CN115403587B - 一类中草药肝损伤相关生物标志物的化学合成方法 - Google Patents
一类中草药肝损伤相关生物标志物的化学合成方法 Download PDFInfo
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- C—CHEMISTRY; METALLURGY
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Abstract
本发明公开了一类中草药肝损伤相关生物标志物的化学合成方法。本发明以可商购的天然吡咯里西啶生物碱为起始原料合成中草药肝损伤生物标志物二羟乙基脱氢吡咯里西啶生物碱,合成步骤包括:吡咯里西啶生物碱依次和氧化剂/催化剂反应,加入稀酸反应,收集有机相洗涤、干燥后和金属醇盐反应制备得到脱氢吡咯里西啶生物碱,然后再和路易斯酸反应制备得到目标产物。本发明以可商购的天然吡咯里西啶生物碱为起始原料,反应选择性高、且副反应少易于纯化,适合放大合成及工业化生产。本发明的合成方法大幅度提高了该生物标志物的合成收率,达到生产级别要求,为吡咯里西啶生物碱类中草药导致的肝损伤疾病的精准医疗体系的建立提供关键技术支撑。
Description
技术领域
本发明涉及一类中草药肝损伤相关生物标志物的化学合成方法,属于化学合成技术领域。
背景技术
药物性肝损伤(Drug induced liver injury,DILI)是最常见和最严重的药物不良反应之一。近年来,随着新药的不断出现和中草药的广泛应用,DILI的发病率也随之增加。2019年,全球DILI的年发病率约为14-19/10万人。中国DILI 的年发生率则达到23.80/10万人。中草药及保健品在我国使用广泛,但其所致的肝损害发病隐匿,部分患者发展至肝衰竭,已经成为我国DILI比例最大的一种病因,约为26.81%。
作为一种血管型DILI的代表,含吡咯里西啶生物碱(Pyrrolizidine alkaloids,PAs)的中草药或制剂引起的肝静脉闭塞综合征或肝窦阻塞综合征(Hepatic SinusoidalObstructive Syndrome,HSOS)病例日益增多。全球范围由PAs导致 HSOS的临床报道已超过8000例,我国以土三七为主。土三七等中草药里所含有的PAs在肝细胞代谢脱氢后产生化学性质更活泼的脱氢吡咯里西啶生物碱;该类生物碱易于血浆和细胞内会与含巯基蛋白质、谷胱甘肽、核苷碱基等亲核性物质反应,经加成反应得到一系列PAs加成蛋白、加成核酸等分子,严重影响肝细胞正常生理功能,由此产生肝毒性和肝损伤。伴随病程的进一步发展,将导致肝窦和肝小静脉的内皮细胞损伤和坏死,肝脏血液“只进不出”,引起门静脉压力增加,形成腹水、黄疸等症状。这种损害一经形成,常常无法逆转,并快速发展为肝功能衰竭,危及患者生命。本团队总结国内报道的116例PAs导致的 HSOS,发现9.7%患者病情迅速进展最终死于多器官功能衰竭。
PA-HSOS的诊断主要是排除性诊断,常被误诊为Budd-Chiari综合征、失代偿期肝硬化或急性重型肝炎等。临床常用的Seattle和Baltimore诊断标准,均基于体重增加(腹水)、肝肿大以及黄疸等临床症状的改变进行判断。适用于国外较常见的造血干细胞移植后肝窦阻塞综合征(HSCT-HSOS)。而我国以PA-HSOS为主,其中因服用土三七导致的HSOS占50.0%~88.6%,肝组织病理学检查、血管造影等是PA-HSOS的重要方法,但是是创伤性的,加上大量患者不能准确回忆服药史,也无法提供服用的可疑药物或食物,导致即使肝脏病理检查发现肝窦淤血等组织学改变,也不能获得确切病因诊断。相当一部分患者没有得到及时诊断和恰当的治疗,导致预后不佳。
发展有效的精准诊断方法有助于解决PAs导致的HSOS诊治的瓶颈问题。针对PAs的致病机制,科学家将血浆等含毒素生物样本经生化处理后,将上清液与AgNO3、乙醇共孵育,将吡咯里西啶生物碱加成分子群体归一转化为关键的二羟乙基脱氢吡咯里西啶生物碱。
随后经与DABA进行衍生化反应原位生成二甲氨苄吡咯里西啶碱(DABP),用于HPLC-MS或UPLC-MS色质联用检测。经过与化学合成的DABP标准品进行HPLC-MS或UPLC-MS色质联用技术检测比对,通过检测相应分子的保留时间,可以达到得到病人来源样品的PAs存在的确切依据。通过对病人来源样品的PAs存在丰度进一步进行定量研究,还可以得到毒素含量与肝损伤程度的定量关系。
HSOS生物标志物检测过程中,获取二羟乙基脱氢吡咯里西啶生物碱标准品是建立HSOS疾病标志物检测方案的关键因素。此前,有相关文献(Chem.Biol. Interact.1990,75,225-239;Tetrahedron,2002,58:3553-3559;Chem.Res.Toxicol. 2016,29:1282-1292)报道了该分子的合成过程。如Chem.Biol.Interact.1990,75, 225-239报道合成了二羟乙基脱氢吡咯里西啶生物碱,但是文献中没有提供任何收率与相关化合物核磁图谱。Tetrahedron,2002,58:3553-3559文献报道,仅能合成毫克级别脱氢野百合碱,严重限制了二羟乙基脱氢吡咯里西啶生物碱的规模化合成。而按照Chem.Res.Toxicol.2016,29:1282-1292文献报道,也仅能以65.7%收率从脱氢野百合碱合成得到毫克级别二羟乙基脱氢吡咯里西啶生物碱。
这些文献合成方法都存在合成规模较小(毫克级别)、反应选择性低副产物多、合成方法复杂、产品收率非常低、难以实现放大合成等问题,远远无法满足现代大规模临床诊断的需要。因此,大幅度提高该关键分子的合成规模与效率,对于药物性肝损伤疾病的精准医疗体系的建立至关重要。
发明内容
本发明的目的是:提供用于一类中草药导致的肝损伤疾病诊断所需重要生物标志物二羟乙基脱氢吡咯里西啶生物碱的新合成工艺,大幅度提高了该关键分子的合成收率,达到生产级别要求,为该类中草药导致的肝损伤疾病的精准医疗体系的建立提供关键技术支撑。
为了实现上述目的,本发明提供了一类中草药肝损伤相关生物标志物的化学合成方法,所述的生物标志物为二羟乙基脱氢吡咯里西啶生物碱,其化学合成方法具体包括如下步骤:
步骤1:以天然吡咯里西啶生物碱为原料,在有机溶剂中溶解后,加入氧化剂和催化剂进行氧化反应,然后加入稀酸回流反应,反应结束后,静置分层,收集有机相并用稀碱液进行洗涤至水相洗液呈中性,然后干燥,在干燥的有机相中冰浴下滴加金属醇盐溶液,然后升温反应,反应完成后,稀酸淬灭反应,过滤除去沉淀,收集有机相,依次经过洗涤、干燥和浓缩后得到产物脱氢吡咯里西啶生物碱;
步骤2:将步骤1所得的吡咯里西啶生物碱溶解后和路易斯酸或路易斯酸的醚溶液反应得到二羟乙基脱氢吡咯里西啶生物碱或其醚化产物;
所述步骤1中的天然吡咯里西啶生物碱为下列式I所示的7-位单酯取代类吡咯里西啶生物碱、式II所示的9-位单酯取代类吡咯里西啶生物碱、式III所示的 7,9-二酯取代开环类吡咯里西啶生物碱、或式IV所示的11元大环双酯类吡咯里西啶生物碱或12元大环双酯类吡咯里西啶生物碱:
相应地,所述步骤1所得的脱氢吡咯里西啶生物碱的结构式如Ia-IVa所示:
优选地,所述结构式中的R1、R2分别独立地选自C1-C4的烷烃,Rx为丙撑基或丁撑基。更优选地,所述天然吡咯里西啶生物碱为野百合碱,野百合碱为常规市售的商业化产品。
优选地,所用步骤1中的氧化剂选自四氯苯醌,史试剂Shi Reagen和二甲亚砜(DMSO)-草酰氯(Swern试剂)中的至少一种;所述氧化剂的用量为天然吡咯里西啶生物碱的摩尔量的1~3倍。其中更优选的氧化试剂为四氯苯醌。
优选地,所述步骤1中的催化剂为过渡金属类催化剂,所述催化剂的用量为天然吡咯里西啶生物碱的摩尔量的5~20%,所述有机溶剂为氯代烷烃。
优选地,所述过渡金属类催化剂为三氟醋酸钯,所述氯代烷烃选自二氯甲烷和/或氯仿。
优选地,所述步骤1中的金属醇盐选自甲醇钠、乙醇钠和叔丁醇钾中的至少一种。
优选地,所述步骤2中的路易斯酸选自三氟化硼、三氟化硼乙醚、三氯化铝、三氧化硫和二氯卡宾中的至少一种,所述路易斯酸的用量为脱氢吡咯里西啶生物碱的摩尔量的0.6~2倍。其中更优选的Lewis酸为三氟化硼乙醚。
优选地,所述步骤2中的有机溶剂为醇溶剂。
与现有技术相比,本发明的有益效果在于:
(1)本发明的生物标志物二羟乙基脱氢吡咯里西啶生物碱的合成方法,以可商购的天然吡咯里西啶生物碱为起始原料,放大反应后反应选择性好、副反应少,且产物易于纯化,本发明的制备过程简单、收率高、易操作,且适合放大合成及工业化生产;
(2)本发明的合成方法大幅度提高了该生物标志物的合成收率,达到生产级别要求,为吡咯里西啶生物碱类中草药导致的肝损伤疾病的精准医疗体系的建立提供关键技术支撑。
具体实施方式
为使本发明更明显易懂,兹以优选实施例,作详细说明如下。
实施例1:脱氢吡咯里西啶生物碱的制备
将四氯苯醌(113克,461mmol)和催化剂三氟乙酸钯(30mmol)溶解在500mL 二氯甲烷中搅拌均匀。缓慢滴入500mL野百合碱(100克,307mmol)的二氯甲烷溶液,并将溶液快速搅拌1小时。随后加入稀盐酸溶液(1M,200mL),回流下剧烈搅拌24h,冷却至室温后,收集有机相,加入稀NaOH溶液水洗至水相洗液呈中性,并干燥有机相,冰浴下,在干燥的有机相中缓慢滴加叔丁醇钾的四氢呋喃溶液(12M,100mL),滴加完毕后,升温至回流下继续搅拌反应4h,反应在无水无氧条件下进行。反应结束后,加稀盐酸淬灭反应,过滤掉固体沉淀物,用盐水洗涤有机层2~3次并用Na2SO4干燥。过滤并浓缩,得到89g脱氢野百合碱化合物,收率90%。1H NMR(300MHz,DMSO-d6):δ6.67(d,J=2.5Hz,1H), 6.19(d,J=2.5Hz,1H),5.97(dd,J=3.0,8Hz,1H),5.16(s,1H),5.11(d,J=12Hz, 1H),4.41(d,J=12Hz,1H),4.37(s,1H),4.11(m,1H),3.95(m,1H),3.03(m,1H), 2.93(m,1H),2.44(m,1H),1.35(s,3H),1.25(s,3H),1.14(d,J=7Hz,3H);ESI-MS: 324.1[M+1]。
实施例2:二羟乙基脱氢吡咯里西啶生物碱的制备
将脱氢野百合碱(50g,155mmol)溶于1升乙醇中,零度下滴入三氟化硼乙醚溶液100mL。在室温下搅拌18小时后停止反应。减压浓缩溶液。粗产物通过硅胶柱层析纯化,得到27.5g淡黄色油状化合物二羟乙基脱氢吡咯里西啶生物碱收率,85%。1H NMR(500MHz,Acetone-d6):δ6.63(d,J=2.7Hz,1H),6.12(d, J=2.7Hz,1H),4.82(dd,J=6.1,1H),4.41(d,J=11.4Hz,1H),4.35(d,J=11.4Hz, 1H),4.07(m,1H),3.93(m,1H),3.52(m,2H),3.48(m,2H),2.71(m,1H),2.46(m, 1H),1.16(t,J=7Hz,3H),1.15(t,J=7Hz,3H);13C NMR(126MHz,acetone)δ 206.14,115.02,113.48,74.22,66.20,65.08,63.75,45.17,37.14,29.84,15.67; ESI-MS:210.1[M+1]。
对比例
采用文献报道方法合成脱氢吡咯里西啶生物碱:
将四氯苯醌(113克,461毫摩尔)溶解在500mL二氯甲烷中搅拌均匀。缓慢滴入500mL野百合碱(100克,307毫摩尔)的二氯甲烷溶液,并将溶液快速搅拌1小时。随后分批加入30克NaBH4,剧烈搅拌90分钟。然后再快速滴入 500mL10%NaOH水溶液。继续搅拌1小时后停止反应。过滤掉绿色沉淀物,用盐水洗涤有机层并用Na2SO4干燥。过滤并浓缩,得到混合物,经过柱层析和液氮下重结晶未得到目标产物组分。
上述实施例仅为本发明的优选实施例,并非对本发明任何形式上和实质上的限制,应当指出,对于本技术领域的普通技术人员,在不脱离本发明的前提下,还将可以做出若干改进和补充,这些改进和补充也应视为本发明的保护范围。
Claims (7)
1.一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所述的生物标志物为二羟乙基脱氢吡咯里西啶生物碱,其化学合成方法具体包括如下步骤:
步骤1:以天然吡咯里西啶生物碱为原料,在有机溶剂中溶解后,加入氧化剂和催化剂三氟醋酸钯进行氧化反应,然后加入稀酸回流反应,反应结束后,静置分层,收集有机相并用稀碱液进行洗涤至水相洗液呈中性,然后干燥,在干燥的有机相中冰浴下滴加过量的金属醇盐溶液,然后升温反应,反应完成后,稀酸淬灭反应,过滤除去沉淀,收集有机相,依次经过洗涤、干燥和浓缩后得到产物脱氢吡咯里西啶生物碱;
步骤2:将步骤1所得的吡咯里西啶生物碱溶解后和路易斯酸或路易斯酸的醚溶液反应得到二羟乙基脱氢吡咯里西啶生物碱或其醚化产物;
所述步骤1中的天然吡咯里西啶生物碱为下列式I所示的7-位单酯取代类吡咯里西啶生物碱、式II所示的9-位单酯取代类吡咯里西啶生物碱、式III所示的7,9-二酯取代开环类吡咯里西啶生物碱、或式IV所示的11元大环双酯类吡咯里西啶生物碱或12元大环双酯类吡咯里西啶生物碱:
、/>、/>、/>;
相应地,所述步骤1所得的脱氢吡咯里西啶生物碱的结构式如Ia-IVa所示:
、/>、/>、/>。
所述结构式中的R1、R2分别独立地选自C1-C4的烷烃,Rx为丙撑基或丁撑基。
2.如权利要求1所述的一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所用步骤1中的氧化剂选自四氯苯醌,史试剂Shi Reagent和二甲亚砜-草酰氯中的至少一种;所述氧化剂的用量为天然吡咯里西啶生物碱的摩尔量的1~3倍。
3.如权利要求1所述的一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所述步骤1中催化剂的用量为天然吡咯里西啶生物碱的摩尔量的5~20%,所述有机溶剂为氯代烷烃。
4.如权利要求3所述的一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所述氯代烷烃选自二氯甲烷和/或氯仿。
5.如权利要求1所述的一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所述步骤1中的金属醇盐选自甲醇钠、乙醇钠和叔丁醇钾中的至少一种。
6.如权利要求1所述的一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所述步骤2中的路易斯酸选自三氟化硼、三氟化硼乙醚、三氯化铝、三氧化硫和二氯卡宾中的至少一种,所述路易斯酸的用量为脱氢吡咯里西啶生物碱的摩尔量的0.6~2倍。
7.如权利要求1所述的一类中草药肝损伤相关生物标志物的放大合成方法,其特征在于,所述步骤2中的有机溶剂为醇溶剂。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010099740A1 (zh) * | 2009-03-03 | 2010-09-10 | 中国医学科学院药物研究所 | 13a-(S)去氧娃儿藤宁衍生物、其制法和药物组合物与用途 |
WO2017050262A1 (zh) * | 2015-09-25 | 2017-03-30 | 南开大学 | 菲并吲哚里西啶生物碱季铵盐衍生物及其制备和抗植物病毒应用 |
EP3159002A1 (de) * | 2015-10-22 | 2017-04-26 | Merck Patent GmbH | Verfahren zur herstellung von spezial-extrakten aus symphytum |
EP3412299A1 (de) * | 2017-06-09 | 2018-12-12 | SMC - Research AG | Eintopf-verfahren bei der herstellung von spezialextrakten aus symphytum |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010099740A1 (zh) * | 2009-03-03 | 2010-09-10 | 中国医学科学院药物研究所 | 13a-(S)去氧娃儿藤宁衍生物、其制法和药物组合物与用途 |
WO2017050262A1 (zh) * | 2015-09-25 | 2017-03-30 | 南开大学 | 菲并吲哚里西啶生物碱季铵盐衍生物及其制备和抗植物病毒应用 |
EP3159002A1 (de) * | 2015-10-22 | 2017-04-26 | Merck Patent GmbH | Verfahren zur herstellung von spezial-extrakten aus symphytum |
EP3412299A1 (de) * | 2017-06-09 | 2018-12-12 | SMC - Research AG | Eintopf-verfahren bei der herstellung von spezialextrakten aus symphytum |
Non-Patent Citations (2)
Title |
---|
Jetze J. Tepe.DNA interstrand cross-link formation by reductive activation of dehydropyrrolizidine progenitors.Tetrahedron.2002,第58卷第3553页Scheme 1,第3556页. * |
Yang, Xiaojing et al..A Difference in Internal Exposure Makes Newly Weaned Mice More Susceptible to the Hepatotoxicity of Retrorsine Than Adult Mice.Chemical Research in Toxicology.2018,第31卷(第12期),第1349页Scheme 1. * |
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