CN113214161A - 一种具有抗拉沙病毒活性的化合物及其制备方法和应用 - Google Patents
一种具有抗拉沙病毒活性的化合物及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种具有抗拉沙病毒活性的化合物及其制备方法和应用,涉及抗病毒治疗药物技术领域。所述化合物为2,6‑二甲氧基‑4‑((1‑(4‑甲氧基苯基)‑1h‑苯并[d]咪唑‑5‑基氨基甲基)苯酚,是以2,4‑二硝基氟苯、对甲氧基苯胺为原料,经过碳酸铯催化反应后,再经过保险粉还原、甲酸反应,最终和丁香醛反应制得。本发明首次公开了该化合物及其在抗拉沙病毒药物中的应用,且其合成方法工艺简单,操作简单,质量可控,收率高,工艺平行性及重现性好,适合大规模化生产。
Description
技术领域
本发明涉及抗病毒治疗药物技术领域,尤其涉及一种具有抗拉沙病毒活性的化合物及其制备方法和应用。
背景
沙粒病毒属于RNA病毒,是出血热病毒中的一种。这种病毒颗粒在电镜超薄切片中有砂粒样结构,而因此得名。沙粒病毒分为旧世界沙粒病毒和新世界沙粒病毒,其中旧世界沙粒病毒主要包括淋巴细胞脉络丛脑膜炎病毒(Lymphocytic choriomeningitis virus,LCMV),拉沙病毒(Lassa virus,LASV),Lujo病毒(Lujo virus)等;新世界沙粒病毒主要包括呼宁病毒(Junin virus,JUNV),马秋博病毒(Machupo virus,MACV),瓜纳瑞托病毒(Guanarito virus,GTOV),白水阿罗约病毒(White Water Arroyo virus,WWAV)以及Sabia病毒(SABV)塔卡里伯病毒(Tacaribe virus)等。
拉沙病毒、呼宁病毒、马秋博病毒以及瓜纳瑞托病毒感染后主要引起人类出血热。美国疾病控制和预防中心(CDC)对可进行生物恐怖袭击的潜在致病微生物分类,认定拉沙病毒、呼宁病毒、马秋博病毒、瓜纳瑞托病毒为A级,即可能严重影响到公众的安全和健康。该类病毒传播快,病死率高,成为重要的潜在战剂病毒,极有可能成为生物武器。美国国家传染病研究所(NIAID)评定此类病毒为最具危险性的潜在生物武器之一,并要求尽快寻找有效的抗病毒治疗方法。此外,美国生物防御工作小组(Working Group on CivilianBiodefense)描述了包括拉沙病毒在内的几种出血热病毒具有被用作生物武器的潜在危险,并建议研究和开发新的抗沙粒病毒药物。
沙粒病毒基因组由两个单链RNA片段构成,各自编码反取向的两个基因。其中较大的片段L RNA(7.2kb)编码L和Z蛋白。L蛋白是RNA依赖性RNA聚合酶(RdRp),Z蛋白是小型锌结合环指蛋白,其参与病毒出芽。S RNA(3.4kb)编码核蛋白(NP)和包膜糖蛋白前体(GPC)。包膜糖蛋白被嵌入包围病毒核壳的脂双分子层。沙粒病毒糖蛋白(GP)功能是将病毒附着于特定的寄主细胞受体以及介导病毒膜与宿主膜的融合,从而在靶细胞内部淀积病毒基因组。包膜蛋白在整个内质网的膜内的共翻译转运水解进一步将加工成N-末端亚单位(GP1)其含有结合决定簇的受体,和C-末端跨膜亚单位(GP2),其能够经历与膜融合有关的显著的构型重排。这两个亚单位仍然彼此结合并被装配成该杂二聚体的丫聚体复合物,GP1和GP2形成异源二聚体,通过与旧世界病毒LASV和LCMV的α-营养不良聚糖基因相互作用或新世界病毒的的的转铁蛋白1受体促使其进入宿主细胞。
拉沙病毒是流行程度最广、危害性最大的沙粒病毒,主要流行于西非,每年感染者达10万~30万人,死亡约5000人。新世界型病毒中致出血热的几种沙粒病毒主要流行于美洲,死亡率约15%~30%。因此,仍然迫切需要开发安全和有效的产品以抵御可能的生物攻击以及拉沙病毒得爆发感染。
发明内容
针对上述问题,本发明的目的在于公开一种具有抗拉沙病毒活性的化合物及其制备方法和应用,对沙粒病毒感染,尤其是拉沙病毒感染及其相关疾病具有治疗效果,可以用于制备抗拉沙病毒的药物,且其合成方法,工艺简单,操作简单,质量可控,收率高,工艺平行性及重现性好,适合大规模化生产。
具体的,本发明的一种具有抗拉沙病毒活性的化合物,所述化合物的结构式如式(I)所示:
或它在药学上可接受的盐。
本发明还公开了一种药物组合物,包含有效剂量的式(I)表示的化合物或其药学可接受的盐和药学可接受的载体或赋形剂
本发明还公开了上述化合物或其药学上可接受的盐或药物组合物在治疗或预防哺乳动物的与沙粒病毒相关的疾病或疾病状态的药物中的应用,其中抗沙粒病毒被认为是有益的。
进一步,所述哺乳动物为人。
进一步,所述沙粒病毒为拉沙病毒、呼宁病毒、马丘波病毒、瓜纳瑞托病毒、萨比亚病毒、白水阿罗约病毒、查帕雷病毒、LCMV、LCMV样病毒中任意一种。优选的为拉沙病毒。
此外,本发明的另一个目的是提供上述具有抗拉沙病毒活性的化合物的制备方法,所述制备方法是以2,4-二硝基氟苯和对甲氧基苯胺为原料,在催化剂作用下反应得到中间体Ⅰ,将中间体Ⅰ经保险粉还原后得到中间体Ⅱ,再与甲酸反应后得到中间体Ⅲ,最后以中间体Ⅲ和丁香醛为原料,在三乙酰氧基硼氢化钠作用下反应得到产品2,6-二甲氧基-4-((1-(4-甲氧基苯基)-1h-苯并[d]咪唑-5-基氨基甲基)苯酚。
进一步,所述中间体Ⅰ和保险粉的质量比为1:6-1:9.1。
进一步,所述制备方法具体包括以下步骤:
所述制备方法具体包括以下步骤:
S1:将2,4-二硝基氟苯与对甲氧基苯胺加入反应釜中,再加入四氢呋喃,搅拌溶解后,加入催化剂碳酸铯,加热至48℃,保温反应10-12h,反应完成后,反应液过滤,滤液经减压蒸干后,得到第一残留物,将第一残留物加入乙酸乙酯搅拌至完全溶解后,加入饱和碳酸氢钠洗涤两次,有机层用无水硫酸镁干燥过夜,过滤、滤液减压蒸干,得到第二残留物,第二残留物加入石油醚/乙酸乙酯混合溶液进行重结晶,得到中间体Ⅰ;
S2:将中间体Ⅰ加入无水乙醇中,搅拌,加热至回流使其溶解,滴加保险粉水溶液,滴加完成后,持续保温至反应完成后,反应液继续搅拌20min,静置,冷却至室温过滤,滤液减压浓缩,得到第三残留物,第三残留物中加入无水乙醇,加热搅拌20min,过滤,滤液减压浓缩,得到中间体Ⅱ;
S3:将中间体Ⅱ加入4N盐酸搅拌溶解后,加入无水甲酸加热至回流,保温反应3-5h后,反应完成,减压蒸干溶剂,得到第四残留物加水溶解,调节pH至13,用二氯甲烷萃取后,收集有机层,有机层用无水硫酸镁干燥过夜,过滤后减压蒸干溶剂,得到第五残留物用无水乙醇溶解后,加入活性炭脱色,加热回流20min,趁热过滤,滤液浓缩至1/2体积,于0-4℃温度下冷却结晶,过滤、干燥得到中间体Ⅲ;
S4:将中间体Ⅲ、丁香醛加入1,2-二氯乙烷中,搅拌溶解后,加入醋酸,加热至60℃,搅拌3-5h,分批加入三乙酰氧基硼氢化钠,于60℃反应5-7h,反应液用饱和碳酸氢钠洗涤,有机层干燥,过滤,减压蒸干,得到第六残留物用通过硅胶快速柱色谱分离,收集目标组分,减压蒸馏除去溶剂,得到第七残留物加入乙酸乙酯/环己烷混合液,放置于4℃处冷却析晶,过滤,干燥,得到目的物,即2,6-二甲氧基-4-((1-(4-甲氧基苯基)-1h-苯并[d]咪唑-5-基氨基甲基)苯酚。
本申请发明人先后采用催化氢化,铁粉还原等方法进行研究将中间体Ⅰ还原制备中间体Ⅱ过程,研究中发现催化氢化采用价格昂贵且易燃的还原剂,同时需要加压反应釜,过程不易监控,有可能催化不完全。采用铁粉还原方法,反应虽然较易进行,但反应过程中产生的铁泥很难处理。最终才确定采用保险粉还原法,和现有技术相比,本发明采用保险粉还原法,反应条件温和,反应现象明显(反应液颜色变化明显,深红色最终变为淡黄色),过程易于监控,后续处理简单,溶剂无水乙醇在后处理过程中均可回收套用,可以有效降低成本,且在S4步骤中采用无水乙醇加活性炭脱色后重结晶,所得产品纯度高,外观好,后续反应副产物少。
进一步,所述S1步骤中,第二残留物和石油醚/乙酸乙酯混合溶液的质量比为1:10-1:20。
进一步,所述石油醚/乙酸乙酯混合溶液中,石油醚和乙酸乙酯的体积比为3:1-4:1。
本发明和现有的采用硅胶柱层析分离相比,本发明采用乙酸乙酯溶解反应物,再用饱和碳酸氢钠洗涤除去盐分,有机层蒸干后,石油醚与乙酸乙酯重结晶,收率较高,同时大大降低操作难度的,减少操作时间,适合于规模化制备和生产。
进一步,所述中间体Ⅰ和无水乙醇的质量比为1:20-1:40。进一步,所述S3步骤中,第五残留物与无水乙醇的体积比是1:8-1:10,所述脱色活性炭的用量为第五残留物质量的5%-10%。
进一步,所述S4步骤中,第七残留物与乙酸乙酯/环己烷混合液的体积比是1:8-1:15,所述乙酸乙酯/环己烷混合液中乙酸乙酯和环己烷的体积比为1:2-1:5。
本发明的有益效果:
1、本发明公开了一种具有抗拉沙病毒活性的化合物,为首次报道,对沙粒病毒感染,尤其是拉沙病毒感染及其相关疾病具有治疗效果,可以用于制备抗拉沙病毒的药物。
2、本发明的一种具有抗拉沙病毒活性的化合物的制备方法,工艺简单,操作简单,质量可控,收率高,工艺平行性及重现性好,适合大规模化生产。
附图说明
图1是本发明式Ⅰ化合物的高效液相色谱图。
图2是本发明式Ⅰ化合物的高分辨质谱图。
图3分别是本发明式Ⅰ化合物的核磁共振氢谱图。
图4分别是本发明式Ⅰ化合物的在假病毒模型中抑制病毒感染的检测结果
具体实施方式
以下将结合具体实施例对本发明进行详细说明:
本发明提供了一种具有抗拉沙病毒活性的化合物,即2,6-二甲氧基-4-((1-(4-甲氧基苯基)-1h-苯并[d]咪唑-5-基氨基甲基)苯酚,其结构式如式(I)所示:
其合成路线如下所示:
具有结构式(I)所示的化合物的药学上可接受的盐,可以通过加入药学上可以接受的酸来制备,可以是无机酸或有机酸。无机酸的盐包括盐酸盐、氢溴酸盐、硫酸盐、硝酸盐、磷酸盐等,有机酸的盐包括乙酸盐、丙酸盐、羟基乙酸盐、丙酮酸盐、草酸盐、苹果酸盐、丙二酸盐、琥珀酸盐、马来酸盐、富马酸盐、酒石酸盐、柠檬酸盐、苯甲酸盐、肉桂酸盐、扁桃酸盐、甲磺酸盐、乙磺酸盐、对甲苯磺酸盐、水杨酸盐等。
以下结合实施例对发明作进一步详细说明:
实施例1
S1:中间体Ⅰ(2,4-二硝基苯基-4-甲氧基苯胺)的合成。
向反应瓶中加入2,4-二硝基氟苯37.2g、对甲氧基苯胺24.6g,再加入四氢呋喃300mL,搅拌溶解后,加入碳酸铯78.2g,加热至48℃,恒温反应10-12h薄层检测反应完毕。将反应液过滤,滤液减压蒸干,得到第一残留物,向第一残留物中加入乙酸乙酯600mL搅拌至完全溶解后,加入饱和碳酸氢钠洗涤二次,每次加入饱和碳酸氢钠300mL,有机层用无水硫酸镁干燥过夜;过滤,滤液减压蒸干得到第二残留物,向第二残留物中加入15倍第二残留物质量的石油醚/乙酸乙酯混合溶液进行重结晶,得中间体Ⅰ46.3g;石油醚/乙酸乙酯混合溶液中石油醚和乙酸乙酯的体积比为3:1。
S2:中间体Ⅱ(N1-(4-甲氧基苯基)-1,2,4-三苯胺)的合成。
反应瓶中加入中间体Ⅰ46.3g,加入无水乙醇2000mL搅拌,加热至回流使之溶解;将保险粉254g用1000mL水搅拌溶解后,滴加至反应瓶中,0.5-1h滴完;滴加过程中,反应液由深红色逐渐变为浅红色,至反应完毕,最终变为淡黄色;反应液继续搅拌20min,静置,冷却至室温,过滤,滤液减压浓缩得到第三残留物,第三残留物加无水乙醇1000mL,加热搅拌20min,过滤除去不溶性盐,滤液减压浓缩,即得中间体Ⅱ31.0g,为棕色膏状固体。
S3:中间体Ⅲ(1-(4-甲氧基苯基)-5-氨基-苯并咪唑)的合成
反应瓶中加入中间体Ⅱ31.0g,加入4N盐酸600mL,搅拌使其溶解,加入无水甲酸37g,反应液加热至回流,保温反应3-5h,至反应完毕,减压蒸干溶剂,得到第四残留物,第四残留物加水500mL溶解,用5N氢氧化钠溶液调节pH值至13,用二氯甲烷800mL分两次萃取,每次使用400mL,收集有机层,加入无水硫酸镁干燥过夜,过滤,减压蒸干溶剂,得到第五残留物用无水乙醇300mL溶解,加入活性炭5.0g脱色,加热至回流20min,趁热过滤,滤液浓缩至150mL,放置于0-4℃冷却析晶,过滤,干燥,得中间体Ⅲ26g。
步骤4:化合物(2,6-二甲氧基-4-((1-(4-甲氧基苯基)-1h-苯并[d]咪唑-5-基氨基甲基)苯酚)的合成
三口反应瓶中加入中间体Ⅲ0.478g,丁香醛(3,5-二甲氧基-4-羟基苯甲醛)0.362g加入1,2-二氯乙烷40mL,搅拌使之溶解,加入醋酸0.24g,加热至60℃,搅拌3-5h,TLC监测反应进程,至反应物大部分生成亚胺中间体,等量分三批共加入三乙酰氧基硼氢化钠0.84g,于60℃反应5-7h,至反应完毕,反应液用饱和碳酸氢钠洗涤2次,每次使用40mL,分出有机层,加入无水硫酸镁干燥,过滤,减压蒸干溶剂,得到第六残留物以乙酸乙酯作为洗脱剂,通过柱色谱分离,采用乙酸乙酯和甲醇3:1为展开剂,根据薄层色谱监测情况收集斑点为Rf=0.5-0.6的组分,,减压蒸馏除去溶剂,得到第七残留物,第七残留物加入10倍第七残留物体积的乙酸乙酯/环己烷混合液,其中乙酸乙酯和环己烷的体积比为1:2,放置于4℃处冷却析晶,将析出固体过滤,干燥,即得具有抗拉沙病毒活性的化合物。
对制备得到的产物,通过LC-MS和1H-NMR进行鉴定并通过HPLC证实纯度,检测结果如图1-图3所示。
本发明所设计的高分辨质谱检测仪器为:BRUKER公司串联傅里叶质谱。
本发明所涉及的化合物及检测方法为:高效液相色谱法。色谱条件及系统适用性:色谱柱:Waters Sunfire C18(4.6×200mm,5μm),检测波长为240nm,流动相为甲醇-0.5%磷酸二氢铵(83:17,v/v),柱温40℃;流速1mL/min,进样量10μL。取对照品溶液作为系统适用性试验溶液,注入液相色谱仪测试,理论板数按主峰计算不得低于5000,主峰与相邻杂质峰的分离度应符合规定。
检测得出,本发明所得化合物的HPLC纯度:95.3%。
熔点:115.7-118.1℃。
质谱数据:ESI Postive:406.1762(M+1)为分子离子峰。
本发明的核磁检测仪器及检测条件为,VARIAN公司,INOVA 600MHz核磁共振仪,检测条件为:脉冲序列:presat,温度:27.0℃,延迟:2.000秒,时间:2.185秒,宽度:7163.3Hz,数据处理:线宽0.1Hz,总时间:1分7秒。
核磁数据:
1H-NMR(600MHz),CDCl3:δ7.917(d,1H,J,=3.6),7.377(t,1H,J1=J2=1.8),7.366(t,1H,J1=1.8,J2=3.6),7.244(d,1H,J=6.0),7.068(d,1H,J=2.4),7.037(t,1H,J1=1.8,J2=1.2),7.025(t,1H,J1=2.4,J2=3.0),6.708(dd,1H,J=6.0),6.651(s,2H),6.482(s,1H),5.407(d,1H),4.528(s,1H),4.284(s,2H),3.864(s,9H)。
实施例2测定本发明化合物的抗病毒活性
本发明构建了基于拉沙热病毒的包膜糖蛋白的假病毒模型,采用得自拉沙热病毒的包膜糖蛋白,而非采用病毒自身,因此可以在正常的BSL-2条件下安全进行。病毒进入步骤是开发抗病毒药物的重要靶标,因为病毒进入步骤是每个病毒生命周期的必要过程。
使用病毒假模标本来评价拉沙热包膜功能,通过使用荧光素酶报道基因进行拉沙病毒包膜和复制缺陷HIV原病毒的共转染产生所述病毒假模标本。原病毒经过工程化以便HIV包膜不被表达,因此当出芽病毒粒子非特异性捕获细胞表面蛋白时获得异源病毒包膜蛋白。用这样的方式制备的假模标本将通过异源包膜感染细胞并常被用于试验异源包膜的功能。通过从一体化HIV报道基因构建物产生的荧光素酶信号来测量感染。用于感染细胞培养系的传染性病毒的量与在被感染细胞中产生的由荧光素酶介导的发光在若干数量级上成正比。
本发明采用沙粒病毒GP基因(LASV-Josiah GP),经密码子优化后进行合成,所合成的基因克隆至载体pIRES2-EGFP;携带荧光素酶报告基因的HIV-1质粒(pNL4-3.Luc.R-E-)以及人胚胎肾上皮细胞HEK293T。
HEK 293FT细胞的培养和假病毒的制备
用含10%胎牛血清的DMEM培养基培养293FT细胞,于转染前1d铺于24孔板,细胞密度约70~80%进行转染。转染试剂采用稳定、方便的脂质体,操作方法参照Invitrogen公司的转染试剂LipofectAMINE2000使用说明。转染后放入含5%CO2的37℃孵箱中培养;培养2d收假病毒培养上清。
假病毒的感染
HEK 293FT细胞收集细胞培养上清,3000rpm离心5min,上清用22μm滤器过滤,-70℃储存备用。用24孔细胞培养板培养293FT细胞,待细胞密度达到60%汇合时吸去上清,加入150μL新鲜的细胞培养液和150μL上述制备的假病毒液,放置在含5%CO2的孵箱中于37℃继续培养,12h后补加新鲜的细胞培养液500μL继续培养。
荧光素酶活性分析
假型慢病毒上清感染HEK 293FT细胞2d后收集细胞,用PBS漂洗,3000rpm离心5min后弃上清,在细胞沉淀中加入1×裂解缓冲液(CCLR,Promega公司)90μL裂解细胞,吹打充分混匀,离心后取上清10μL,分别加入白色96孔板,每孔加入50μL荧光素酶检测底物,立即用检测仪检测荧光素酶活性。
化合物活性测试将HEK293T细胞按每孔2×104个接种在48孔板中,并继续培养24h。感染前10min将式Ⅰ化合物配制成不同浓度加至细胞,以等量DMSO作为溶剂对照,再加入病毒。48h后,弃去上清,裂解细胞,测定细胞裂解液中的相对荧光素酶活性,以DMSO溶剂孔RLUs值为100%,计算加药孔的感染率。
将得到的实验数据,用GraphPad prism及excel软件分析实验数据并做图,以化合物浓度-感染率作散点图转化为对数后进行线性拟合得到LASV-GPC的抑制曲线,得到其相关性、相关系数及半数抑制有效浓度(IC50=53.7nmol·L-1,R2=0.9551),结果如图4所示,可以看出,式Ⅰ化合物显著抑制了LASV-GPC的活性,且呈剂量依赖关系,由细胞实验的结果可知,本发明的化合物具有很好的抗沙粒病毒感染的活性。
本发明还提供了一种药物组合物,药物组合物包括有效剂量的一种或多种式(I)表示的化合物或其药学可接受的盐和药学可接受的载体或赋形剂。其中,药学可接受的载体”包括但不限于已经被国家药品食品管理局认可的而可用于人类或动物的任何佐剂、载体、赋形剂、助流剂、甜味剂、稀释剂、防腐剂、染料/着色剂、香味增强剂、表面活性剂、润湿剂、分散剂、助悬剂、稳定剂、等渗压剂、溶剂或乳化剂等对药物组合物无副作用的各种形式的载体。
本发明的药物组合物被制成合适的剂型,包括但不限于片剂、胶囊剂、锭剂、丸剂、颗粒剂、粉末剂、溶液剂、乳剂、混悬剂、分散体、糖浆剂、凝胶剂、或气溶胶剂。
实施例3本发明化合物的药物组合
使用以下成分制备了片剂。
将组分掺混并压缩成片,每片重250mg。
以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的宗旨和范围,其均应涵盖在本发明的权利要求范围当中。本发明未详细描述的技术、形状、构造部分均为公知技术。
Claims (10)
3.权利要求1所述的化合物或其药学上可接受的盐或权力要求2所述的药物组合物在治疗或预防哺乳动物的与沙粒病毒相关的疾病或疾病状态的药物中的应用,其中抗沙粒病毒被认为是有益的。
4.根据权利要求3所述的应用,所述哺乳动物为人。
5.根据权利要求4所述的应用,其特征在于,所述沙粒病毒为拉沙病毒、呼宁病毒、马丘波病毒、瓜纳瑞托病毒、萨比亚病毒、白水阿罗约病毒、查帕雷病毒、LCMV、LCMV样病毒中任意一种。
6.一种如权利要求1所述的具有抗拉沙病毒活性的化合物的制备方法,其特征在于,所述制备方法是以2,4-二硝基氟苯和对甲氧基苯胺为原料,在催化剂作用下反应得到中间体Ⅰ,将中间体Ⅰ经保险粉还原后得到中间体Ⅱ,再与甲酸反应后得到中间体Ⅲ,最后以中间体Ⅲ和丁香醛为原料,在三乙酰氧基硼氢化钠作用下反应得到产品2,6-二甲氧基-4-((1-(4-甲氧基苯基)-1h-苯并[d]咪唑-5-基氨基甲基)苯酚。
7.根据权利要求6所述的一种具有抗拉沙病毒活性的化合物的制备方法,其特征在于,所述中间体Ⅰ经保险粉还原制得中间体Ⅱ具体操作为:将中间体Ⅰ加入无水乙醇中,搅拌,加热至回流使其溶解,滴加保险粉水溶液,滴加完成后,持续保温至反应完成,反应液继续搅拌20min,静置,冷却至室温过滤,滤液减压浓缩,加入无水乙醇,加热搅拌20min,过滤,滤液减压浓缩,得到中间体Ⅱ。
8.根据权利要求7所述的一种具有抗拉沙病毒活性的化合物的制备方法,其特征在于,所述中间体Ⅰ和保险粉的质量比为1:6-1:9.1。
9.根据权利要求6-8任一权利要求所述的一种具有抗拉沙病毒活性的化合物的制备方法,其特征在于,所述制备方法具体包括以下步骤:
S1:将2,4-二硝基氟苯与对甲氧基苯胺加入反应釜中,再加入四氢呋喃,搅拌溶解后,加入催化剂碳酸铯,加热至48℃,保温反应10-12h,反应完成后,反应液过滤,滤液经减压蒸干后,得到第一残留物,将第一残留物加入乙酸乙酯搅拌至完全溶解后,加入饱和碳酸氢钠洗涤两次,有机层用无水硫酸镁干燥过夜,过滤、滤液减压蒸干,得到第二残留物,第二残留物加入石油醚/乙酸乙酯混合溶液进行重结晶,得到中间体Ⅰ;
S2:将中间体Ⅰ加入无水乙醇中,搅拌,加热至回流使其溶解,滴加保险粉水溶液,滴加完成后,持续保温至反应完成后,反应液继续搅拌20min,静置,冷却至室温过滤,滤液减压浓缩,得到第三残留物,第三残留物中加入无水乙醇,加热搅拌20min,过滤,滤液减压浓缩,得到中间体Ⅱ;
S3:将中间体Ⅱ加入4N盐酸搅拌溶解后,加入无水甲酸加热至回流,保温反应3-5h后,反应完成,减压蒸干溶剂,得到第四残留物加水溶解,调节pH至13,用二氯甲烷萃取后,收集有机层,有机层用无水硫酸镁干燥过夜,过滤后减压蒸干溶剂,得到第五残留物用无水乙醇溶解后,加入活性炭脱色,加热回流20min,趁热过滤,滤液浓缩至1/2体积,于0-4℃温度下冷却结晶,过滤、干燥得到中间体Ⅲ;
S4:将中间体Ⅲ、丁香醛加入1,2-二氯乙烷中,搅拌溶解后,加入醋酸,加热至60℃,搅拌3-5h,分批加入三乙酰氧基硼氢化钠,于60℃反应5-7h,反应液用饱和碳酸氢钠洗涤,有机层干燥,过滤,减压蒸干,得到第六残留物用通过硅胶快速柱色谱分离,收集目标组分,减压蒸馏除去溶剂,得到第七残留物加入乙酸乙酯/环己烷混合液,放置于4℃处冷却析晶,过滤,干燥,得到目的物,即2,6-二甲氧基-4-((1-(4-甲氧基苯基)-1h-苯并[d]咪唑-5-基氨基甲基)苯酚。
10.根据权利要求9所述的一种具有抗拉沙病毒活性的化合物的制备方法,其特征在于,所述S1步骤中采用石油醚/乙酸乙酯混合溶液作为重结晶液,所述石油醚和乙酸乙酯的体积比为3:1-4:1。
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Title |
---|
KARL J. OKOLOTOWICZ ET AL.,: "1,5-Disubstituted benzimidazoles that direct cardiomyocyte differentiation from mouse embryonic stem cells", 《BIOORGANIC & MEDICINAL CHEMISTRY》 * |
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