CN117384136A - 一种no供体型抗真菌化合物及其制法和应用 - Google Patents
一种no供体型抗真菌化合物及其制法和应用 Download PDFInfo
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- CN117384136A CN117384136A CN202311270010.2A CN202311270010A CN117384136A CN 117384136 A CN117384136 A CN 117384136A CN 202311270010 A CN202311270010 A CN 202311270010A CN 117384136 A CN117384136 A CN 117384136A
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- Prior art keywords
- methyl
- difluorophenyl
- hydroxy
- arh
- triazol
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- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 40
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
- C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
- C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
Landscapes
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Abstract
本发明公开了一种NO供体型抗真菌化合物及其制法和应用,所述化合物为通式V所示,通过在活性唑类抗菌片段的基础上引入一氧化氮NO供体片段,到达在体外含巯基的亲核试剂存在下可剂量依赖性地缓慢释放一氧化氮的效果;上述化合物应用在抗真菌药物中,低浓度下即可抑制真菌生长,且具有广谱抗菌活性,对于白色念珠菌、光滑念珠菌和新型隐球菌有极强的抑制作用,同时,对于侵入性新型隐球菌带来的隐球菌性脑膜炎具有疗效。
Description
技术领域
本发明涉及一种抗真菌化合物,尤其涉及一种NO供体型抗真菌化合物,还涉及上述化合物的制法和应用。
背景技术
侵入性真菌感染(IFIs)日愈成为影响人类健康的重要因素(Science,2012,336(6082):647)。例如新生隐球菌作为全球范围内广泛存在的病原性真菌,能在人体内造成败血症、多器官衰竭和真菌性脑膜炎等众多危及生命的疾病(Nat.Rev.Drug.Discov.,2017,16(9):603-616;Fungal Genet.Biol.,2015,(78):16-48)。而引起IFIs致死率居高不下的原因之一是病原性真菌对抗真菌药物的耐受(Antimicrob.Agents Chemother.,2015,59(8):4982-4989;Antimicrob.Agents Chemother.,1999,43(8):1856-1861)。
在众多的耐药因素中,真菌生物被膜是不可忽视的。真菌生物被膜的产生为真菌提供了一个安全的庇护所,会阻挡抗生素的浸入并降解抗生素,生物被膜内细胞显示出更高水平的耐药性更强(Cell Chem.Biol.,2016,23(11):1383-1394)。由于其固有的耐药性,生物膜感染是一个正在出现的严重健康问题。生物膜是真菌致病过程中重要的毒力因子。一般来说,生物被膜被嵌入由多糖、蛋白质和微量eDNA组成的自产细胞外基质中(Antimicrob.Agents Chemother.2001,45(9),2475-2479;Nat.Rev.Drug.Disc.2003,2(2),114-122)。生物被膜的结构促进了养分和气体的交换。因此,真菌细胞能够抵抗环境、免疫细胞和药物的入侵。特别是,生物被膜中的细胞表现出与自由漂浮的浮游细胞截然不同的表型特征。因此,与生物被膜相关的感染本身就难以完全根除(Antimicrob.AgentsChemother.2001,45(9),2475-2479;Nat.Rev.Drug.Disc.2003,2(2),114-122)。据报道,真菌生物被膜的形成可以有效防御经典抗真菌药物如唑、两性霉素B等的入侵(Antimicrob.Agents Chemother.2002,46(11),3634-3636;Rev.Iberoam.Micol.2014,31(1),22-29)。由于唑类药物的广泛应用和长期的治疗方案,导致其由于生物被膜的耐药而面临潜在的治疗失败(Cold Spring Harbor Perspect.Med.2014,5(7),a019752;FutureMicrobiol.2014,9(4),523-542)。因此,迫切需要开发新的IFIs治疗策略,特别是克服与生物被膜相关的耐药性。
驱散生物被膜使真菌细胞重回浮游状态成为近年来备受关注的抗菌策略之一。然而,驱散后细胞重回自由状态的同时也存在着重新定殖、重新形成生物被膜的可能性,这会造成更加严重的二次感染,如何安全驱散真菌细胞被膜以及高效抑制真菌生长在抗真菌药物的研究中亟待解决。
发明内容
发明目的:本发明的目的是提供一种在释放NO的同时发挥唑类片段达到高效抑制真菌生长的NO供体型抗真菌化合物,并提供上述化合物的制法和应用。
技术方案:本发明的一种NO供体型抗真菌化合物如通式V所示:
其中,A为
当A为其中/>为4-羟基哌啶基/>二乙胺基D-脯氨醇基/>N-甲基哌嗪基/>四氢吡咯基/>或N-甲基乙醇胺基
当A为其中n=2,3,5;
当A为 X为脂肪烃或芳香烃。
优选的,当A为X为
优选的,当A为X为/> 或
优选的,所述化合物具体如下:
1a:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(4-羟基哌啶-1-基)二氮杂环形-1-鎓-1,2-二氢酸酯;
1b:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)胺-邻)-2,4-二硝基苯氧基-1-(S)-2-羟甲基吡咯烷-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯;
1c:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(吡咯烷-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯;
1d:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)氨基-邻)-2,4-二硝基苯;
1e:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(4-甲基哌嗪-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯;
1f:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)胺邻)-2,4-二硝基苯氧基-3-(2-羟乙基)-3-甲基二氮杂环戊烯-1-鎓-1,2-二氢酸酯;
2a:4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苄基)氨基)乙氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物;
2b:4-(3-(4-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苄基)氨基丙氧基)-3-苯磺酰基-1,2,5-恶二唑2-氧化物;
2c:4-((5-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基氨基)甲基)苄基)氨基)戊基)氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物;
2d:4-(4-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基)(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)丁氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物;
2e:4-(5-(2-(4-(3-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基)(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)戊基)氧基)-3-(苯基磺酰基)-1,2,5-恶二氮-2-氧化物;
2f:4-(2-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氨基)乙氧基)-3-(苯磺基尼龙基)-1,2,5-恶二唑2-氧化物;
2g:4-(2-(2-(4-(3-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氨基)乙氧基)-3-(仅苯基磺)-1,2,5-氧杂二氮唑-2-氧化物;
2h:4-(4-(4-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)丁-2-炔基-1-基)氧基)-3-(PH-烯基磺酰基)-1,2,5-恶二唑2-氧化物;
2i:4-(2-(2-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)乙氧基)乙氧基)-3-(苯丙氨酸-尼龙磺酰基)-1,2,5-恶二唑2-氧化物;
3a:2-(二羟氧基)乙基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3b:3-(硝氧基)丙基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3c:4-(硝氧基)丁基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3d:5-(硝氧基)戊基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三偶氮-l-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3e:4-((硝氧基)甲基)苄基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3f:2-((硝氧基)甲基)苄基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3g:(3S,3aR,6R,6aS)-6-(硝氧基)六氢呋喃[3,2-b]呋喃-3-基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)氨基)甲基-基苯氧基)丙氧基)氢烯基)乙酸酯;
3h:2-(2-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰基)-L2-氮杂基)乙基硝酸酯。
上述化合物的制法,具体如下:以仲胺类化合物4a-f为原料,高压下与NO气体反应得到偶氮鎓二醇钠盐类化合物5a-f,在过弱碱的作用下与2,4-二硝基氟苯发生亲核取代反应生成化合物6a-f;同时,取三甲基碘化亚砜在强碱环境加热生成碳负离子中间体C-H3,再与原料Az-1发生环氧化反应得到Az-2,Az-2在碱性条件下被胺类化合物开环得到Az-3,最后化合物6a-f在Az-3的碱性进攻下制得目标化合物;
上述化合物的制法,具体如下:以苯硫酚钠7为原料,与氯乙酸反应得到苯硫基乙酸8,然后8氧化得到化合物9,除去溶剂后在高温下与发烟硝酸反应得到呋咱氮氧化物10,呋咱氮氧化物在碱性条件下与Boc保护的氨基醇反应得到11a-c,脱保护后即得到中间体12a-c;同时,中间体Az-3与4-溴甲基苯甲醛发生亲核加成反应得到Az-4;最后,12a-c和Az-4通过弱还原剂腈基硼氢化钠的还原胺化得到目标化合物;
上述化合物的制法,具体如下:以对羟基苯乙酸甲酯14为原料与1,3-二溴丙烷在碱性条件下发生亲核加成得到15,15进一步与对羟基苯甲醛反应得到16,纯化后与甲胺盐酸盐在无水条件、三乙胺的催化下得到亚胺中间体,直接还原得到化合物17;将Az-2溶于乙腈中并加入过量三乙胺游离,后加入17在碱性环境下开环得到化合物Az-5,脱去乙酯部分即得到中间体Az-6;同时,由10与二醇在碱性环境下发生亲核取代得到13d-h;最后,Az-6和13d-h在催化剂的作用下缩合得到目标化合物;
上述化合物的制法,具体如下:以端基溴醇为原料,在高温避光条件下与硝酸银反应生成羟基硝酸酯类化合物,后与Az-6一步酯缩合得到目标化合物;
本发明还公开了一种药物组合物,由通式V的化合物或其医学上可接受的盐与药学上可接受的载体或辅料的药物组成。
上述的化合物在制备抗真菌的药物中的应用。
发明原理:本发明的NO供体型抗真菌化合物,通过在活性唑类抗菌片段的基础上引入一氧化氮NO供体片段,到达在体外含巯基的亲核试剂存在下可剂量依赖性地缓慢释放NO的效果,NO可通过降低胞内第二信使环二鸟苷酸(c-di-GMP)水平抑制真菌生物被膜的形成与促使生物被膜的驱散,进而影响真菌的生长;化合物稳定性较高,通过将化合物3a中的酯键替换为酰胺键,即化合物3h,显著提高了化合物3h在大鼠血浆和肝微粒中的稳定性。
NO供体型抗真菌化合物3a-3h通过穿透真菌细胞,破坏真菌细胞膜的完整性,在体内缓慢释放并产生有效浓度的NO,发挥显著的抗真菌活性,达到有效抑制新生隐球菌生长的效果,并且具有广谱抗菌活性;其中,化合物3a由于其一氧化氮供体部分空间结构更小而更有利于与CYP51(甾醇14α-去甲基化酶)相结合,低浓度下也能高效抑制新生隐球菌生长,通过抑制新生隐球菌生物膜形成,远优于阳性药氟康唑(FLC),也可以抑制新生隐球菌麦角甾醇的生物合成,从而发挥抗菌功能。化合物3b可以有效抑制新生隐球菌生长,优于FLC;化合物3e,3g能有效抑制白念珠菌生长。同时,滴鼻给予化合物3a和3h,可以显著降低小鼠脑组织和肺组织荷菌量,化合物3a和3h可显著减轻小鼠脑组织病理损伤程度。
有益效果:与现有技术相比,本发明具有如下显著优点:
(1)本发明的NO供体型唑类衍生物在释放NO的同时发挥唑类片段的抗真菌作用,达到发挥多个靶点之间的协同作用的效果,避免单靶点药物联合使用时因药代动力学性质复杂而造成的治疗窗变窄和给药剂量难以设置等问题;
(2)通过体外抗真菌活性测试,筛选出高活性的硝酸酯唑类衍生物3a和3e,其对新生隐球菌H99具有优秀的体外抑制活性,可以在﹤1μg/mL的低浓度时即可达到极高的抗隐球菌活性,显著优于现有抗真菌药物;
(3)硝酸酯类NO供体型唑类衍生物同时能够抑制新生隐球菌细胞膜上的麦角甾醇的生成从而发挥抗真菌活性;
(4)由于本申请的化合物优越的体内外抗新生隐球菌活性,可作为一类治疗隐球菌性脑膜炎的抗真菌新骨架,应对侵袭性真菌感染。
附图说明
图1为本发明NO供体型抗真菌化合物的通式V;
图2为化合物1f、2i、3a、3h的体外一氧化氮释放实验结果;
图3为化合物2a-2i和3a-3f的体外抗真菌活性研究以及化合物3a和3e的平皿扩散实验;
图4为化合物3a、3e在体外的抗真菌生物膜形成作用;
图5为化合物3a、3e在体内的NO释放实验;
图6为化合物3a、3e对新型隐球菌细胞麦角甾醇生物合成及微观结构的影响;
图7为化合物3a、3h的体内抗真菌活性研究。
具体实施方式
下面结合附图对本发明的技术方案作进一步说明。
实施例1
-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(4-羟基哌啶-1-基)二氮杂环形-1-鎓-1,2-二氢酸酯(1a)的制备
(a)将4-羟基哌啶(0.72g,5mmol)、新制的甲醇钠/甲醇溶液中(30%w/w,100mL)、无水乙醚(300mL)先后加入制特质高压反应釜中,将空气用氮气置换出后,通入NO气体,室温机械搅拌,反应72h后,加入大量乙醚析出固体,抽滤后使用无水乙醚洗涤,室温真空干燥得产物5a,不经纯化直接投入下一步反应。
(b)在氮气保护下将1,5-二氟-2,4-二硝基苯(204mg,1mmol)在15mL丙酮中的溶液冷却至0℃。向其中滴加偶氮鎓二醇钠盐5a(1.1mmol)在15mL 5%碳酸氢钠水溶液中的溶液。反应完成后,减压浓缩除去丙酮,将残余物吸收在CH2Cl2中并用水洗涤。有机溶液用硫酸钠干燥并真空蒸发得到所需产物,将其从乙醇中重结晶,得到化合物6a,淡黄色固体,收率35%。1H NMR(500MHz,Chloroform-d)δ8.91(d,J=7.5Hz,1H,ArH),7.43(d,J=11.7Hz,1H,ArH),4.06-4.00(m,1H,CH),3.92-3.86(m,2H,CH2),3.69-3.63(m,2H,CH2),2.09-2.02(m,2H,CH2),1.88-1.81(m,2H,CH2).
(c)将2,4-二氟-2-[1-(1H-1,2,4-三唑基)]苯乙酮(7.30g,32.7mmol)加入适量甲苯(150mL)中,加入三甲基碘化亚砜(8.64g,39.30mmol),然后缓慢加入20%氢氧化钠溶液(8mL)。然后将反应混合物在45-50℃加热3小时。反应结束后,加入EA(50mL)稀释,有机层用水(2×200mL)、饱和氯化钠溶液(200mL)洗涤,用Na2SO4干燥并过滤。减压浓缩滤液得浅棕色油状物;将油状物加入EA(100mL)缓慢滴加甲磺酸,有白色固体生成。反应3h后,迅速抽滤并用EA洗涤,迅速旋干得白色固体即Az-2,收率60%。1H NMR(300MHz,Chloroform-d)δ8.05(s,1H,ArH),7.86(s,1H,ArH),7.22-7.11(m,1H,ArH),6.87-6.75(m,2H,ArH),4.81(d,J=14.8Hz,1H,CH2),4.50(d,J=14.9Hz,1H,CH2),2.90(dd,J=18.7,4.6Hz,2H,CH2).
(d)取Az-2(500mg,0.156mmol),加入乙醇后加入三乙胺搅拌溶清后,加入25%甲胺乙醇溶液130mg,再加入三乙胺426mg,回流反应4h,TLC检测反应完全后减压蒸馏去除溶剂,硅胶柱层析纯化(CH2Cl2/MeOH,15/1,v/v)旋干得到黄色油状液体,即Az-3,收率30%。1H-NMR(300MHz,Chloroform-d)8.22(s,1H,ArH),8.17(s,1H,ArH),7.70-7.82(m,1H,ArH),6.95-6.97(m,2H,ArH),4.50-4.55,4.53-4.68(dd,2H,CH2),2.76-2.80,3.11-3.15(dd,2H,CH2),2.28-2.30(s,3H,CH3).
(e)将6a(57mg,0.311mmol)加入THF(10mL)中搅拌溶清,加入Az-3(92mg,0.343mmol)使其与6a充分混匀,再加入Na2CO3(66mg,0.622mmol)室温反应6h,TLC检测反应完全后,减压蒸馏加入乙醇溶解完全,抽滤出碳酸钠后柱层析(CH2Cl2/MeOH,10/1,v/v)后得到黄色固体,即1a,收率40%。1H-NMR(CDCl3,300MHz)8.61(s,1H,ArH),7.92(s,1H,ArH),7.83(s,1H,ArH),7.43-7.51(m,1H,ArH),7.10(s,1H,ArH),6.69-6.82(m,2H,ArH),4.59-4.63,4.71-4.75(dd,2H,CH2),4.00-4.04(m,1H,CH),3.92(s,2H,NCH2),3.81-3.86(m,2H,CH2),3.57-3.64(m,2H,CH2),2.92(s,3H,NCH3),2.83(s,1H,OH),2.02-2.09(m,2H,CH2),1.81-1.86(m,2H,CH2).13C NMR(126MHz,CDCl3)δ164.24-164.14,162.23-162.13(dd,J=247.5,12Hz),159.53-159.44,157.59-157.49(dd,J=247.5,12Hz),153.05,151.90,151.69,150.42,150.14,149.97,144.59,144.45,133.07,131.05,131.01,130.32-130.20(dd,J=9,6Hz),130.10-129.98(dd,J=9,6Hz),129.35,127.87,127.12,122.98-122.88(d,J=12Hz),112.32-112.02(d,J=20.25Hz),108.75,105.99,104.44-104.02(t,J=26.25Hz),66.19,64.79,59.65,54.71,54.67,48.26,47.59-47.21(m,2C),45.06,44.17,31.94-31.74(m,2C).HRMS(ESI-MS m/z)calculated for C23H25F2N9O8[M+Na]+616.1669,Found 616.1675ppm(parts per million)error:0.9.
实施例2
5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)胺-邻)-2,4-二硝基苯氧基-1-(S)-2-羟甲基吡咯烷-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯(1b)的制备
参照实施例1的合成方法。1H NMR(CDCl3,500MHz)8.57(d,J=7.6Hz,1H,ArH),7.92(s,1H,ArH),7.83(s,1H,ArH),7.46(d,J=11.8Hz,1H,ArH),7.09-7.12(m,1H,ArH),6.69-6.82(m,2H,ArH),4.59-4.63,4.71-4.75(dd,2H,-CH2-),3.93-3.95(m,2H,NCH2-),3.88-3.91(m,2H,prolinol),3.84-3.86(m,1H,prolinol),3.73-3.79(m,2H,prolinol),2.84-2.86(s,3H,NCH3),2.03-2.17(m,4H,prolinol).13C NMR(126MHz,CDCl3)δ164.19-164.09,162.19-162.09(dd,J=247.5,12Hz),159.58-159.48,157.63-157.53(dd,J=247.5,12Hz),153.25,153.19,151.76,150.34,144.54,132.51,130.32-130.21(dd,J=9,6Hz),127.14(s,2C),123.03-122.95(d,J=12Hz),112.22-112.06(d,J=20.25Hz),105.03,104.40-103.98(t,J=26.25Hz),64.20(s,3C),59.52,54.85-54.81(d,1C),52.53-52.47(d,1C),45.04(s,2C),27.34-27.21(d,1C),23.21-23.14(d,1C).HRMS(ESI-MS m/z)calculated for C23H25F2N9O8[M+Na]+616.1669,Found 616.1673ppm error:0.8.
实施例3
5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(吡咯烷-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯(1c)的制备
参照实施例1的合成方法。1H NMR(DMSO-d6,300MHz)8.57(d,J=7.6Hz,1H,ArH),7.92(s,1H,ArH),7.83(s,1H,ArH),7.46(d,J=11.8Hz,1H,ArH),7.09-7.12(m,1H,ArH),6.69-6.82(m,2H,ArH),4.59-4.63,4.71-4.75(dd,2H,-CH2-),3.78-3.83(m,4H,pyrrolidine),3.77-3.82,4.06-4.18(dd,2H,-NCH2-),2.84-2.86(s,3H,NCH3),2.05-2.12(m,4H,pyrrolidine).HRMS(ESI-MS m/z)calculated for C22H23F2N9O7[M+H]+563.1689,found 564.1668ppm error:-0.5.
实施例4
5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)氨基-邻)-2,4-二硝基苯(1d)的制备
参照实施例1的合成方法。1H NMR(CDCl3,500MHz)8.57(s,1H,ArH),7.97(s,1H,ArH),7.82(s,1H,ArH),7.46(m,1H,ArH),7.09(s,1H,ArH),6.69-6.77(m,2H,ArH),4.59-4.63,4.71-4.75(dd,2H,-CH2-),3.93-3.95(m,2H,-NCH2-),3.50-3.51(d,4H,-NCH2CH3),2.93(s,3H,NCH3),1.22-1.24(t,6H,-NCH2CH3).13C NMR(75MHz,DMSO-d6)δ160.12,155.34,148.40,147.36,145.38,139.57,128.24,125.50(dd,J=15.0Hz),123.16,122.39,118.07(dd,J=15.0Hz),107.59,107.31,101.16,99.41(t,J=105.0Hz),54.80,49.75,48.76,42.39,40.28,6.81.HRMS(ESI-MS m/z)calculated for C22H25F2N9O7[M+Na]+588.1732,found 588.1737ppm error:0.59.
实施例5
5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(4-甲基哌嗪-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯(1e)的制备
参照实施例1的合成方法。1H NMR(CDCl3,500MHz)8.61(s,1H,ArH),7.90(s,1H,ArH),7.80(s,1H,ArH),7.46(m,1H,ArH),7.08(s,1H,ArH),6.69-6.80(m,2H,ArH),4.59-4.63,4.71-4.75(dd,2H,-CH2-),3.88-3.96(m,2H,-NCH2-),3.65-3.67(m,4H,piperazine),2.93(s,3H,-NCH3),2.64-2.66(m,4H,piperazine),2.37(s,3H,piperazine).13C NMR(75MHz,DMSO-d6)δ152.42,151.35,150.49,150.16,145.58(d,J=6.0Hz),145.05(d,J=6.0Hz),132.45,127.31(d,J=9.0Hz),126.87,111.69,111.42,106.23,104.22,76.56(d,J=9.0Hz),60.17,55.78,53.29,50.32,45.39,44.09.HRMS(ESI-MS m/z)calculated for C23H26F2N10O7[M+Na]+615.1836,found615.1846ppm error:1.56.
实施例6
5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)胺邻)-2,4-二硝基苯氧基-3-(2-羟乙基)-3-甲基二氮杂环戊烯-1-鎓-1,2-二氢酸酯(1f)的制备
参照实施例1的合成方法。1H NMR(CDCl3,500MHz),8.61(s,1H,ArH),7.90(s,1H,ArH),7.81(s,1H,ArH),7.46(m,1H,ArH),7.08(s,1H,ArH),6.69-6.77(m,2H,ArH),3.93-3.95(m,2H,-NHCH2-),3.88-3.90(t,2H,-NHCH2CH2OH),3.72-3.80(m,2H,-NHCH2CH2OH),3.34(s,3H,-NHCH3),2.84(s,3H,-NHCH3).13C NMR(75MHz,CDCl3)δ181.62,153.25,151.78(d,J=21.0Hz),150.23,144.51,132.80,130.09,127.62,127.17,122.93(d,J=69.0Hz),112.42(d,J=12.0Hz),112.14(d,J=12.0Hz),105.50,104.26(t,J=105.0Hz),54.96,59.34(d,J=15.0Hz),55.89,54.70(d,J=27.0Hz),54.39(d,J=24.0Hz),44.99,40.77.HRMS(ESI-MS m/z)calculated for C21H23F2N9O8[M+Na]+590.1523,found590.1529ppm error:0.91.
实施例7:
4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苄基)氨基)乙氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物(2a)的制备
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(a,b,c)将苯硫酚钠7溶入乙醇中,升温使其全部溶解后加入氯乙酸与碳酸钾回流反应,TLC检测反应,待反应完成后减压蒸馏加入1N HCl酸化,待pH值<2后又白色固体析出,将白色固体抽滤得到化合物8,化合物8溶于冰醋酸后加入过量30% H2O2,回流反应过夜后除去大部分溶剂,重新加入少量冰醋酸,将温度升至90℃后加入发烟硝酸,再将反应温度进一步升至120℃,反应2h后在反应液中倾入冰水,有白色固体析出,抽滤洗涤即得到化合物10,白色固体,收率36%。1H NMR(500MHz,Chloroform-d)δ8.17(d,J=8.5Hz,4H,ArH),7.80(d,J=7.4Hz,2H,ArH),7.70-7.63(m,4H,ArH).
(d)化合物10(500mg,1.366mmol)溶于THF中,待溶清后加入N-(叔丁氧羰基)氨基醇(2.732mmol),再缓慢滴加50% NaOH溶液(109mg,2.732mmol,以NaOH计),TLC(PE/EA,5/1,v/v)检测反应,结束后旋干溶剂通过硅胶柱层析纯化(PE/EA,10/1,v/v),得到白色固体化合物11a。
(e)将11a-c(100mg)溶于DCM中,将反应液冷浴至0℃后加入TFA,反应由TLC(PE/EA,3/1,v/v)检测,反应结束后旋干加入EA(20mL)使用饱和碳酸氢钠,纯水,饱和氯化钠洗涤,将有机层收集用Na2SO4干燥,抽滤,除去溶剂后得到化合物12a,白色固体,收率47%。1HNMR(500MHz,DMSO-d6)δ8.04(d,J=7.7Hz,2H,ArH),7.90(d,J=7.5Hz,1H,ArH),7.75(d,J=7.8Hz,2H,ArH),4.34(d,J=5.5Hz,2H,CH2),2.92(d,J=5.6Hz,2H,CH2).
(f)将Az-3(536mg,2mmol)溶于ACN(30mL)中待搅拌溶清后加入K2CO3(828mg,6mmol),在常温下反应4h反应液变浑浊,TLC(PE/EA,5/1,v/v)检测反应,反应完全后柱层析得到白色固体即化合物Az-4,收率74%。1H NMR(CDCl3,300MHz)8.22(s,1H,ArH),8.17(s,1H,ArH),7.70-7.82(m,1H,ArH),7.30-7.38(d,2H,ArH),7.14-7.19(d,2H,ArH),6.95-6.97(m,2H,ArH),4.50-4.55,4.53-4.68(dd,2H,-CH2-),3.76(s,2H,-NCH2-),2.76-2.80,3.11-3.15(dd,2H,NCH2-),2.25(s,3H,NCH3).
(g)将Az-4(100mg,0.373mmol)与12a(0.410mmol)共同溶于甲醇(15mL)中,在室温下反应2h,TLC(CH2Cl2/MeOH,20/1,v/v)检测反应,反应结束后分批缓慢加入腈基硼氢化钠(118mg,1.865mmol)反应过夜,TLC(CH2Cl2/MeOH/Et3N,10/1/1drop,v/v/v)检测结束后旋干溶剂,使用碱性氧化铝柱层析(CH2Cl2/MeOH/Et3N,10/1/1%,v/v/v),得到白色固体,即目标化合物2a,白色固体,收率:27%。1H NMR(CDCl3,500MHz)8.04-8.07(m,2H,ArH),7.75-7.78(m,2H,ArH),7.65-7.70(m,1H,ArH),7.60-7.63(m,2H,ArH),7.34-7.35(m,2H,ArH),7.14-7.15(m,2H,ArH),6.81-6.89(m,2H,ArH),4.68(s,J=6Hz,2H,NCH2CH2O-),4.32-4.35,4.51-4.55(dd,2H,-CH2-),4.05(s,2H,Ar-CH2-),3.33-3.36,3.54-3.56(dd,2H,NCH2-Ar),3.36(t,J=7Hz,2H,NCH2CH2O-),2.64-2.67,3.01-3.04(dd,2H,NCH2-),2.06(s,3H,NCH3).HRMS(ESI-MS m/z)calculated for C30H31F2N7O6S[M+H]+656.2097,found656.2102ppm error:-0.77.
实施例8
4-(3-(4-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苄基)氨基丙氧基)-3-苯磺酰基-1,2,5-恶二唑2-氧化物(2b)的制备
参照实施例7的合成方法。1H NMR(CDCl3,500MHz)8.06-8.07(m,2H,ArH),7.75-7.78(m,2H,ArH),7.62-7.65(m,3H,ArH),7.34-7.35(m,2H,ArH),7.14-7.15(m,2H,ArH),6.80-6.86(m,2H,ArH),4.57-4.60(t,J=6Hz,2H,CH2O-),4.32-4.35,4.51-4.55(dd,2H,-CH2-),4.05(s,2H,Ar-CH2-),3.33-3.36,3.54-3.56(dd,2H,NCH2-Ar),3.36(t,J=7Hz,2H,NCH2-),2.64-2.67,3.01-3.04(dd,2H,NCH2-),2.09-2.16(m,2H,-CH2CH2CH2-),2.06(s,3H,NCH3).HRMS(ESI-MS m/z)calculated for C31H34F2N7O6S[M+H]+670.2253,found670.2257ppm error:-0.44.
实施例9
4-((5-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基氨基)甲基)苄基)氨基)戊基)氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物(2c)的制备
参照实施例7的合成方法。1H NMR(CDCl3,500MHz)8.09-8.11(m,2H,ArH),7.88(m,1H,ArH),7.75-7.78(m,2H,ArH),7.73(m,1H,ArH),7.62-7.65(m,2H,ArH),7.34-7.35(m,2H,ArH),7.14-7.15(m,2H,ArH),6.96-6.99(m,1H,ArH),6.85-6.88(m,1H,ArH),4.57-4.60(t,J=6Hz,2H,CH2O-),4.32-4.35,4.51-4.55(dd,2H,-CH2-),4.05(s,2H,Ar-CH2-),3.33-3.36,3.54-3.56(dd,2H,NCH2-Ar),3.36(t,J=7Hz,2H,CH2O-),2.64-2.67,3.01-3.04(dd,2H,NCH2-),2.09-2.16(m,2H,-CH2-),2.06(s,3H,NCH3),1.99-2.02(m,2H,-CH2-),1.73-1.77(m,2H,-CH2-).HRMS(ESI-MS m/z)calculated for C33H37F2N7O6S[M+H]+698.2528,found 698.2439ppm error:1.27.
实施例10
4-(4-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基)(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)丁氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物(2d)的制备
(a)对羟基苯乙酸甲酯14(10g,60.2mmol)、1,3-二溴丙烷(12.1g,90.3mmol)、K2CO3(24.88g,180.3mmol)溶于无水乙醇(250mL)中,在85℃下反应过夜,TLC(PE/EA,8/1,v/v)检测反应,待反应完全后抽滤除去碳酸钾,旋蒸除去大部分溶剂,再加入乙酸乙酯(100mL),水(100mL)萃取3次并用饱和氯化钠溶液洗涤后合并有机层,加入无水Na2SO4干燥,抽滤,硅胶拌样柱层析(PE/EA,15/1,v/v)获得化合物15,淡黄色液体,收率53%。1H NMR(500MHz,Chloroform-d)δ7.19(d,J=8.4Hz,2H,ArH),6.86(d,J=8.4Hz,2H,ArH),4.09(d,J=5.5Hz,2H,CH2),3.69(s,3H,CH3),3.60(d,J=6.4Hz,2H,CH2),3.57(s,2H,CH2),2.34-2.28(m,2H,CH2).
(b)将化合物15(3g,10.4mmol)溶于无水乙醇(30mL)中,加入对羟基苯甲醛(1.9g,15.6mmol)与碳酸钾(4.3g,31.2mmol),在85℃下回流反应6h,旋干后加入EA(100mL)抽滤去除固体,使用水萃取3次饱和食盐水洗涤1次,合并有机层,以无水Na2SO4干燥1h,过滤,旋蒸除去溶剂,硅胶拌样后以(PE/EA,15/1,v/v)为流动相经硅胶柱层析纯化得到化合物16,白色固体,收率42%。1H NMR(500MHz,Chloroform-d)δ9.88(s,1H,CHO),7.83(d,J=8.7Hz,2H,ArH),7.19(d,J=8.7Hz,2H,ArH),7.01(d,J=8.6Hz,2H,ArH),6.87(d,J=8.7Hz,2H,ArH),4.25(t,J=6.2Hz,2H,CH2),4.15(d,J=6.0Hz,2H,CH2),3.68(s,3H,CH3),3.56(s,2H,CH2),2.29(p,2H,CH2).
(c)将化合物16(1g,2.9mmol),超声搅拌溶于无水甲醇(20mL)中,加入甲胺盐酸盐(398mg,5.8mmol)、三乙胺(879mg,8.7mmol)、无水Na2SO4在室温下搅拌反应过夜,TLC(CH2Cl2/MeOH,15/1,v/v)检测,加入硼氢化钠(812mg,11.6mmol),继续搅拌反应1h。TLC(CH2Cl2/MeOH/Et3N,10/1/1%,v/v/v)检测反应,旋干加入乙酸乙酯抽滤并用乙酸乙酯清洗滤饼,收集滤液,有机层用水萃取三次饱和食盐水洗涤一次后无水Na2SO4干燥,过滤,滤液经减压除去有机溶剂,残余物即为化合物17,白色固体,收率93%。1H NMR(500MHz,Chloroform-d)δ7.22(d,J=8.6Hz,2H,ArH),7.18(d,J=8.6Hz,2H,ArH),6.87(d,J=1.3Hz,2H,ArH),6.86(d,J=1.3Hz,2H,ArH),4.14(d,J=6.1Hz,4H,CH2),3.68(s,5H),3.56(s,2H,CH2),2.43(s,3H,CH3),2.24(p,J=6.1Hz,2H,CH2).
(d,e)将化合物Az-2(1.12g,3.4mmol)溶于乙腈(20mL)中,加入三乙胺(565mg,5.6mmol)待完全溶解后,加入17(1g,2.8mmol),在50℃反应4h,溶液变黄,TLC(CH2Cl2/MeOH,10/1,v/v)检测,反应结束后旋干,重新加入1,4-二氧六环稀释(20mL)再加入1N NaOH溶液搅拌脱去乙酯,溶液变浑浊,TLC(CH2Cl2/MeOH,10/1,v/v)检测结束后旋干,加入水(30mL)并用1N HCl调节pH值至7,有大量固体析出,用乙酸乙酯(100mL)萃取三次,合并有机层,用饱和食盐水洗涤一次,无水Na2SO4干燥,过滤,滤液减压除去有机溶剂,得到白色固体,乙醚打浆,过滤,柱层析,即中间体Az-6,白色固体,收率25%。1H NMR(500MHz,DMSO-d6)δ8.27(s,1H,ArH),7.73(s,1H,ArH),7.41(td,J=9.0,6.8Hz,1H,ArH),7.15(d,J=8.4Hz,2H,ArH),7.13-7.10(m,1H,ArH),6.99(d,J=8.5Hz,2H,ArH),6.95(td,J=8.5,2.6Hz,1H,ArH),6.88(d,J=8.7Hz,2H,ArH),6.82(d,J=8.6Hz,2H,ArH),4.56-4.44(m,2H,ArCH2),4.09(d,J=5.3Hz,4H,OCH2),3.47(s,2H,ArCH2),3.45(d,J=13.1Hz,1H,NCH2),3.33(d,J=13.2Hz,1H,CH2),2.96(dd,J=14.1,1.4Hz,1H,CH2),2.75(d,J=13.7Hz,1H,CH2),2.13(d,J=6.2Hz,2H,CH2),2.04(s,3H,CH3).
(f)将化合物10(500mg,136.6mmol)溶于THF中,加入二醇(273.2mmol),再加入50% NaOH(273.3mmol),室温反应3-5h,TLC(PE/EA,2/1,v/v)检测反应,结束后旋干溶剂,硅胶拌样,柱层析(PE/EA,2/1,v/v)得到化合物13d,白色固体,收率48%。1H NMR(500MHz,DMSO-d6)δ8.01(dd,J=8.6,0.9Hz,2H,ArH),7.90(t,J=7.5Hz,1H,ArH),7.75(t,J=8.1Hz,2H,ArH),4.61(s,1H,OH),4.46(t,J=6.3Hz,2H,CH2),3.53(q,J=5.4Hz,2H,CH2),1.90(p,J=12.3,6.0Hz,2H,CH2).
(g)将化合物Az-6(100mg,0.176mmol)溶于无水THF中,加入TBTU(68mg,0.212mmol)与DMAP(43mg,0.352mmol)待原料溶解后置换出体系内空气,搅拌0.5h后加入13d-h与12a(0.212mmol),氮气保护下反应6h,TLC(CH2Cl2/MeOH,20/1,v/v)检测反应,结束后旋干溶剂硅胶拌样,柱层析(CH2Cl2/MeOH,30/1,v/v),得到目标化合物2d,无色油状物,收率36%。1H NMR(CDCl3,500MHz)8.02-8.08(m,3H,ArH),7.75-7.76(m,2H,ArH),7.57-7.64(m,3H,ArH),7.17-7.18(d,J=8.2Hz 2H,ArH),7.05-7.07(d,J=8.2Hz,2H,ArH),6.78-6.89(m,6H,ArH),4.46-4.48(m,3H,-OCH2-,-CH2-),4.28-4.31(t,J=6Hz,2H,-CH2O-),4.11-4.18(m,5H,-OCH2-,-CH2-),3.59(s,2H,ArCH2-),3.31-3.34,3.42-3.45(dd,J=13.7Hz,2H,-NCH2-),2.82-2.85,3.07-3.09(dd,J=13.7Hz,2H,-CH2N-),2.22-2.26(m,2H,-CH2-),2.04(s,3H,NCH3),1.88-1.92(m,2H,-CH2-),1.77-1.82(m,2H,-CH2-).13C NMR(126MHz,CDCl3)δ171.72,163.77-163.67,161.78-161.69(dd,J=247.5,12Hz),159.98-159.89 158.02-157.93(dd,J=247.5,12Hz),158.82,158.36,158.11,151.02,144.66,138.11,135.66,133.86,130.25(s,2C),130.22(s,2C),129.88,129.75(s,2C),129.64-129.59,129.56-129.52(dd,J=9,6Hz),129.32,128.56(s,2C),127.89,125.98,114.66(s,2C),114.51(s,2C),111.63-111.60,111.46-111.44(dd,J=20.25,3Hz),110.46,104.45-104.24,104.23-104.03(dd,J=25.5Hz),72.09-72.05(d,1C),67.85,67.03,64.50-64.49(d,2C),62.71,60.51,56.54,56.50,43.65,40.48,31.97,29.74,29.34,28.28,27.85,22.73,14.16.HRMS(ESI-MS m/z)calculated for C42H44F2N6O10S[M+H]+863.2880,found863.2875ppm error:0.38.
实施例11
4-(5-(2-(4-(3-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基)(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)戊基)氧基)-3-(苯基磺酰基)-1,2,5-恶二氮-2-氧化物(2e)的制备
参照实施例10的合成方法。1H NMR(CDCl3,500MHz)8.11(s,1H,ArH),8.05-8.07(d,J=6.5Hz,2H,ArH),7.75-7.76(m,2H,ArH),7.57-7.64(m,3H,ArH),7.17-7.18(d,J=8.2Hz 2H,ArH),7.05-7.07(d,J=8.2Hz 2H,ArH),6.78-6.89(m,6H,ArH),4.44-4.50(dd,J=13.7Hz,-CH2-),4.40-4.43(t,J=6Hz,2H,-CH2O-),4.11-4.18(m,6H,-OCH2-),3.59(s,2H,ArCH2-),3.31-3.34,3.42-3.45(dd,J=13.7Hz,2H,-NCH2-),2.82-2.85,3.07-3.09(dd,J=13.7Hz,2H,-CH2N-),2.22-2.26(m,2H,-CH2-),2.04(s,3H,NCH3),1.84-1.92(m,2H,-CH2-),1.71-1.77(m,2H,-CH2-),1.49-1.56(m,2H,-CH2-).13C NMR(126MHz,CDCl3)δ171.99,163.76-163.67,161.78-161.69(dd,J=247.5,12Hz),159.98-59.89,158.02-157.93(dd,J=247.5,12Hz),159.03,158.36,158.06,151.02,144.66,138.19,135.63,130.33(s,2C),130.21(s,2C),129.88,129.69(s,2C),129.64-129.60,129.57-129.52(dd,J=8.75,6.25Hz),128.56,126.29,114.66(s,2C),114.50(s,2C),111.62-111.60,111.46-111.43(dd,J=20,3.75Hz),110.52,104.44-104.03(t,J=26.5Hz),72.10-72.05(d,1C),71.32,64.50(s,2C),64.37,62.71,61.14,56.54-56.50(d,1C),43.65,40.56,29.74,29.35,28.14,28.05,22.20.HRMS(ESI-MS m/z)calculated for C43H46F2N6O10S[M+H]+877.2964,found 863.3034ppm error:0.79.
实施例12
4-(2-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氨基)乙氧基)-3-(苯磺基尼龙基)-1,2,5-恶二唑2-氧化物(2f)的制备
参照实施例10的合成方法。1H NMR(CDCl3,500MHz)8.11(s,1H,ArH),8.05-8.07(d,J=6.5Hz,2H,ArH),7.75-7.76(m,2H,ArH),7.57-7.64(m,3H,ArH),7.17-7.18(d,J=8.2Hz,2H,ArH),7.05-7.07(d,J=8.2Hz,2H,ArH),6.78-6.89(m,6H,ArH),4.44-4.50(dd,J=13.7Hz,-CH2-),4.40-4.43(t,J=6Hz,2H,-CH2O-),4.11-4.18(m,4H,-OCH2-),4.12-4.14(t,J=13.7Hz,2H,-OCH2-),3.59(s,2H,ArCH2-),3.31-3.34,3.42-3.45(dd,J=13.7Hz,2H,-NCH2-),2.82-2.85,3.07-3.09(dd,J=13.7Hz,2H,-CH2N-),2.22-2.26(m,2H,-CH2-),2.04(s,3H,NCH3),1.84-1.92(m,2H,-CH2-),1.85-1.91(m,2H,-CH2-),1.66-1.71(m,2H,-CH2-),1.39-1.52(m,4H,-CH2-).13C NMR(126MHz,CDCl3)δ171.99,163.77-163.67,161.78-161.69(dd,J=247.5,12Hz),159.98-159.89,158.02-157.93(dd,J=247.5,12Hz),159.07,158.36,158.04,151.01,144.66,138.21,135.62,130.32(s,2C),130.21(s,2C),129.87,129.68(s,2C),129.64-129.59,129.57-129.52(dd,J=8.75,6.25Hz),128.57,126.36,114.65(s,2C),114.50(s,2C),111.62-111.60,111.46-111.43(dd,J=20,3.75Hz),110.52,104.44-104.24,104.23-104.03(dd,J=26.5Hz),72.09-72.05(d,1C),71.45,64.60,64.51,64.49,62.71,61.14,61.11,56.54-56.51(d,1C),43.64,40.59,29.74,29.35,28.49,28.36,25.46,25.28.HRMS(ESI-MS m/z)calculatedfor C44H48F2N6O10S[M+H]+891.3193,found 891.3183ppm error:1.27.
实施例13
4-(2-(2-(4-(3-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氨基)乙氧基)-3-(仅苯基磺)-1,2,5-氧杂二氮唑-2-氧化物(2g)的制备
参照实施例10的合成方法。1H NMR(CDCl3,500MHz)8.11(s,1H,ArH),8.05-8.07(d,J=6.5Hz,2H,ArH),7.75-7.76(m,2H,ArH),7.57-7.64(m,3H,ArH),7.17-7.18(d,J=8.2Hz,2H,ArH),7.05-7.07(d,J=8.2Hz,2H,ArH),6.78-6.89(m,6H,ArH),4.44-4.50(dd,J=13.7Hz,-CH2-),4.11-4.18(m,4H,-OCH2-),4.12-4.14(t,J=13.7Hz,2H,-OCH2-),3.67-3.68(m,3H,CONHCH2-),3.59(s,2H,ArCH2-),3.31-3.34,3.42-3.45(dd,J=13.7Hz,2H,-NCH2-),2.82-2.85,3.07-3.09(dd,J=13.7Hz,2H,-CH2N-),2.22-2.26(m,2H,-CH2-),2.04(s,3H,NCH3).13C NMR(126MHz,CDCl3)δ171.92,163.82(d,J=60.0Hz),159.97(d,J=50.0Hz),158.75,158.36,157.91(d,J=50.0Hz),151.03,144.69,137.91,135.72,130.55,130.23,129.74,129.61,128.61,126.53,115.18,114.54,111.63(d,J=70.0Hz),110.45,104.25(t,J=105.0Hz),72.11,70.08,64.48,62.71,61.05,56.52,43.63,42.87,38.40,29.30.HRMS(ESI-MS m/z)calculated for C40H41F2N7O9S[M+H]+834.2727,found834.2708ppm error:2.21.
实施例14
4-(4-(4-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)丁-2-炔基-1-基)氧基)-3-(PH-烯基磺酰基)-1,2,5-恶二唑2-氧化物(2h)的制备
参照实施例10的合成方法。1H NMR(500MHz,Chloroform-d)δ8.08(s,1H,ArH),8.06(dd,J=8.5,1.3Hz,2H,ArH),7.77-7.71(m,2H,ArH),7.64-7.53(m,3H,ArH),7.19(d,J=8.6Hz,2H,ArH),7.03(d,J=7.8Hz,2H,ArH),6.88(d,J=8.6Hz,2H,ArH),6.83-6.74(m,4H,ArH),5.08(t,J=1.8Hz,2H,OCH2),4.74(t,J=1.8Hz,2H,CH2),4.48-4.41(m,2H,CH2),4.14(q,J=6.0Hz,4H,CH2),3.61(s,2H,ArCH2),3.40(d,J=12.8Hz,1H,NCH2),3.30(d,J=12.6Hz,1H,NCH2),3.05(d,J=13.3Hz,1H,CH2N),2.79(d,J=13.0Hz,1H,CH2N),2.28-2.20(m,2H,CH2),2.01(s,3H,NCH3).13C NMR(126MHz,CDCl3)δ171.01,158.38,158.36,158.22,158.01,157.97,157.92,151.03,144.67,137.94,135.73,130.37,130.22,129.74,128.69,125.47,114.76,114.52,111.60,111.47,104.45,104.24,104.03,83.99,78.65,72.09,64.49,62.72,61.11,58.64,56.50,52.21,43.64,40.07,29.34.HRMS(ESI-MS m/z)calculated for C42H40F2N6O10S[M+H]+859.2567,found 859.2547ppm error:1.48.
实施例15
4-(2-(2-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)乙氧基)乙氧基)-3-(苯丙氨酸-尼龙磺酰基)-1,2,5-恶二唑2-氧化物(2i)的制备
参照实施例10的合成方法。1H NMR(CDCl3,500MHz)8.11(s,1H,ArH),8.05-8.07(d,J=6.5Hz,2H,ArH),7.75-7.76(m,2H,ArH),7.57-7.64(m,3H,ArH),7.17-7.18(d,J=8.2Hz,2H,ArH),7.05-7.07(d,J=8.2Hz,2H,ArH),6.78-6.89(m,6H,ArH),4.44-4.50(dd,J=13.7Hz,-CH2-),4.40-4.43(t,J=6Hz,2H,-CH2O-),4.54-4.57(t,J=8Hz,2H,-CH2O-),4.30-4.33(t,J=8Hz,2H,-CH2O-),4.11-4.18(m,4H,-OCH2-),3.87-3.90(t,J=8Hz,2H,-OCH2-),3.79-3.82(t,J=8Hz,2H,-CH2O-),3.59(s,2H,ArCH2-),3.31-3.34,3.42-3.45(dd,J=13.7Hz,2H,-NCH2-),2.82-2.85,3.07-3.09(dd,J=13.7Hz,2H,-CH2N-),2.22-2.26(m,2H,-CH2-),2.04(s,3H,NCH3).13C NMR(75MHz,CDCl3)δ171.89,164.39-164.23,161.09-160.92(dd,J=247.5,12Hz),160.60-160.44,157.33-157.18(dd,J=247.5,12Hz12Hz),158.91,158.29,158.01,150.96,144.63,138.03,135.65,133.83,130.35(s,2C),130.19(s,2C),129.79,129.67,129.64(s,2C),129.56-129.51,129.43-129.24(dd,J=8.75,6.25Hz),128.57(s,2C),127.92,126.00,114.56(s,2C),114.42(s,2C),111.65-111.61,111.38-111.34(dd,J=20,3.75Hz),104.55-103.86(t,J=26.5Hz),72.01-71.94(d,1C),70.54,69.37,69.12,68.56,68.32,65.86,64.39,63.81,62.64,61.03,60.98,56.49-56.43(d,1C),43.59,40.28,29.27,15.31.HRMS(ESI-MS m/z)calculated forC42H44F2N6NaO11S[M+Na]+901.2649,found 901.2613ppm error:-4.99.
实施例16
2-(二羟氧基)乙基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯(3a)的制备
(a)将2-溴乙醇18a(10mmol)溶于乙腈中,加入硝酸银(25mmol),在回流条件下反应过夜,TLC(PE/EA,10/1,v/v)检测,反应结束后待冷却至室温时加入过量饱和食盐水,有沉淀析出,硅藻土抽滤,滤液收集后使用EA(75mL)萃取三次,合并有机层后使用饱和食盐水洗涤,无水Na2SO4干燥,过滤,滤液经减压除去有机溶剂,残余物即为化合物19a,无色液体,收率92%。1H NMR(500MHz,Chloroform-d)δ4.59(t,J=4.6Hz,2H,CH2),3.96-3.90(m,2H,CH2).
(b)于5℃下,将发烟硝酸9.5mL(0.22mol,13.9g)缓慢滴入乙酸酐20ml中滴毕,于-5~0℃搅拌30min,形成硝酸-乙酸酐混合液。将乙醇胺6.0mL(0.10mol,6.1g)加入配有机械搅拌、温度计、恒压漏斗及无水CaCl,干燥管的三颈瓶中,冷却至
10℃左右滴入冰醋酸6.7mL(0.12mol);滴毕,搅拌15min,搅拌至固体溶解,转入恒压漏斗,于0~5℃下滴入硝酸-乙酸酐混合液中;滴毕,继续搅拌15min,于室温再搅拌4~5h,过滤,乙醚洗涤,干燥,异丙醇重结晶。活性炭脱色,得到白色片状晶体,收率87.1%。1HNMR(500MHz,DMSO-d6)δ7.98(s,2H,NH2),4.72(t,J=5.1Hz,2H,CH2),3.23(t,J=5.2Hz,2H,CH2).
(c)将化合物Az-6(100mg,0.176mmol)溶于无水THF中,加入TBTU(68mg,0.212mmol)与DMAP(43mg,0.352mmol)待原料溶解后置换出体系内空气,搅拌10min后加入19a(0.212mmol),氮气保护下反应6h,TLC(CH2Cl2/MeOH,20/1,v/v)检测反应,结束后旋干溶剂加入EA(30mL)后,超声使其全部溶解,用饱和食盐水萃取5次,收集有机层,无水Na2SO4干燥后过滤,滤液经减压除去有机溶剂,硅胶拌样,柱层析(DCM/MeOH,50/1,v/v),得到目标化合物3a,白色粘稠固体,收率34%。1H NMR(CDCl3,500MHz)2.05(s,3H,-NCH3),2.27-2.31(m,2H,-CH2-),2.83-2.86,3.08-3.11(dd,J=13.7Hz,2H,-CH2N-),3.33-3.35,3.44-3.46(dd,J=13.7Hz,2H,-NCH2-),3.62(s,2H,ArCH2-),4.17-4.20(m,4H,-OCH2-),4.39-4.41(t,J=5Hz,2H,-COOCH2-),4.45-4.52(dd,J=14.5Hz,2H,-CH2-),4.68-4.69(t,J=5Hz,2H,-CH2ONO2),6.79-6.87(m,4H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.58-7.63(m,1H,ArH),7.77(s,1H,ArH),8.12(s,1H,ArH).13C NMR(75MHz,CDCl3)δ171.39,164.42-164.26,161.12-160.96(dd,J=247.5,12Hz),160.64-160.48,157.30-157.21(dd,J=247.5,12Hz),158.35(s,2C),158.18(s,2C),150.88,144.58,130.24(s,2C),130.13(s,2C),129.85,129.63-129.55,129.51-129.43(dd,J=9,6Hz),126.46-126.30(d,J=12Hz),125.53,114.76(s,2C),114.52(s,2C),111.56-111.29(d,J=20.25Hz),104.48-103.78(t,J=26.25Hz),74.38,72.17-72.10(d,1C),,70.28,64.52,62.67,61.19,60.48,56.53-56.48(d,1C),43.59,40.04,32.29,29.31,0.95.HRMS(ESI-MS m/z)calculated for C32H35F2N5O8[M+H]+656.2548,Found656.2542PPM error:-0.91.
实施例17
3-(硝氧基)丙基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯(3b)的制备
参照实施例16的合成方法。1H NMR(CDCl3,500MHz)2.00(s,3H,-NCH3),2.02-2.04(m,2H,-CH2-),2.27-2.31(m,2H,-CH2-),2.83-2.86,3.08-3.11(dd,J=13.7Hz,2H,-CH2N-),3.33-3.35,3.44-3.46(dd,J=13.7Hz,2H,-NCH2-),3.62(s,2H,ArCH2-),4.17-4.20(m,4H,-OCH2-),4.13-4.15(t,J=5Hz,2H,-COOCH2-),4.43-4.46(m,4H,-CH2-,-CH2ONO2),6.79-6.87(m,4H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.58-7.63(m,1H,ArH),7.77(s,1H,ArH),8.12(s,1H,ArH).13C NMR(75MHz,CDCl3)δ171.52,164.40-164.24,161.10-160.94(dd,J=247.5,12Hz),160.64-160.49,157.37-157.21(dd,J=245.75,12Hz),158.34,158.12,156.14,150.84,144.57,130.19(s,2C),130.13(s,2C),129.86,129.64-129.56,129.52-129.44(dd,J=9,6Hz),126.48-126.30(d,J=12Hz),125.95,114.72(s,2C),114.52(s,2C),111.55-111.51,111.28-111.23(dd,J=20.25,3Hz),104.45-103.76(t,J=25.5Hz),72.21-72.14(d,1C),70.50,69.78,64.53,62.65,61.23,60.62,56.52-56.44(d,1C),53.39,43.59,40.33,29.31,26.40,0.95.HRMS(ESI-MS m/z)calculated forC33H37F2N5O8[M+H]+670.2670,Found 670.2683PPM error:1.94.
实施例18
4-(硝氧基)丁基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯(3c)的制备
参照实施例16的合成方法。1H NMR(CDCl3,500MHz)1.72-1.73(m,4H,-CH2-),2.00(s,3H,-NCH3),2.27-2.31(m,2H,-CH2-),2.83-2.86,3.08-3.11(dd,J=13.7Hz,2H,-CH2N-),3.33-3.35,3.44-3.46(dd,J=13.7Hz,2H,-NCH2-),3.62(s,2H,ArCH2-),4.11-4.15(m,6H,-OCH2-,-COOCH2-),4.41-4.47(m,4H,-CH2-,-CH2ONO2),6.79-6.87(m,4H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.58-7.63(m,1H,ArH),7.77(s,1H,ArH),8.12(s,1H,ArH).13C NMR(125MHz,CDCl3)δ171.78,163.75-163.65,161.76-161.67(dd,J=247.5,12Hz),159.97-159.87,158.00-157.91(dd,J=245.5,12Hz),158.35,158.10,150.94,144.62,130.24(s,2C),130.17(s,2C),129.86,129.61-129.57,129.54-129.49(dd,J=9,6Hz),126.50-126.47,126.40-126.37(dd,J=12.5,3.75Hz),126.11,114.68(s,2C),114.50(s,2C),111.57-111.55,111.41-111.38(dd,J=20.25,3Hz),104.39-103.98(t,J=25.5Hz),73.15,(s,1C),72.56,72.10-72.05(d,1C),64.50,63.78,62.68,61.81,61.14-61.10(d,1C),43.61,40.49,28.66,24.93,23.62.HRMS(ESI-MS m/z)calculated for C34H39F2N5O8[M+H]+684.2828,Found 684.2823PPM error:-0.73.
实施例19
5-(硝氧基)戊基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三偶氮-l-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯(3d)的制备方法
参照实施例16的合成方法。1H NMR:(CDCl3,500MHz)1.42-1.48(m,2H,-CH2-),1.66-1.70(m,2H,-CH2-),1.73-1.77(m,2H,-CH2-),2.05(s,3H,-NCH3),2.27-2.31(m,2H,-CH2-),2.83-2.86,3.08-3.11(dd,J=13.7Hz,2H,-CH2N-),3.33-3.35,3.44-3.46(dd,J=13.7Hz,2H,-NCH2-),3.62(s,2H,ArCH2-),4.11-4.14(t,J=6.5Hz,2H,-COOCH2-),4.17-4.19(m,4H,-OCH2-),4.42-4.45(t,J=6.5Hz,2H,-CH2ONO2),4.48-4.51(dd,J=13.7Hz,2H,-CH2-),6.79-6.87(m,4H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.58-7.63(m,1H,ArH),7.77(s,1H,ArH),8.12(s,1H,ArH).13CNMR(75MHz,CDCl3)δ171.76,164.42-164.26,161.32-161.13(dd,J=232.5,12Hz),160.72-160.50,158.36-158.06(dd,J=251.5,12Hz),157.37,157.27,150.92,144.58,130.22(s,2C),130.13(s,2C),129.89,129.64-129.44(dd,J=9,6Hz),126.53-126.30(dd,J=12.5,3.75Hz),114.67(s,2C),114.53(s,2C),111.57-111.30(d,J=20.25,3Hz),104.48-103.78(t,J=25.5Hz),73.19,72.94,72.18,64.17,64.55,62.69,62.22,61.23,43.60,40.53,29.34,28.07,26.39,22.20.HRMS(ESI-MS m/z)calculatedfor C35H41F2N5O8[M+H]+698.2984,Found 698.2981PPM error:-0.43.
实施例20
4-((硝氧基)甲基)苄基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯(3e)的合成方法
参照实施例16的合成方法。1H NMR(CDCl3,500MHz)2.05(s,3H,-NCH3),2.27-2.31(m,2H,-CH2-),2.83-2.86,3.08-3.11(dd,J=13.7Hz,2H,-CH2N-),3.33-3.35,3.44-3.46(dd,J=13.7Hz,2H,-NCH2-),3.62(s,2H,ArCH2-),4.17-4.19(m,4H,-OCH2-),4.48-4.51(dd,J=13.7Hz,2H,-CH2-),5.16(s,2H,-COOCH2-),5.45(s,2H,-CH2ONO2),6.79-6.87(m,4H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.36-7.37(d,J=8Hz,2H,ArH),7.40-7.41(d,J=8Hz,2H,ArH),7.58-7.63(m,1H,ArH),7.77(s,1H,ArH),8.12(s,1H,ArH).13C NMR(75MHz,CDCl3),δ,171.32,164.42-164.26,161.11-160.97(dd,J=247.5,12Hz),160.65-160.50,157.38-157.12(dd,J=247.5,12Hz),158.37,158.05,150.90,144.59,135.17,130.99,130.26(s,6C),130.16(s,2C),129.81,129.66-129.46(dd,J=9,6Hz),128.94,126.49-126.24(dd,J=12.5,3.75Hz),125.87,114.70(s,2C),114.55(s,2C),111.56-111.30(dd,J=20.25,3Hz),104.49-103.79(t,J=25.5Hz),72.19,64.55,63.91,62.68,61.22,56.48,51.84,43.61,40.34,29.33,0.99.HRMS(ESI-MS m/z)calculated for C38H39F2N5O8[M+H]+732.2834,Found 732.2839PPM error:0.68
实施例21
2-((硝氧基)甲基)苄基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯(3f)的制备方法
参照实施例16的合成方法:1H NMR(CDCl3,500MHz)2.05(s,3H,-NCH3),2.27-2.31(m,2H,-CH2-),2.83-2.86,3.08-3.11(dd,J=13.7Hz,2H,-CH2N-),3.33-3.35,3.44-3.46(dd,J=13.7Hz,2H,-NCH2-),3.62(s,2H,ArCH2-),4.17-4.19(m,4H,-OCH2-),4.48-4.51(dd,J=13.7Hz,2H,-CH2-),5.20(s,2H,-COOCH2-),5.44(s,2H,-CH2ONO2),6.79-6.87(m,4H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.36-7.38(m,4H,ArH),7.58-7.63(m,1H,ArH),7.77(s,1H,ArH),8.12(s,1H,ArH).13C NMR(75MHz,CDCl3)δ171.52,164.42-164.26,161.11-160.96(dd,J=247.5,12Hz),160.65-160.49,158.11-158.04(dd,J=245.5,12Hz),158.34,157.16,150.85,144.57,137.41,132.22,130.29(s,6C),130.25(s,2C),130.14,129.90,129.64-129.56,129.52-129.44(dd,J=9,6Hz),129.17,129.13,128.36-128.25,127.16-127.06(dd,J=12.5,3.75Hz),126.22,120.02,114.70(s,2C),114.54(s,2C),111.58-111.29(dd,J=20.25,3Hz),104.49-103.79(t,J=25.5Hz),74.61,61.19,74.31,72.19,56.52,65.83,64.55,62.68,43.62,40.25,29.33.HRMS(ESI-MS m/z)calculated for C38H39F2N5O8[M+H]+732.2835,Found 732.2839PPM error:0.55.
实施例22
(3S,3aR,6R,6aS)-6-(硝氧基)六氢呋喃[3,2-b]呋喃-3-基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)氨基)甲基-基苯氧基)丙氧基)氢烯基)乙酸酯(3g)的制备
参照实施例16的合成方法。1H NMR(CDCl3,500MHz)8.12(s,1H,ArH),7.77(s,1H,ArH),7.58-7.63(m,1H,ArH),7.20-7.22(d,J=8.5Hz,2H,ArH),7.06-7.08(d,J=8.5Hz,2H,ArH),6.90-6.92(d,J=8.5Hz,2H,ArH),6.79-6.87(m,4H,ArH),4.91-4.93(m,1H,ISMN),5.31-5.32(m,1H,ISMN),4.39-4.46(m,3H,-CH2-,ISMN),4.12-4.14(m,4H,-OCH2-),3.95-4.00(m,3H,ISMN),3.91-3.92(m,1H,ISMN),3.84-3.88(m,1H,ISMN),3.54(s,2H,ArCH2-),3.28-3.30,3.39-3.41(dd,J=13.7Hz,2H,-NCH2-),2.78-2.81,3.03-3.05(dd,J=13.7Hz,2H,-CH2N-),2.22-2.26(m,2H,-CH2-),2.05(s,3H,-NCH3).13C NMR(75MHz,CDCl3)δ170.86,164.40-164.24,161.10-160.94(dd,J=247.5,12Hz),160.61-160.45,157.34-157.18(dd,J=247.5,12Hz),158.29,158.14,150.84,144.59,130.21(s,2C),130.17(s,2C),129.83,129.61-129.54,129.49-129.42(dd,J=9,6Hz),126.49-126.45,126.32-126.28(dd,J=12.5,3.75Hz),125.45,114.69(s,2C),114.44(s,2C),111.63-111.35(dd,J=20.25,3Hz),104.54-103.84(t,J=25.5Hz),89.02,86.54,81.60-81.50,81.27-81.14(dd,1C,J=24.75,7.5Hz),77.62,75.88,75.41,73.51,72.02-71.95(d,1C),,69.22,69.07,64.41(s,2C),62.63,61.02,60.38,56.51-56.44(d,1C),43.60,40.20,29.67,29.27,21.02,1.01.HRMS(ESI-MS m/z)calculated for C38H39F2N5O8[M+H]+740.2726,Found 740.2737PPM error:2.32.
实施例23
2-(2-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰基)-L2-氮杂基)乙基硝酸酯(3h)
参照实施例16的合成方法。1H NMR(300MHz,CDCl3)δ8.08(s,1H,ArH),7.72(s,1H,ArH),7.56(td,J=9.2,6.6Hz,1H,ArH),7.14(d,J=8.6Hz,2H,ArH),7.04(d,J=8.0Hz,2H,ArH),6.90(d,J=8.6Hz,2H,ArH),6.85-6.73(m,4H,ArH),4.50(t,J=5.1Hz,2H,CH2ONO2),4.43(s,2H,CH2),4.20-4.10(m,4H,OCH2),3.54(t,J=6.0Hz,2H,COOCH2),3.52(s,2H,ArCH2),3.35(dd,J=36.1,10.7Hz,2H,CH2),2.92(dd,J=78.0,15.0Hz,2H,CH2),2.27(p,J=6.1Hz,2H,CH2),2.01(s,3H,CH3),1.65(s,1H,NH).13C NMR(126MHz,CDCl3)δ171.94,163.86-163.76,161.87-161.76(dd,J=250.7,12.6Hz),159.94-159.85,157.98-157.89(dd,J=247.0,11.3Hz),158.38,151.08,144.73,130.55,130.10,129.67,129.62-129.55(t,J=6.3Hz),126.40,115.21,114.69,114.62,111.73-111.59(d,J=17.6Hz),104.58-104.50(t,J=26.5Hz),72.15-72.11(d,J=5.0Hz),71.55,67.73,64.51,64.43,62.65,60.85,56.41,43.61,42.73,37.17,33.88,29.31,1.06.HRMS(ESI-MS m/z)calculated forC32H37F2N6O7[M+H]+655.2686,Found 655.2670PPM error:2.37.
实施例24
体外一氧化氮释放实验
实验方法:称量化合物3a或3h,溶解于40μL DMSO溶液中,配置成浓度为50mM的DMSO母液,吸收DMSO母液40μL,加入大鼠血浆3960μL。化合物的终浓度为500μM。将溶液于37℃条件下孵育,分别于0h、1h、2h、4h、8h、12h、24h吸收200μL溶液,加入200μL甲醇,离心,吸收300μL上清,加入100μLGriess试剂,离心,吸收上清,过滤。取滤液200μL加入96孔板,在540nm波长处测定吸光度。将NaNO2称重并配置成不同浓度梯度的大鼠血浆溶液。按上述操作方法测定不同浓度亚硝酸钠溶液的吸光度,并绘制浓度-吸光度标准曲线。将化合物的吸光度数据代入标准曲线,计算不同时间点的NO释放率。对于化合物1a和2i,称取化合物溶解在1×PBS溶液中,最终浓度为100μM。加入不同浓度的谷胱甘肽,混合均匀,37℃孵育,在给定时间吸收100μL溶液,在96孔板中加入100μL Griess试剂,在540nm波长处测量吸光度。
实验结果:结果如附图2所示,在GSH条件下,图2a和图2b中化合物1a和2i能很好的释放NO并且随着GSH浓度升高释放率增加,孵育24小时后,释放率分别达到了60%和14%以上。图2c中,化合物3a和3h在大鼠血浆中37℃条件下孵育24小时,均具有显著NO释放水平。
实施例25
体外抗真菌活性分析
实验方法:MIC80的测定采用美国临床实验室标准化协会(Clinical andLaboratory Standards Institute,CLSI)M27-A3和M38-A2文件所推荐的微量液基稀释方法进行测定,并通过培养物的光密度值(OD)来评价真菌的生长情况,以此考察化合物对真菌的体外抑制活性。用RPMI 1640培养液配制1×103cells/mL的菌悬液,然后将配制好的菌悬液涡旋混匀后加入96孔细胞培养板中,每孔加入100μL,化合物从浓度64μg/mL开始依次倍半稀释,使其终浓度为64~0.125μg/mL。以不加任何药物作用的菌悬液为阴性对照,以RPMI 1640培养液为空白对照,将96孔细胞培养板置于30℃恒温培养箱中静置培养(隐球菌72h,其它菌株48h),培养完成后使用酶标仪测定每孔真菌在630nm处的光密度值(OD630),平行测定三次。取抑制率≥80%时所对应的最小浓度即为该化合物的最低抑菌浓度值(MIC80)。
实验结果:如附图3a和表1所示。化合物2a-2i对白色念珠菌、光滑念珠菌和新型隐球菌具有中等抑菌活性(MIC80=1~16μg/mL)。化合物3a-3h具有良好的抑菌活性(MIC80=0.125~4μg/mL)。
表1.NO供体型化合物和FLC的体外抗真菌活性(MIC80,μg/mL)
aAbbreviations:C.alb.,Candida albicans;C.gla.,Candida glabrata;C.neo.,Cryptococcus neoformans;FLC,fluconazole.
实施例26
时间-生长曲线试验
实验方法:取已在显微镜下计数好的真菌细胞原液于96孔细胞培养板中依次倍半稀释,采用酶标仪测量各孔的OD630,根据不同OD630值(x)及其相对应的菌浓度(y)值作图得到标准曲线。用RPMI 1640培养液配制菌悬液使其浓度为1×105cells/mL。用移液枪吸取5mL配制好的菌悬液加入15mL离心管中同时加入不同浓度的化合物,用FLC作为阳性对照且以DMSO作为空白对照,然后将15mL离心管放入30℃气浴恒温振荡培养箱中以200rpm的转速振荡培养。在设计的培养时间点(0h、3h、6h、9h、12h、24h、48h、72h)从离心管中吸取100μL真菌细胞培养液于96孔细胞培养板(三复孔),采用酶标仪测量各孔的OD630值,利用标准曲线拟合的方程计算各组的菌浓度并采用Graphpad Prism 8作图。
实验结果:结果如附图3b所示。化合物3a、3b和3e抑制新生隐球菌的生长,浓度低至2μg/mL时也能完全抑制细胞生长,明显优于氟康唑FLC。
实施例27
时间-杀菌曲线实验
实验方法:将呈指数增长的新生隐球菌H99细胞用PBS洗涤,用RPMI 1640培养基重悬至1.0×105个/mL。将不同浓度的FLC和化合物3a、3e加入到新型隐球菌H99菌株悬浮液中。添加FLC和所选化合物均不作为对照组。加入不同浓度的化合物和FLC,在30℃的摇箱(200rpm)中孵育。在设定时间(0、6、12、24、48、72h),将悬浮液包被在SDA板上,30℃孵育72h,计数板上的单克隆真菌菌落数,完成3次独立实验。
实验结果:如附图3c所示。在4μg/mL浓度下,FLC无杀真菌作用,与文献报道一致。而化合物3a和3e对新型隐球菌有显著的杀真菌作用,浓度为8μg/mL可以完全杀死新生隐球菌。
实施例28
平皿扩散实验
实验方法:将约1×105个细胞置于含有不同浓度FLC或化合物3a、3e的SDA培养基上。35℃孵育72h后拍照,直观反映新生隐球菌H99菌株的抗真菌活性。
实验结果:如附图3d所示,化合物3a和3e在菌落培养72小时后,浓度为0.125-1μg/mL时,完全抑制新生隐球菌的生长。而在相同浓度下,FLC对新生隐球菌细胞的生长几乎没有抑制作用。
实施例29
体外生物膜形成抑制试验
实验方法:将菌株的悬液(RPMI 1640培养基中为1.0×106个细胞/mL)加入96孔板(每孔100μL),37℃孵育3小时使其粘附。用PBS洗涤贴壁细胞3次,去除上层培养基和浮游植物细胞。同时,在新鲜RPMI 1640培养基中加入不同浓度的FLC和化合物,37℃下再孵育24h。采用XTT还原法计算半定量测定生物膜的形成。利用Microplate Reader测定490nm处的光密度(OD490),并完成3个独立实验。
实验结果:如附图4所示。化合物3a、3b和3e在0.25μg/mL浓度下显著抑制新生隐球菌生物膜的形成(P<0.0001),抑制率达到50%以上,抑制活性显著优于FLC(P<0.01)(图4a)。为了研究NO释放对生物膜形成的影响,在处理组添加NO清除剂2-苯基-4,4,5,5-四甲基咪唑啉-1-氧基3-氧化物(PTIO)。结果表明,添加PTIO后,在0.25μg/mL浓度下,化合物3a对生物膜形成的抑制作用显著减弱(P<0.01,图4b)。同时,我们还研究了化合物3a和3e预先形成的成熟生物膜的破坏情况(图4c和图4d)。FLC对已成型的成熟生物膜的破坏作用较弱,而化合物3a和3e在浓度为0.125μg/mL时可破坏约25%的预成型成熟生物膜,高浓度(64μg/mL)时可破坏约50%的预成型成熟生物膜。此外,添加NO清除剂PTIO后,化合物3a和3e的破坏作用可被明显减弱,这结果表明化合物3a和3e对新生隐球菌H99菌株生物被膜的形成具有显著的抑制作用,且该抑制作用依赖于NO的释放。
实施例30
一氧化氮释放实验
实验方法:收集指数生长的细胞,用PBS洗涤三次,并在1.0×107个细胞/mL的RPMI1640培养基中重悬。然后,逐渐向悬浮液中加入不同浓度的化合物。37℃,5%CO2孵育12h后,离心收集新生隐球菌H99细胞,加入适量裂解液。随后,用DAF-FM DA(1mL,10μM)在37℃下黑暗重悬40min。用PBS冲洗细胞3次,然后用激光扫描共聚焦显微镜(Leica SP5,Germany)分析,激发波长为495nm,发射波长为515nm。
实验结果:3a和3e处理组真菌细胞大部分呈绿色荧光,表明NO释放。同时,经3a和3e处理后,大多数真菌细胞也可见红色荧光,表明细胞死亡。合并后,在许多细胞中观察到红色和绿色荧光重叠(图5)。这表明3a和3e都是通过穿透真菌细胞,释放NO,破坏真菌细胞膜的完整性,从而导致真菌细胞死亡来发挥抗真菌作用的。
实施例31
真菌麦角甾醇含量实验
实验方法:甾醇成分分析采用气相色谱-质谱法,通过利用气相色谱-质谱图各峰的分子片段与国家标准技术研究院参考数据库中相应的甾醇化合物匹配,实现甾醇的鉴定。样品处理步骤如下:将菌株(50μL)加入到50ml YEPD培养基中,制备浓度为1.0×106个细胞/mL的悬液。同时,在培养液中加入化合物和FLC对新生隐球菌H99细胞进行处理。35℃孵育24h后,离心收获细胞,用10mL20%NaOH(w/v)在80℃90%乙醇中皂化3h,然后在皂化混合物中加入10mL正己烷提取固醇。然后,减压蒸发溶剂,用400μL环己烷作为样品溶解残渣。最后采用GC-MS分析各组的甾醇成分。
实验结果:甾醇含量结果(图6a和图6b)显示,空白对照组的麦角甾醇含量为58.33%;经FLC(4μg/mL)作用后麦角甾醇含量降至19.21%;硝酸酯唑类衍生物3a和3e在0.125μg/mL时分别能将麦角甾醇含量降低至23.68%和10.66%。空白对照组的羊毛甾醇含量为6.24%,经FLC(4μg/mL)作用后羊毛甾醇含量升高至53.85%,经0.125μg/mL的3a和3e作用后,羊毛甾醇含量分别升高至44.52%和50.94%。甾醇含量分析结果表明化合物3a、3e通过破坏真菌细胞膜上的麦角甾醇的生成并产生大部分的羊毛甾醇而发挥抗真菌活性。
实施例32
透射电镜实验观察荚膜形成
实验方法:用YEPD培养液配制菌悬液使其浓度为1×106cells/mL,取15mL离心管若干,各加入10mL菌悬液并加入不同浓度化合物作用,以不加药组作为空白对照组。然后置于35℃恒温振荡培养箱中,以220rpm的转速振荡培养8h。培养完成后,离心,收集真菌细胞,使用PBS缓冲溶液洗涤3次,加入1mL电镜固定液,反复吹打、涡旋、混匀,使其自然沉降,于4℃固定过夜。采用透射电子显微镜对真菌细胞形态进行观察并拍照。
实验结果:结果如图6c所示,正常新生隐球菌H99细胞的密度均匀,轮廓清晰,具有完整的细胞膜、细胞壁以及致密的荚膜。经4μg/mL的FLC作用后,新生隐球菌H99细胞无明显损伤。经1μg/mL的化合物3a处理后,新生隐球菌H99的细胞膜完全破裂,细胞内容物渗出。结果表明化合物3a能够破坏新生隐球菌H99细胞膜的完整性。
实施例33
体内抗新生隐球菌活性实验
实验方法:如图7a所示,选用ICR雌性小鼠(体重:18-22g),通过尾静脉注射接种真菌进行造模(接种菌量:0.2mL/只,即2×105cells/只)。小鼠接种真菌感染24h后,采用滴鼻给药方式,在左右鼻孔黏膜处各滴加10μL化合物,连续给药五天,记录小鼠生存情况。于第六天处死小鼠,解剖取出小鼠的脑或肺等组织(需称重并记录),装入已编号的1.5mL离心管中,每管加入1mL生理盐水和2粒钢珠,然后使用组织研磨仪研磨均匀,于4℃保存。将脑或肺部组织液稀释适当倍数后涂板于SDA培养基上,将培养皿置于30℃恒温培养箱中静置培养72h。取出SDA培养皿进行单菌落计数,计算各组小鼠的脑或肺部组织荷菌量。借助作图分析软件Graphpad prism 8对各组之间的统计学差异进行ANOVA和post hoc(Bonferroni andStudent-Newman-Keuls')分析。同时对小鼠脑组织进行病理切片实验,将剖取的组织经4%多聚甲醛溶液固定、脱水、石蜡包埋、制片、PAS和H&E(hematoxylin-eosin)染色,在光学显微镜下观察并拍照记录。
实验结果:结果如图7b和图7c所示,给予化合物3a和3h(分别设置两个剂量:1mg/kg和10mg/kg)治疗后,小鼠脑组织荷菌量与对照组相比明显下降,且有统计学差异,P<0.001;当给药剂量为1mg/kg时,给予化合物3h组对小鼠脑组织荷菌量的抑制作用优于FLC给药组并具有显著性差异(P<0.05)。以上结果说明NO供体型唑类衍生物对小鼠脑组织荷菌量具有较好的抑制作用,且化合物3h体内活性显著优于3a。荷菌量实验结果表明化合物3a、3h具有优秀的体内抗新生隐球菌活性。
PAS(periodic acid-Schiff)染色结果显示,对照组小鼠脑组织出现不规则空腔和大量真菌细胞定植,提示大量新生隐球菌侵入脑组织。相比之下,经化合物3a和3h处理后可有效减少新生隐球菌侵入大脑的数量,进一步证明NO供体型衍生物具有优秀的体内抗真菌作用(图7d)。H&E(hematoxylin-eosin)染色结果显示,对照组小鼠脑组织有典型的病理改变,包括血管充血、神经元坏死、脑内不规则空洞。此外,还观察到多处神经元排列不规则,胶质细胞数量增加(图7e)。给予化合物3a和3h处理后,小鼠脑组织病理损伤明显减轻。上述研究结果表明NO供体型硝酸酯唑类衍生物具有较好的体内抗新生隐球菌效果,从而为隐球菌性脑膜炎的治疗提供了新的方向。
Claims (10)
1.一种NO供体型抗真菌化合物,其特征在于,所述化合物为通式V所示:
其中,A为
当A为其中/>为4-羟基哌啶基、二乙胺基、D-脯氨醇基、N-甲基哌嗪基、四氢吡咯基或N-甲基乙醇胺基;
当A为其中n=2,3,5;
当A为 X为脂肪烃或芳香烃。
2.根据权利要求1所述的化合物,其特征在于,当A为X为/>
3.根据权利要求1所述的化合物,其特征在于,当A为X为/>
4.根据权利要求1所述的化合物,其特征在于,所述化合物具体如下:
1a:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(4-羟基哌啶-1-基)二氮杂环形-1-鎓-1,2-二氢酸酯;
1b:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)胺-邻)-2,4-二硝基苯氧基-1-(S)-2-羟甲基吡咯烷-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯;
1c:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(吡咯烷-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯;
1d:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)氨基-邻)-2,4-二硝基苯;
1e:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基-O)-2,4-二硝基苯氧基-1-(4-甲基哌嗪-1-基)二氮杂环己烷-1-鎓-1,2-二氢酸酯;
1f:5-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)胺邻)-2,4-二硝基苯氧基-3-(2-羟乙基)-3-甲基二氮杂环戊烯-1-鎓-1,2-二氢酸酯;
2a:4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苄基)氨基)乙氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物;
2b:4-(3-(4-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苄基)氨基丙氧基)-3-苯磺酰基-1,2,5-恶二唑2-氧化物;
2c:4-((5-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基氨基)甲基)苄基)氨基)戊基)氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物;
2d:4-(4-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基)(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)丁氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物;
2e:4-(5-(2-(4-(3-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基)(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)戊基)氧基)-3-(苯基磺酰基)-1,2,5-恶二氮-2-氧化物;
2f:4-(2-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氨基)乙氧基)-3-(苯磺基尼龙基)-1,2,5-恶二唑2-氧化物;
2g:4-(2-(2-(4-(3-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基氨基)甲基)苯氧基)丙氧基)苯基)乙酰氨基)乙氧基)-3-(仅苯基磺)-1,2,5-氧杂二氮唑-2-氧化物;
2h:4-(4-(4-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙酰基(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)丁-2-炔基-1-基)氧基)-3-(PH-烯基磺酰基)-1,2,5-恶二唑2-氧化物;
2i:4-(2-(2-(2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰氧基)乙氧基)乙氧基)-3-(苯丙氨酸-尼龙磺酰基)-1,2,5-恶二唑2-氧化物;
3a:2-(二羟氧基)乙基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3b:3-(硝氧基)丙基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3c:4-(硝氧基)丁基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3d:5-(硝氧基)戊基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三偶氮-l-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3e:4-((硝氧基)甲基)苄基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3f:2-((硝氧基)甲基)苄基-2-(4-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酸酯;
3g:(3S,3aR,6R,6aS)-6-(硝氧基)六氢呋喃[3,2-b]呋喃-3-基-2-(4-((2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基(甲基)氨基)甲基-基苯氧基)丙氧基)氢烯基)乙酸酯;
3h:2-(2-(2-(2,4-二氟苯基)-2-羟基-3-(1H-1,2,4-三唑-1-基)丙基)(甲基)氨基)甲基)苯氧基)丙氧基)苯基)乙酰基)-L2-氮杂基)乙基硝酸酯。
5.一种权利要求1所述化合物的制法,其特征在于,具体如下:以仲胺类化合物4a-f为原料,高压下与NO气体反应得到偶氮鎓二醇钠盐类化合物5a-f,在过弱碱的作用下与2,4-二硝基氟苯发生亲核取代反应生成化合物6a-f;同时,取三甲基碘化亚砜在强碱环境加热生成碳负离子中间体C-H3,再与原料Az-1发生环氧化反应得到Az-2,Az-2在碱性条件下被胺类化合物开环得到Az-3,最后化合物6a-f和Az-3反应制得目标化合物;
6.一种权利要求1所述化合物的制法,其特征在于,具体如下:以苯硫酚钠7为原料,与氯乙酸反应得到苯硫基乙酸8,然后8氧化得到化合物9,除去溶剂后在高温下与发烟硝酸反应得到呋咱氮氧化物10,呋咱氮氧化物在碱性条件下与Boc保护的氨基醇反应得到11a-c,脱保护后即得到中间体12a-c;同时,中间体Az-3与4-溴甲基苯甲醛发生亲核加成反应得到Az-4;最后,12a-c和Az-4通过弱还原剂腈基硼氢化钠的还原胺化得到目标化合物;
7.一种权利要求1所述化合物的制法,其特征在于,具体如下:以对羟基苯乙酸甲酯14为原料与1,3-二溴丙烷在碱性条件下发生亲核加成得到15,15进一步与对羟基苯甲醛反应得到16,纯化后与甲胺盐酸盐在无水条件、三乙胺的催化下得到亚胺中间体,直接还原得到化合物17;将Az-2溶于乙腈中并加入过量三乙胺游离,后加入17在碱性环境下开环得到化合物Az-5,脱去乙酯部分即得到中间体Az-6;同时,由10与二醇在碱性环境下发生亲核取代得到13d-h;最后,Az-6和13d-h在催化剂的作用下缩合得到目标化合物;
8.一种权利要求1所述化合物的制法,其特征在于,具体如下:以端基溴醇为原料,在高温避光条件下与硝酸银反应生成羟基硝酸酯类化合物,后与Az-6一步酯缩合得到目标化合物;
9.一种药物组合物,由权利要求1所述的通式V的化合物或其医学上可接受的盐与药学上可接受的载体或辅料的药物组成。
10.一种权利要求1所述的化合物在制备抗真菌的药物中的应用。
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