CN110066248B - 一种制备中乌宁的方法及其相关中间体 - Google Patents
一种制备中乌宁的方法及其相关中间体 Download PDFInfo
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Abstract
本发明涉及制备中乌宁的方法及其相关中间体。具体地,本发明的制备中乌宁的方法以乌头属植物准噶尔乌头提取的乌头碱为原料经碱化、乙酰化、N‑去乙基、N‑甲基化和水解得到中乌宁。本发明的制备中乌宁的方法工艺简单,收率高,环保无污染,适合工业化生产。
Description
技术领域
本发明涉及合成领域,具体涉及制备中乌宁(mesaconine)的方法及其相关中间体。
背景技术
中药附子为毛茛科植物乌头(Aconitum carmichaeli Debx.)的子根的加工品,具有回阳救逆,逐风寒湿邪,温经止痛功效(中国药典,2015年版一部),作为著名的中药广泛用于临床。
研究发现中乌宁是附子的主要强心活性成分,如在以下文献中有相关记载:Xiu-Xiu Liu,et al,Chem.Pharm.Bull,2012,60(1),144-149;Xi-Xian Jian,et al,Nat.Prod.Commun,2012,7(6),713-720;中国发明专利,2012年,CN 102146057B;Zhong-Tang Zhang,et al,Nat.Prod.Commun,2015,10(12),2075-2084。因此,中乌宁具有开发成抗心衰药物的前景。
中乌宁的化学名称为N-甲基-1α,6α,16β,18-四甲氧基-4-甲基-乌头-3α,8β,13β,14α,15α–五醇,分子式为C24H39NO9,Cas No.:6792-09-2,具有如下结构式。
中国发明专利CN 102146057B公开了中乌宁的制备方法,以分自乌头属植物准噶尔乌头(Aconitum soongaricum Stapf.)的乌头碱为原料,经全乙酰化,N-去乙基化,N-甲基化和水解得到中乌宁。然而,该制备方法涉及剧毒性的乌头碱(aconitine)和中乌头碱(mesaconitine),不利于安全生产。
为了克服现有技术的不足,有必要开发一种操作简单、收率高、环保无污染、适合工业化的中乌宁的制备工艺。
发明内容
本发明的目的在于提供一种中乌宁的制备方法,该方法操作简单、收率高、环保无污染,适合工业化生产。
本发明的中乌宁的制备方法包括以下步骤:
1)将乌头碱(I)溶于乙醇(如,95%乙醇),加入氢氧化钠水解,减压浓缩,残留物用水稀释,二氯甲烷萃除杂质生物碱,碱水液盐酸酸化,氨水或氢氧化钠乙醇溶液调至pH11~12,减压浓缩至干,残留物用二氯甲烷-乙醇(6:1至12:1,优选9:1,V/V)溶解,过滤,滤液减压浓缩,得乌头宁碱(II);
2)乌头宁碱(II)溶于吡啶中,与醋酐反应,减压浓缩得残留物,加水稀释,氨水碱化,二氯甲烷萃取,合并二氯甲烷层,依次经干燥,浓缩,硅胶柱层析分离,制得3,14,15-三乙酰乌头宁碱(III);
或者
乌头宁碱(II)在对甲苯磺酸催化下,与醋酐反应,减压浓缩得残留物,加水稀释,氨水碱化,二氯甲烷萃取,合并二氯甲烷层,依次经干燥,浓缩,硅胶柱层析分离,制得3,8,13,14,15-五乙酰乌头宁碱(IV);
3)3,14,15-三乙酰乌头宁碱(III)溶于冰醋酸中,加入N-溴代丁二酰亚胺,室温搅拌,减压浓缩得残留物,加入氨水,用二氯甲烷萃取,合并二氯甲烷萃取液,经干燥,减压浓缩,制得N-去乙基-3,14,15-三乙酰乌头宁碱(V);
或者
3,8,13,14,15-五乙酰乌头宁碱(IV)溶于冰醋酸中,加入N-溴代丁二酰亚胺,室温搅拌,减压浓缩得残留物,加入氨水,用二氯甲烷萃取,合并二氯甲烷萃取液,经干燥,减压浓缩,制得N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI);
4)N-去乙基-3,14,15-三乙酰乌头宁碱(V)溶于四氢呋喃中,室温下加入甲醛水溶液和冰醋酸,室温搅拌,加入NaBH(OAc)3,继续搅拌,加入氨水(调至pH9-10),加水稀释后,用二氯甲烷萃取,合并二氯甲烷萃取液,依次经水洗,干燥,减压浓缩,制得3,14,15-三乙酰中乌宁(VII);
或者
N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI)溶于四氢呋喃中,室温下加入甲醛水溶液和冰醋酸,室温搅拌,加入NaBH(OAc)3,继续搅拌,加入氨水(调至pH9-10),加水稀释后,用二氯甲烷萃取,合并二氯甲烷萃取液,依次经水洗,干燥,减压浓缩,制得3,8,13,14,15-五乙酰中乌宁(VIII);
5)3,14,15-三乙酰中乌宁(VII)或3,8,13,14,15-五乙酰中乌宁(VIII)溶于乙醇(如,95%乙醇)溶液中,加入氢氧化钠反应,盐酸调至pH4-5,再用氨水或氢氧化钠乙醇溶液调至pH9~12,滤去不溶物,减压浓缩,残留物用二氯甲烷-乙醇(6:1至12:1,优选9:1,V/V)溶解,抽滤,滤液减压浓缩,制得中乌宁(IX)
在本发明的实施方案中,所述乌头碱(I)可以通过以下方法制备:
将乌头属植物的根粉碎后,加硫酸-乙醇水溶液回流提取;提取液减压浓缩,得固体提取物;固体提取物用水稀释,碱化后用醋酸乙酯萃取,回收溶剂,得醋酸乙酯提取物;醋酸乙酯提取物经酸溶、过滤、碱化沉淀,得所述乌头碱(I)。
在本发明的实施方案中,基于所述硫酸-乙醇水溶液的总质量,硫酸的含量可以为1-10‰,乙醇的含量可以为80-90%,优选地,硫酸的含量可以为5‰,乙醇的含量可以为85%。
在本发明的实施方案中,所述乌头属植物可以为准噶尔乌头或多根乌头。
更进一步地,本发明的中乌宁通过以下步骤制备获得:
1)将乌头属植物准噶尔乌头的根粉碎后,加5‰硫酸85%乙醇水溶液回流提取;提取液减压浓缩,得固体提取物;固体提取物用水稀释,碱化后用醋酸乙酯萃取,回收溶剂,得醋酸乙酯提取物;醋酸乙酯提取物经酸溶、过滤、碱化沉淀,得总生物碱,其中总生物碱中主要成分为乌头碱(I);
2)将所述总生物碱溶于95%乙醇,加入所述总生物碱中乌头碱(I)总量的3~5倍摩尔量的氢氧化钠,室温反应1~2小时,减压浓缩除去95%乙醇,残留物用5~10倍量水稀释,二氯甲烷萃取除去杂质生物碱,碱水液用盐酸调至pH 5,氨水或氢氧化钠乙醇溶液调至pH11~12,减压浓缩至干,残留物用二氯甲烷-无水乙醇(9:1,V/V)溶解,过滤,滤液减压浓缩,得乌头宁碱(II);
3)乌头宁碱(II)溶于吡啶中,乌头宁碱(II)与6~8摩尔醋酐回流反应2-3小时,减压浓缩得残留物,加水稀释,氨水碱化,二氯甲烷萃取,合并二氯甲烷层,依次经无水硫酸钠干燥,浓缩,硅胶柱层析(二氯甲烷-乙醇=200:1)分离,得3,14,15-三乙酰乌头宁碱(III);
或者
乌头宁碱(II)在对甲苯磺酸催化下,乌头宁碱(II)与6~8摩尔醋酐回流反应3-4小时,减压浓缩得残留物,加水稀释,氨水碱化,二氯甲烷萃取,合并二氯甲烷层,依次经无水硫酸钠干燥,浓缩,硅胶柱层析(石油醚-丙酮(10:1~2:1)洗脱)分离,得3,8,13,14,15-五乙酰乌头宁碱(IV);
4)3,14,15-三乙酰乌头宁碱(III)溶于10倍量冰醋酸中,与3~5摩尔的N-溴代丁二酰胺室温搅拌反应2-3小时,减压浓缩得残留物,浓氨水调至pH9~10,用二氯甲烷萃取2次,合并二氯甲烷萃取液,无水硫酸钠干燥,减压浓缩,得N-去乙基-3,14,15-三乙酰乌头宁碱(V);
或者
3,8,13,14,15-五乙酰乌头宁碱(IV)溶于10倍量冰醋酸中,与3~5摩尔的N-溴代丁二酰胺室温搅拌反应2-3小时,减压浓缩得残留物,浓氨水调至pH9~10,用二氯甲烷萃取2次,合并二氯甲烷萃取液,无水硫酸钠干燥,减压浓缩,得N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI);
5)N-去乙基-3,14,15-三乙酰乌头宁碱(V)溶于四氢呋喃中,与12摩尔40%甲醛水溶液和1~2摩尔冰醋酸室温搅拌反应30~60分钟后,加入2~3摩尔三乙酰氧基硼氢化钠,室温搅拌反应30~60分钟,浓氨水调至pH9-10,加水稀释后,用二氯甲烷萃取,合并二氯甲烷萃取液,依次经,无水硫酸钠干燥,减压浓缩,得3,14,15-三乙酰中乌宁(VII);
或者
N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI)溶于四氢呋喃中,与12摩尔40%甲醛水溶液和1~2摩尔冰醋酸室温搅拌反应30~60分钟后,加入2~3摩尔三乙酰氧基硼氢化钠,室温搅拌反应30~60分钟,浓氨水调至pH9-10,加水稀释后,用二氯甲烷萃取,合并二氯甲烷萃取液,依次经无水硫酸钠干燥,减压浓缩,得3,8,13,14,15-五乙酰中乌宁(VIII);
6)3,14,15-三乙酰中乌宁(VII)或3,8,13,14,15-五乙酰中乌宁(VIII)用5%~10%氢氧化钠乙醇液水解,浓盐酸调至pH4-5,再用稀氢氧化钠乙醇溶液调至pH9~12,滤去不溶物,减压浓缩,残留物用二氯甲烷/无水乙醇(9:1,V/V)溶解,抽滤,滤液减压浓缩,得中乌宁(IX)
本发明还提供了制备中乌宁的方法中使用的中间体,具体如下:
N-去乙基-3,14,15-三乙酰乌头宁碱,其结构式如式(V)所示
N-去乙基-3,8,13,14,15-五乙酰乌头宁碱,其结构式如式(VI)所示
3,14,15-三乙酰中乌宁,其结构式如式(VII)所示
3,8,13,14,15-五乙酰中乌宁,其结构式如式(VIII)所示
总生物碱中的乌头碱具有一定毒性,本发明的制备中乌宁的方法通过首先将总生物碱中的乌头碱(I)中的苯甲酰基水解得到无毒性的式II中间体(乌头宁碱),再依次经乙酰化、N-去乙基、N-甲基化分别得到无毒性的式III中间体(3,14,15-三乙酰乌头宁碱)、式IV中间体(3,8,13,14,15-五乙酰乌头宁碱)、式V中间体(N-去乙基-3,14,15-三乙酰乌头宁碱)、式VI中间体(N-去乙基-3,8,13,14,15-五乙酰乌头宁碱)、式VII中间体(3,14,15-三乙酰中乌宁)和式VIII中间体(3,8,13,14,15-五乙酰中乌宁),从而避免了毒性中间体的使用,保证生产安全。本发明的中乌宁制备方法中使用的上述中间体不仅无毒性,而且使得工艺操作简单,纯化效果好。
具体实施方案
下列实施例进一步解释说明本发明,但是,它们并不构成对本发明范围的限制或限定。
实施例1乌头属植物准噶尔乌头总生物碱的提取
(1)取10kg乌头属植物准噶尔乌头干燥根,粉碎,过20目筛;
(2)乌头属植物准噶尔乌头粉末依次用其80L、36L和24L倍量的5‰硫酸85%乙醇水溶液回流提取3次,每次提取2小时,过滤,合并滤液;
(3)滤液减压浓缩,回收乙醇至流浸膏相对密度1.05~1.10,收集流浸膏0.46kg;
(4)流浸膏加1.4L水稀释,用氨水碱化,调至pH 10,放入萃取器中用乙酸乙酯萃取(2L×3次),每次搅拌5~10分钟,收集萃取液;
(5)减压浓缩回收乙酸乙酯,得总生物碱129g。取样HPLC法测定总生物碱中乌头碱的含量。由其含量和湿重计算出总生物碱中乌头碱(I)的总量约为42.5g,收率约0.42%。
实施例2中乌宁的制备
(1)乌头宁碱(II)的制备
取100g总生物碱(含乌头碱33g)用500ml 95%乙醇溶解后,加入7.2g(78mmol)氢氧化钠,室温下搅拌反应2小时,减压回收溶剂得120g固体物。固体物中加入1000ml水稀释后,用二氯甲烷萃取(500ml×2次)。水层用浓盐酸调至pH 5,再用稀氢氧化钠乙醇溶液调至pH11-12,减压浓缩至干,得固体物90g,加入二氯甲烷-无水乙醇(9:1,V/V)900ml,加热溶解,过滤,滤液减压浓缩得固体物,得所需化合物22.5g。
收率:86%,白色无定形粉末,C25H41NO9。
1H NMR(400MHz,CD3OD)δ:0.99(3H,t,NCH2CH3),3.15,3.20,3.25,3.50(各3H,s,4xOCH3),3.59(1H,dd,J=12.0,4.0Hz,H-3β),3.71(1H,d,J=3.77(1H,d,J=8.0Hz),4.09,(1H,d,J=8.0Hz,H-15β),4.32(1H,d,J=4.0Hz,H-14β);
13C NMR(100MHz,CD3OD)δ:13.8(q),35.5(t),38.8(t),43.1(d),44.5(s),47.5(d),48.3(t),49.5(d),50.1(t),50.3(d),51.1(s),56.2(q),58.2(q),59.1(q),61.6(q),62.1(d),70.7(d),75.3(t),77.7(s),79.5(s),79.7(d),82.3(d),84.2(d),85.0(d),93.1(d);
ESI-MS m/z(%):500(100)[M+H]+。
(2)3,14,15-三乙酰乌头宁碱(III)的制备
取10.0g(20mmol)乌头宁碱(II)与7.1g(70mmol)摩尔醋酐和100ml吡啶混合,在回流条件下反应2.5小时,减压浓缩得残留物,用170ml水稀释残留物,氨水碱化至pH9~10,二氯甲烷萃取(80ml×3),合并萃取液,无水硫酸钠干燥,减压浓缩得固体物,硅胶柱层析,二氯甲烷-无水乙醇(200:1)洗脱,得所需化合物10.7g。
收率:85.6%,白色无定形粉末,C31H47NO12。
1H NMR(400MHz,CDCl3)δ:1.13(3H,t,J=7.2Hz,NCH2CH3),2.06,2.07,2.18(各3H,s,3XOAc),3.20,3.23,3.27,3.56(各3H,s,4XOCH3),4.65(1H,d,J=5.2,H-14β),4.88(1H,t,J=8.8Hz,H-3β),5.25(d,J=6.0Hz,H-15β);
13C NMR(100MHz,CDCl3)δ:13.4(q),20.8(q),21.1(q),21.1(q),31.9(t),36.2(t),40.6(d),42.0(s),44.5(d),45.5(d),47.4(t),48.9(t),49.5(d),49.5(s),56.1(q),57.8(q),58.7(q),60.6(d),61.3(q),71.4(d),71.7(t),74.3(s),76.5(s),78.6(d),81.7(d),82.8(d),87.2(d),88.3(d),170.1(s),170.9(s),173.3(s);
ESI-MS m/z(%):626(100)[M+H]+。
(3)3,8,13,14,15-五乙酰乌头宁碱(IV)的制备
取150mg(0.31mmol)乌头宁碱(II)溶于1mL醋酐中,室温搅拌下加入176mg(0.93mmol)对甲基苯磺酸。将反应液升温至120℃,反应3~4小时,减压浓缩蒸除大部分溶剂,所得残留液以20mL水稀释,5mL氨水调至pH 9~10,二氯甲烷萃取(10mL×2),合并萃取液,无水硫酸钠干燥,抽滤,减压浓缩得残留液,硅胶柱层析分离,石油醚-丙酮(10:1~2:1)洗脱,得所需化合物161mg。
收率75%,白色无定形粉末,C35H51NO14。
1H NMR(400MHz,CDCl3)δ:1.14(3H,t,J=7.2Hz,NCH2CH3),1.94(3H,s,OAc),2.04,2.08(各3H,s,2x OAc),2.10,2.13(各3H,s,2x OAc),3.19,3.21,3.25,3.33(各3H,s,4xOCH3),4.05(1H,d,J=6.0Hz,H-6β),4.84(1H,d,J=5.2Hz,H-14β),4.88(1H,dd,J=12.4,6.8Hz,H-3β),5.84(1H,d,J=6.0Hz,H-15β);
13C NMR(100MHz,CDCl3)δ:13.5(q),21.1(q),21.1(q),21.2(q),21.3(q),22.0(q),31.9(t),36.0(t),41.1(d),42.1(s),43.7(d),45.0(d),45.5(d),47.0(t),48.8(t),49.8(s),56.1(q),58.7(q),58.8(q),60.3(q),61.0(d),71.2(t),71.3(d),76.3(d),78.2(d),80.7(s),81.4(d),83.7(d),88.2(d),88.8(s),168.4(s),169.6(s),170.2(s),170.3(s),170.7(s);
ESI-MS m/z(%):710(100)[M+H]+。
(4)N-去乙基-3,14,15-三乙酰乌头宁碱(V)的制备
取10.0g(17mmol)3,14,15-三乙酰乌头宁碱(III),溶于100ml冰醋酸中,加入9.9g(66mmol)N-溴代丁二酰亚胺,室温搅拌反应2小时,减压浓缩得固体物,用少量二氯甲烷溶解,加入150ml水,浓氨水碱化至pH10,用二氯甲烷萃取(80ml×2)。合并萃取液,无水硫酸钠干燥,减压浓缩得10.5g固体物,硅胶柱层析,石油醚-丙酮(2:1)洗脱,得所需化合物6.4g。
收率:67.0%,白色无定形粉末,C29H43NO12。
1H NMR(400MHz,CDCl3)δ:2.05,2.08,2.27(各3H,s,3XOAc),3.24,3.25,3.29,3.57(各3H,s,4XOCH3),4.68(1H,d,J=4.8Hz,H-14β),5.09(1H,t,J=6.4Hz,H-3β),5.25(1H,d,J=6.4Hz,H-15β);
13C NMR(100MHz,CDCl3)δ:21.0(q),21.1(q),21.2(q),30.4(t),35.1(t),40.4(d),41.4(t),42.6(s),43.3(d),44.0(d),49.3(t),54.5(s),55.2(q),56.1(q),57.8(q),58.8(q),61.4(d),72.1(d),74.2(t),74.4(s),76.7(s),78.5(d),80.3(d),82.6(d),86.8(d),87.9(d),170.1(s),170.7(s),174.2(s);
ESI-MS m/z(%):598(100)[M+H]+。.
(5)N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI)的制备
取10.0g(14mmol)3,8,13,14,15-五乙酰乌头宁碱(IV),溶于100ml冰醋酸中,加入9.9g(66mmol)N-溴代丁二酰亚胺,室温搅拌反应2小时,减压浓缩得固体物,用少量二氯甲烷溶解,加入150ml水,浓氨水碱化至pH10,用二氯甲烷萃取(80ml×2)。合并萃取液,无水硫酸钠干燥,减压浓缩得固体物8.5g。硅胶柱层析,石油醚-丙酮(2:1)洗脱,得所需化合物6.5g。
收率:67.7%,白色无定形粉末,C33H47NO14。
1H NMR(400MHz,CDCl3)δ:1.94(3H,s,OAc),2.03,2.07(各3H,s,2x OAc),2.14,2.16(各3H,s,2x OAc),3.22(3H,s,OMe),3.26(6H,s,2x OMe),3.32(3H,s,OMe),4.86(1H,d,J=5.2Hz,H-14β),5.02(1H,dd,J=9.6,5.6Hz,H-3β),5.84(1H,d,J=6.0Hz,H-15β);
13C NMR(100MHz,CDCl3)δ:21.0(q),21.1(q),21.3(q),21.3(q),22.1(q),31.6(t),34.7(t),41.1(t),41.2(d),42.7(s),42.8(d),44.5(d),49.3(s),51.4(d),55.7(q),55.8(q),58.7(q),58.8(q),61.1(d),71.8(d),73.0(t),76.3(d),78.2(d),80.5(s),80.6(d),83.8(d),87.8(d),88.4(s),168.3(s),169.7(s),170.3(s),170.4(s),170.6(s);
ESI-MS m/z(%):682(100)[M+H]+。
(6)3,14,15-三乙酰中乌宁(VII)的制备
取10g(17mmol)N-去乙基-3,14,15-三乙酰乌头宁碱(V)溶于25ml四氢呋喃中,室温下加入2ml 40%甲醛水溶液和1ml冰醋酸,保持该温度搅拌30分钟后,再加入7.1g(33.5mmol)NaBH(OAc)3,继续搅拌30分钟,浓氨水调至pH9,加15ml水稀释,用20ml二氯甲烷萃取2次,合并萃取液,依次用水洗,无水硫酸钠干燥,减压浓缩至干,得5.5g固体物,硅胶柱层析,氯仿-甲醇(9:1)洗脱,得所需化合物5.41g。
收率:53.1%,白色无定形粉末,C30H45NO12。
1H NMR(400MHz,CDCl3)δ:2.05,2.07,2.21(各3H,s,3XOAc),2.32(s,NCH3),3.20,3.25,3.27,3.54(各3H,s,4XOCH3),4.65(d,J=3.2Hz,H-14β),4.88(t,J=4.4Hz,H-3β),5.24(d,J=6.0Hz,H-15β);
13C NMR(100MHz,CDCl3)δ:20.9(q),21.0(q),21.1(q),31.9(t),36.0(t),40.7(d),42.3(s),42.5(q),44.4(d),44.8(d),48.4(d),49.5(s),49.9(t),56.5(q),57.8(q),58.7(q),61.2(q),62.0(d),72.1(d),71.5(t),74.3(s),76.3(s),78.5(d),81.8(d),82.6(d),87.2(d),88.3(d),170.1(s),170.9(s),173.4(s);
ESI-MS m/z(%):612(100)[M+H]+。
(7)3,8,13,14,15-五乙酰中乌宁(VIII)的制备
取10g(14.6mmol)N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI)溶于25ml四氢呋喃中,室温下加入2ml 40%甲醛水溶液和1ml冰醋酸,搅拌30分钟后,再加入7.1g(33.5mmol)NaBH(OAc)3,继续搅拌30分钟,浓氨水调至pH 9,加15ml水稀释,用20ml二氯甲烷萃取2次。合并萃取液,依次用水洗,无水硫酸钠干燥,减压浓缩至干,得6.5g固体物。硅胶柱层析,氯仿-甲醇(9:1)洗脱,得所需化合物6.1g。
收率:59.8%,白色无定形粉末,C34H49NO14。
1H NMR(400MHz,CDCl3)δ:1.95(3H,s,OAc),2.04,2.08(各3H,s,2x OAc),2.14(6H,s,2x OAc),2.43(3H,s,NMe),3.19,3.23,3.26,3.33(各3H,s,4x OMe),4.06(1H,d,J=6.4Hz,H-6β),4.86(1H,d,J=5.2Hz,H-14β),4.90(1H,dd,J=12.4,6.0Hz,H-3),5.83(1H,d,J=6.0Hz,H-15β);
13C NMR(100MHz,CDCl3)δ:21.1(q),21.3(q),21.3(q),22.0(q),26.9(q),31.9(t),35.8(t),41.2(q),42.4(s),42.7(d),43.8(d),44.1(d),44.8(d),49.6(t),49.9(s),56.4(q),58.7(q),58.8(q),60.9(q),61.9(d),71.1(t),71.2(d),76.3(d),78.3(d),80.7(s),81.5(d),83.6(d),88.2(d),88.7(s),168.4(s),169.5(s),170.1(s),170.2(s),170.7(s);
ESI-MS m/z(%):696(100)[M+H]+。
(8)中乌宁(IX)的制备
方法1:由3,14,15-三乙酰中乌宁(VII)制备中乌宁(IX)
取10g(16.3mmol)3,14,15-三乙酰中乌宁(VII),溶于75ml 95%乙醇溶液中,加入2.29g(57.2mmol)氢氧化钠,回流反应30min。反应液冷却至室温,浓盐酸调至pH5,再用稀氢氧化钠乙醇溶液调至pH11~12。滤去不溶物,减压浓缩,得固体物。固体物用110ml二氯甲烷-无水乙醇(9:1,V/V)溶解,抽滤,滤液减压浓缩至干,得中乌宁7.22g,收率:91.1%。
方法2:由3,8,13,14,15-五乙酰中乌宁(VIII)制备中乌宁(IX)
取10g(14.4mmol)3,8,13,14,15-五乙酰中乌宁(VIII),溶于75ml 95%乙醇溶液中,加入2.29g(50.4mmol)氢氧化钠,回流反应30min。反应液冷却至室温,浓盐酸调至pH5,再用浓氨水调至pH9。滤去不溶物,减压浓缩得固体物9.5g。固体物用95ml二氯甲烷-无水乙醇(9:1,V/V)溶解,抽滤,滤液减压浓缩至干,得中乌宁6.2g,收率:90.0%。
上述中乌宁经光谱分析,具有如下理化特性:
中乌宁的光谱数据如下:
IR(KBr):3424cm-1,2932cm-1,2892cm-1,2821cm-1,1639cm-1,1453cm-1,1106cm-1;
1H NMR(400MHz,CDCl3)δ:3.07(1H,dd,J=8.4,6.0Hz,H-1β),3.75(1H,m,H-3β),4.15(d,J=6,8Hz,H-6β),3,93(1H,d.J=5.2Hz,H-14β),4.49)(1H,t,J=5.6Hz,H-15β),3.19(1H,d,J=6.0Hz,H-16),2.91(1H,s,H-17),3.65,3.74(各1H,ABq,J=9.6Hz,H2-18),2.48,2.79(各1H,ABq,J=11.2Hz,H2-19),2.34(3H,s,NCH3),3.26(3H,s,OCH3-1),3.34(3H,s,OCH3-6),3.64(3H,s,OCH3-16),3,32(3H,s,OCH3-18);
13C NMR(100MHz,CDCl3)δ:82.6(C-1),33.8(C-2),71.5(C-3),43.4(C-4),46.5(C-5),83.2(C-6),46.4(C-7),78.8(C-8),48.8(C-9),41.6(C-10),50.1(C-11),36.9(C-12),76.3(C-13),78.7(C-14),81.5(C-15),90.7(C-16),62.5(C-17),76.8(C-18),49.8(C-19),42.5(NCH3),56.3(OCH3-1),57.9(OCH3-6),61.3(OCH3-16),59.1(OCH3-18);
ESI-MS m/z(%):486(100)[M+H]+;
HR-ESI-MS:准分子量测定值为486.2699,计算值为:486.2644。
Claims (8)
1.一种制备中乌宁的方法,包括以下步骤:
1)将乌头碱(I)溶于乙醇,加入氢氧化钠水解,减压浓缩,残留物用水稀释,二氯甲烷萃除杂质生物碱,碱水液盐酸酸化,氨水或氢氧化钠乙醇溶液调至pH11~12,减压浓缩至干,残留物用二氯甲烷-乙醇溶解,过滤,滤液减压浓缩,得乌头宁碱(II);
2)乌头宁碱(II)溶于吡啶中,与醋酐反应,减压浓缩得残留物,加水稀释,氨水碱化,二氯甲烷萃取,合并二氯甲烷层,依次经干燥,浓缩,硅胶柱层析分离,制得3,14,15-三乙酰乌头宁碱(III);
或者
乌头宁碱(II)在对甲苯磺酸催化下,与醋酐反应,减压浓缩得残留物,加水稀释,氨水碱化,二氯甲烷萃取,合并二氯甲烷层,依次经干燥,浓缩,硅胶柱层析分离,制得3,8,13,14,15-五乙酰乌头宁碱(IV);
3)3,14,15-三乙酰乌头宁碱(III)溶于冰醋酸中,加入N-溴代丁二酰亚胺,室温搅拌,减压浓缩得残留物,加入氨水,用二氯甲烷萃取,合并二氯甲烷萃取液,经干燥,减压浓缩,制得N-去乙基-3,14,15-三乙酰乌头宁碱(V);
或者
3,8,13,14,15-五乙酰乌头宁碱(IV)溶于冰醋酸中,加入N-溴代丁二酰亚胺,室温搅拌,减压浓缩得残留物,加入氨水,用二氯甲烷萃取,合并二氯甲烷萃取液,经干燥,减压浓缩,制得N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI);
4)N-去乙基-3,14,15-三乙酰乌头宁碱(V)溶于四氢呋喃中,室温下加入甲醛水溶液和冰醋酸,室温搅拌,加入NaBH(OAc)3,继续搅拌,加入氨水,加水稀释后,用二氯甲烷萃取,合并二氯甲烷萃取液,依次经水洗,干燥,减压浓缩,制得3,14,15-三乙酰中乌宁(VII);
或者
N-去乙基-3,8,13,14,15-五乙酰乌头宁碱(VI)溶于四氢呋喃中,室温下加入甲醛水溶液和冰醋酸,室温搅拌,加入NaBH(OAc)3,继续搅拌,加入氨水,加水稀释后,用二氯甲烷萃取,合并二氯甲烷萃取液,依次经水洗,干燥,减压浓缩,制得3,8,13,14,15-五乙酰中乌宁(VIII);
5)3,14,15-三乙酰中乌宁(VII)或3,8,13,14,15-五乙酰中乌宁(VIII)溶于乙醇溶液中,加入氢氧化钠反应,盐酸调至pH4-5,再用氨水或氢氧化钠乙醇溶液调至pH9-12,滤去不溶物,减压浓缩,残留物用二氯甲烷-乙醇溶解,抽滤,滤液减压浓缩,制得中乌宁(IX)
2.根据权利要求1所述的方法,其中所述乌头碱(I)通过以下方法制备:
将乌头属植物的根粉碎后,加硫酸-乙醇水溶液回流提取;提取液减压浓缩,得固体提取物;固体提取物用水稀释,碱化后用醋酸乙酯萃取,回收溶剂,得醋酸乙酯提取物;醋酸乙酯提取物经酸溶、过滤、碱化沉淀,得所述乌头碱(I)。
3.根据权利要求2所述的方法,其中基于所述硫酸-乙醇水溶液的总质量,硫酸的含量为1-10‰,乙醇的含量为80-90%。
4.根据权利要求2或3所述的方法,其中基于所述硫酸-乙醇水溶液的总质量,硫酸的含量为5‰,乙醇的含量为85%。
5.根据权利要求2所述的方法,其中所述乌头属植物为准噶尔乌头或多根乌头。
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DE19511235C2 (de) * | 1995-03-27 | 1999-09-23 | Klinge Co Chem Pharm Fab | Verwendung von Aconitinen und Aconinen aus der Eisenhutknolle zur Prophylaxe einer echten Migräne |
CN101759640B (zh) | 2008-12-25 | 2012-02-29 | 上海中药制药技术有限公司 | 高纯度苯甲酰新乌头原碱的制备方法 |
CN102146057B (zh) * | 2011-02-18 | 2012-11-07 | 四川大学 | C19-二萜生物碱及其制备方法和以该化合物为活性成分的药物组合物及用途 |
CN102977020A (zh) | 2012-12-05 | 2013-03-20 | 四川大学 | 乌头碱型生物碱及其制备方法和以该化合物为强心和抗心衰活性成分的药物组合物及用途 |
CN102973679B (zh) * | 2012-12-21 | 2014-06-11 | 四川省中医药科学院 | 乌头属药材或其加工品中总生物碱提取物的制备方法 |
CN107445893A (zh) * | 2017-08-31 | 2017-12-08 | 珠海润都制药股份有限公司 | 盐酸去甲乌药碱新晶型及其制备方法 |
CN108409657B (zh) * | 2017-11-24 | 2021-08-20 | 孙益民 | 高纯高乌甲素及其制备方法 |
CN112375080A (zh) * | 2018-01-24 | 2021-02-19 | 好医生药业集团有限公司 | 中乌宁晶型及其制备方法 |
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EP3744711B1 (en) | 2022-09-21 |
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US20210040042A1 (en) | 2021-02-11 |
KR20200108004A (ko) | 2020-09-16 |
JP2021512941A (ja) | 2021-05-20 |
CN110066248A (zh) | 2019-07-30 |
EP3744711A4 (en) | 2021-07-07 |
WO2019144888A1 (zh) | 2019-08-01 |
KR102518166B1 (ko) | 2023-04-05 |
JP7072674B2 (ja) | 2022-05-20 |
EP3744712A1 (en) | 2020-12-02 |
CN112375080A (zh) | 2021-02-19 |
US10941116B2 (en) | 2021-03-09 |
EP3744711A1 (en) | 2020-12-02 |
AU2019212668A1 (en) | 2020-08-13 |
CN110066247B (zh) | 2021-01-01 |
EA202091779A1 (ru) | 2020-10-14 |
CN110066247A (zh) | 2019-07-30 |
EP3744712A4 (en) | 2021-07-07 |
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