127.38,127.30,124.74,124.70,124.61,124.57,123.48,110.91,110.88,110.70,110.68,104.23,103.97,103.95,103.69,74.91,74.86,55.19,55.14,46.22,46.18.
[0138]
通过药理实验说明本发明酰胺-三氮唑类物质对真菌细胞的生长抑制作用。
[0139]
1.实验方法
[0140]
在96孔板上用bhi肉汤将目标化合物二倍稀释后加入到等体积生长对数期的菌液
中，目标化合物的浓度为1-64μg/ml，菌液细胞终浓度为1
×
106cfu/ml，孔溶液终体积为200μl。37℃厌氧环境中培育24h，酶标仪测定625nm下的od值。同时设置培养基空白对照、真菌对照以及氟康唑阳性药对照。
[0141]
mic
80
(％)＝(od样品
–
od空白)/(od参照
–
od空白)
×
100
[0142]
2.实验结果
[0143]
表一.酰胺-三氮唑类化合物类化合物对细菌生长抑制作用
[0144][0145]
注：5314为白色念珠菌；cg4为光滑念珠菌；h99为新型隐球菌
[0146]
通过对以上化合物的细菌细胞抑制活性评价结果可看出，大部分化合物均有一定的抑菌效果，化合物9、12、13、14、15、16、17、18、19、20、23对白色念珠菌、光滑念珠菌和新型
隐球菌生长抑制活性优于上市氟康唑、咪康唑、伊曲康唑等上市药物，具有明显的抑制效果，其中化合物9、13、16、18、20、23具有进一步开发研究价值。