CN117486962A - 一种人参皂苷衍生物及其制备方法和应用 - Google Patents
一种人参皂苷衍生物及其制备方法和应用 Download PDFInfo
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- CN117486962A CN117486962A CN202311431547.2A CN202311431547A CN117486962A CN 117486962 A CN117486962 A CN 117486962A CN 202311431547 A CN202311431547 A CN 202311431547A CN 117486962 A CN117486962 A CN 117486962A
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- ginsenoside
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Abstract
本发明涉及一种人参皂苷衍生物及其制备方法和应用。人参皂苷衍生物,其结构为如式(I)所述的化合物及其立体异构体或其药学上可接受的盐。本发明还提供人参皂苷衍生物的制备方法。本发明还提供人参皂苷衍生物在制备疫苗中的应用,该人参皂苷衍生物具有原人参二醇或原人参三醇类似物及其骨架,且对动物模型,表现出较高的佐剂活性,解决了现有疫苗佐剂匮乏等问题。
Description
技术领域
本发明涉及药物化学技术领域,具体涉及一种人参皂苷衍生物及其制备方法和应用。
背景技术
疫苗免疫是预防各种传染病最有效和经济的方式。随着分子生物学和基因工程技术的发展,类毒素疫苗和亚单位疫苗应运而生。一般而言,减毒活疫苗本身带有免疫刺激信号,足以引起获得性免疫应答。而类毒素疫苗和亚单位疫苗的抗原可能缺乏启动免疫应答的某些免疫信息以及消耗过快等因素,往往需要一些佐剂来增强或调节免疫原性。目前,被发现有佐剂活性的物质多种多样。按作用机理分,佐剂大致可以分为2类:免疫增强剂类(Toll样受体激动剂、皂苷、细胞因子等)和抗原递送剂类(矿物盐、乳剂、微粒等)。按化学性质分,佐剂可以分为5类:矿物盐、油乳剂、靶向模式识别受体的化合物、细胞因子和天然佐剂。
其中,矿物盐佐剂主要是铝盐佐剂。铝盐佐剂目前是动物疫苗中应用最广泛的佐剂。但是铝盐佐剂引发的细胞坏死和炎性激活会造成注射位点疼痛等轻微毒性反应。同时,铝盐佐剂还会诱导产生IL-1,因此,易导致头疼、关节疼和肌肉疼等症状。铝盐佐剂还存在另一个潜在的问题,它诱导Th2型免疫反应的偏好性,增加了嗜酸性粒细胞和IgE抗体反应,从而增加过敏反应的风险。油乳佐剂通过油乳本身对抗原的包裹使抗原可以在机体内持续存在,从而使抗原刺激的时间延长,免疫效果提高。但也正是因为油乳佐剂这个特点,在持续的刺激下,机体中容易出现组织损伤、应激反应、矿物油的残留等,从而存在一定的安全隐患。细胞因子佐剂在使用的过程中虽然可以促进特异性免疫反应,可以激发非特异性免疫。但是就临床应用而言,目前细胞因子佐剂还存在半衰期短、价格昂贵等问题。除此之外,对剂量的把握也是细胞因子佐剂在应用过程中亟待解决的难题,因为剂量使用不当,很容易导致潜在的自身免疫安全隐患。壳聚糖佐剂由于其水溶性差和黏膜免疫障碍,目前尚未在临床上得到广泛应用。皂苷类免疫激活剂QS-21同样面临诸多应用问题:①QS-21是从植物中提取的,存在含量低、成本高的难题;②QS-21的稳定性较差,在生理条件下会被水解;③QS-21的毒性虽然相对于QuilA有所减弱,但是在高剂量下,仍有较大毒性并伴有疼痛感。因此,研发安全有效、性质稳定的佐剂迫在眉睫。
发明内容
本发明的目的在于提供一种人参皂苷衍生物及其制备方法和应用,以解决背景技术中的佐剂存在的至少一个问题。
为了实现上述目的,本发明采用的技术方案如下:
一种人参皂苷衍生物,其结构为如式(I)所述的化合物及其立体异构体或其药学上可接受的盐:
式(I)中,
R1选自氢或糖基;
R2选自氢、羧基、酯、酰胺、杂脂肪基、烷氧基脂肪基、芳基氧脂肪基或环状基;
R3选自氢或-ORy,所述Ry为氢或糖基;
n的取值范围为0~10。
本发明提供的人参皂苷衍生物中具有原人参二醇或原人参三醇类似物及其骨架,经过实验证明,将本发明的人参皂苷衍生物作为疫苗佐剂,表现出了较高的佐剂活性。
优选的,所述糖基选自:β-D-葡萄糖基、α-D-葡萄糖基、β-D-半乳糖基、α-D-半乳糖基、β-D-甘露糖基、α-D-甘露糖基、β-D-木糖基、α-D-木糖基、β-D-2-氨基葡萄糖基、α-D-2-氨基葡萄糖基、α-L-鼠李糖基、β-L-鼠李糖基、α-D-阿拉伯糖基、β-D-阿拉伯糖基、α-L-阿拉伯糖基、β-L-阿拉伯糖基、α-L-岩藻糖基、β-L-岩藻糖基、β-D-葡萄糖醛酸基、α-D-葡萄糖醛酸基、β-D-半乳糖醛酸基、α-D-半乳糖醛酸基,或者为上述单糖基组成的2~6个寡糖糖链基。
优选的,所述芳基氧脂肪基或环状基存在0~5个取代基Rx,所述取代基Rx选自芳基、卤代芳基、环烷基芳基、卤代烷基芳基、卤代烷氧基芳基、杂环基、卤代杂环基、杂芳基、卤代杂芳基、卤代烷基杂芳基、卤代烷氧基杂芳基、烷基甲酰基、芳基甲酰基、卤代芳基甲酰基、杂芳基甲酰基、卤代杂芳基甲酰基、卤代芳基磺酰基、羟基烷基、羟基炔基、氨基、氨基烷基、氢、卤素、羧基、酯基、氰基、C1-4烷基或C1-4烷氧基。
优选的,所述n的取值范围为0~10之间的整数。
优选的,所述环状基选自芳基、杂芳基、环烷基或杂环烷基。
优选的,R2选自 其中,X选自O、NH或S,Z选自H、烷基、芳基或杂芳基,n的取值范围为0~10。
优选的,式(I)所述的化合物为
本发明还提供一种人参皂苷衍生物的制备方法,包括以下步骤:
式(II)所示的化合物与式(V-1)所示的糖给体或式(V-2)所示的糖给体反应,得到式(III)所示的化合物;
式(III)所示的化合物与式(VI-1)所示的葡萄糖衍生物或式(VI-2)所示的葡萄糖衍生物反应,得到式(IV)所示的化合物;
式(IV)所示的化合物脱烯丙基和乙酰基,得到式(I)所示的化合物。
根据上述技术手段,通过苷元与糖基供体反应合成人参皂苷衍生物,合成工艺具有条件简单温和、生产成本较低的优点,适合开发成疫苗佐剂,在药物化学技术领域,具有潜在的新药开发价值。
优选的,包括以下步骤:
式(II)所示的化合物溶于含有路易斯酸的有机溶剂中,加入式(V-1)所示的糖给体或式(V-2)所示的糖给体反应,纯化得到式(III)所示的化合物;
式(III)所示的化合物溶于有机溶剂,加入脱硅基保护基试剂,搅拌反应液,纯化得到第一中间产物;将第一中间产物溶于含有路易斯酸的有机溶剂中,加入式(VI-1)所示的葡萄糖衍生物或式(VI-2)所示的葡萄糖衍生物,搅拌反应,纯化得到式(IV)所示的化合物;
式(IV)所示的化合物溶于有机溶剂中,加入脱烯丙基金属催化剂,搅拌反应液,纯化得到第二中间产物;将第二中间产物溶于有机溶剂中,在碱的作用下脱去乙酰基,纯化得到式(I)所示的化合物。
优选的,所述式(II)所示的化合物、式(V-1)所示的糖给体或式(V-2)所示的糖给体与路易斯酸的摩尔比为1:1.2~5.0:0.05~0.5。
优选的,所述式(III)所示的化合物、式(VI-1)所示的葡萄糖衍生物或式(VI-2)所示的葡萄糖衍生物与路易斯酸的摩尔比为1:2.0~5.0:0.05~0.5。
优选的,所述有机溶剂选自吡啶、氯仿、二氯甲烷、乙酸乙酯、四氢呋喃、1,4-二氧六环、氯代苯、甲苯、乙腈、N,N-二甲基甲酰胺或二甲基亚砜。
优选的,所述脱硅基保护基试剂选自四甲基氟化铵、四乙基氟化铵、四丁基氟化铵(TBAF)或醋酸。
优选的,所述脱烯丙基金属催化剂选自氯化钯(PdCl2)、醋酸钯(Pd(OAc)2)或四三苯基膦钯(Pd(PPh3)4);
优选的,所述路易斯酸选自分子筛、三氟化硼乙醚(BF3·Et2O)、三氟甲磺酸三甲基硅脂(TMSOTf)、氯化金(AuCl)、三氯化金(AuCl3)、三苯基膦金(I)三氟甲烷磺酸盐(Ph3PAuOTf)或三苯基膦双(三氟甲磺酰亚胺)金(I)(Ph3PAuNTf2),所述分子筛选自分子筛或酸洗分子筛,底物与分子筛的质量比为1:2.0~5.0;
优选的,所述碱为甲醇钠(MeONa)、氢氧化钠(NaOH)或氢氧化钾(KaOH)。
优选的,所述式(II)所示的化合物与式(V-1)所示的糖给体或式(V-2)所示的糖给体的反应在低温或室温条件下进行,所述低温的温度在-78℃~0℃之间。
优选的,其特征在于,所述式(IV)所示化合物为:
本发明还提供一种人参皂苷衍生物的应用,所述人参皂苷衍生物在制备疫苗中的应用。
经过实验证明,本发明提供的人参皂苷衍生物对动物模型,表现出较高的佐剂活性,从而证明了人参皂苷衍生物在疫苗佐剂中具有潜在的应用前景,很好地解决了目前疫苗佐剂匮乏的问题。
优选的,式(I)所述的化合物及其立体异构体或其药学上可接受的盐在制备疫苗中作为佐剂的应用。
本发明的有益效果:
本发明提供的人参皂苷衍生物中具有原人参二醇或原人参三醇类似物及其骨架。人参皂苷类化合物在免疫调节、抗肿瘤和治疗神经系统疾病方面展现出巨大的潜力。比如,原人参三醇型人参皂苷Rg1可以选择性地增加老龄鼠的脾脏淋巴细胞的增殖能力,并且显著促进IL-2的产生与释放。弓形虫重组蛋白ROP18与原人参三醇型人参皂苷Re联合应用对小鼠免疫效果研究表明,添加人参皂苷Re可使ROP18诱导产生更强的体液和细胞免疫反应皂苷。将金黄色葡萄球菌抗原单独注射豚鼠时,产生的特异性抗体的含量很低。而原人参二醇型皂苷Rb1作为免疫佐剂显著提高了其相应的特异性抗体水平。将Rb1作为免疫佐剂与乳房炎金黄色葡萄球菌疫苗同时注射奶牛后,血清检验发现Th1型与Th2型的细胞因子的含量均发生了变化。实验证明,将本发明的人参皂苷衍生物作为疫苗佐剂,表现出了较高的佐剂活性;
本发明提供的人参皂苷衍生物的制备方法,首先通过皂苷苷元与糖基供体反应,然后在室温下先后脱掉羟基上的硅基保护基、烯丙基保护基和乙酰基保护基,合成得到人参皂苷衍生物。目标产物合成过程中所用试剂和溶剂均为常规试剂且多数反应均在室温下就能完成。因此合成工艺具有条件简单温和、生产成本较低的优点,适合开发成疫苗佐剂,在药物化学技术领域,具有潜在的新药开发价值;
本发明提供的人参皂苷衍生物对动物模型,表现出较高的佐剂活性,从而证明了人参皂苷衍生物在疫苗佐剂中具有潜在的应用前景,很好地解决了目前疫苗佐剂匮乏的问题。
附图说明
图1为血清特异性抗体IgG的检测结果图;
图2为血清特异性抗体亚类IgG1的检测结果图;
图3为血清特异性抗体亚类IgG2a的检测结果图。
具体实施方式
以下将参照附图和优选实施例来说明本发明的实施方式,本领域技术人员可由本说明书中所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用,本说明书中的各项细节也可以基于不同观点与应用,在没有背离本发明的精神下进行各种修饰或改变。应当理解,优选实施例仅为了说明本发明,而不是为了限制本发明的保护范围。
实施例1
一种如化合物1所示的人参皂苷衍生物的制备方法,
其中,化合物1的合成过程如下:
制备方法的步骤包括:
步骤1:将C-3和C-12位羟基保护的原人参二醇衍生物1-1(300.0mg,0.43mmol)和糖给体1a(453.0mg,0.52mmol)置于含有干燥二氯甲烷(20mL)的烧瓶中,抽真空置换氩气;在-78℃和氩气保护条件下,加入三氟化硼乙醚溶液(65μL,0.52mmol)进行反应;反应结束后,在-78℃条件下加入三乙胺淬灭反应;室温条件下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得目标化合物1-2(产量:341.2mg,收率:56%),HRMS(MALDI)C76H124O22SiNa[M+Na]+预估值:1439.8251,实测值:1439.8239。
步骤2:将目标化合物1-2(280.0mg,0.20mmol)加入含有干燥四氢呋喃(12mL)的烧瓶中,室温条件下加入1mol/L的四丁基氟化胺的四氢呋喃溶液(2mL,2.0mmol)和冰醋酸(150μL),室温搅拌12小时;反应结束后,用乙酸乙酯稀释,然后水洗(20mL×3),无水硫酸钠干燥,过滤,室温下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得中间体I-1(产量:202.4mg,收率:83%);将中间体I-1(160.0mg,0.13mmol)和化合物1b(112.0mg,0.16mmol)加入含有无水二氯甲烷(12mL)的烧瓶中,抽真空置换氩气;在-78℃和氩气保护条件下,加入三氟化硼乙醚溶液(20μL,0.16mmol)。反应结束后,-78℃条件下加入三乙胺淬灭反应;室温条件下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得目标化合物1-3(产量:108.0mg,收率:47%),HRMS(MALDI)C92H143NO32Na[M+Na]+预估值:1773.9593,实测值:1773.9576。
步骤3:将目标化合物1-3(70.0mg,0.04mmol)溶于含有甲醇/二氯甲烷(7mL/7mL)的烧瓶中,加入PdCl2(7.0mg),室温条件下搅拌过夜;反应结束后,用硅藻土过滤,室温下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得中间体I-2(产量:54.6mg,收率:77%);将中间体I-2(40.0mg,0.02mmol)加入含有10%氢氧化钾/甲醇溶液(0.4g:5mL)的烧瓶中,室温搅拌12小时,反应结束后,加入酸性树脂淬灭反应,调节pH至7~8,过滤树脂,室温条件下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,二氯甲烷/甲醇(体积比:15/1至8/1)作洗脱剂,得终产物1(产量:18.0mg,收率:71%),即人参皂苷衍生物,HRMS(MALDI)C65H111NO22Na[M+Na]+预估值:1280.7495,实测值:1280.7472。
实施例2
一种如化合物2所示的人参皂苷衍生物的制备方法,
其中,化合物2的合成过程如下:
制备方法的步骤包括:
步骤1:将C-3和C-12位羟基保护的原人参二醇衍生物2-1(400.0mg,0.65mmol)和糖给体1a(690.0mg,0.78mmol)置于含有干燥二氯甲烷(20mL)的烧瓶中,抽真空置换氩气;在-78℃和氩气保护条件下,加入三氟化硼乙醚溶液(80μL,0.78mmol);反应结束后,在-78℃的条件下加入三乙胺淬灭反应;室温下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得目标化合物2-2(产量:431.1mg,收率:45%),HRMS(MALDI)C76H124O22SiNa[M+Na]+预估值:1497.8306,实测值:1497.8283;
步骤2:将目标化合物2-2(360.0mg,0.24mmol)加入含有干燥四氢呋喃(18mL)的烧瓶中,室温条件下加入含有四丁基氟化胺的四氢呋喃溶液(2.4mL,2.4mmol)和冰醋酸(200μL),室温搅拌12小时;反应结束后,用乙酸乙酯稀释,然后水洗(30mL×3),无水硫酸钠干燥,过滤,室温条件下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得中间体II-1(产量:263.6mg,收率:86%);将中间体II-1(200.0mg,0.16mmol)和化合物1b(136.0mg,0.19mmol)加入含有无水二氯甲烷(15mL)的烧瓶中,抽真空置换氩气;在-78℃和氩气保护条件下,加入三氟化硼乙醚溶液(23μL,0.19mmol);反应结束后,-78℃条件下加入三乙胺淬灭反应;室温条件下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至1/1)为洗脱剂,得目标化合物2-3(产量:111.4mg,收率:38%),HRMS(MALDI)C94H145NO34Na[M+Na]+预估值:1854.9546,实测值:1854.9522;
步骤3:将目标化合物2-3(80.0mg,0.04mmol)溶于含有甲醇/二氯甲烷(7mL/7mL)的烧瓶中,加入二氯化钯(10.0mg),室温下搅拌过夜;反应结束后,用硅藻土过滤,室温下减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,石油醚和乙酸乙酯(体积比:5/1至2/1)为洗脱剂,得中间体II-2(产量:58.7mg,收率:82%);将中间体II-2(45.0mg,0.025mmol)加入含有10%氢氧化钾/甲醇溶液(0.4g:5mL)的烧瓶中,室温搅拌12小时,反应结束后,加入酸性树脂淬灭反应,调节pH至7~8,过滤树脂,室温减压浓缩溶剂,粗产物用硅胶柱层析进行纯化,二氯甲烷/甲醇(体积比:12/1至8/1)作洗脱剂,得终产物2(产量:24.0mg,收率:76%),即人参皂苷衍生物,HRMS(MALDI)C65H111NO23Na[M+Na]+预估值:1296.7445,实测值:1296.7416。
检测分析
测试实施例1中制得的终产物1和实施例2中制得的终产物2对免疫模式抗原(卵清白蛋白)(OVA)小鼠的免疫佐剂作用。
具体操作为:将60只小鼠随机分为5组,分别为:无免疫空白组(CK)、免疫阴性组(OVA)、免疫阳性组(氢氧化铝,100μg)(AL)、免疫实验组1(化合物1,100μg)(Exp 1)和免疫实验组2(化合物2,100μg)(Exp 2);
适应性饲养7天后,进行第一次免疫,分别对无免疫空白组(CK)的小鼠腹股沟皮下注射无菌PBS 0.2mL,对免疫阴性组(OVA)的小鼠腹股沟皮下注射OVA(0.2mL,100μg),对免疫阳性组(AL)的小鼠腹股沟皮下注射氢氧化铝(100μg),对免疫实验组1(Exp 1)的小鼠腹股沟皮下注射化合物1(100μg),对免疫实验组2(Exp 2)的小鼠腹股沟皮下注射化合物2(100μg);
第21天进行第二次免疫,分别对无免疫空白组(CK)的小鼠腹股沟皮下注射无菌PBS 0.2mL,对免疫阴性组(OVA)的小鼠腹股沟皮下注射OVA(0.2mL,100μg),对免疫阳性组(AL)的小鼠腹股沟皮下注射氢氧化铝(100μg),对免疫实验组1(Exp 1)的小鼠腹股沟皮下注射化合物1(100μg),对免疫实验组2(Exp 2)的小鼠腹股沟皮下注射化合物2(100μg);
第二次免疫后,第14天取眼静脉,解剖处死小鼠,采用ELISA法测定用OVA免疫的小鼠血清中免疫球蛋白IgG、IgG1和IgG2a的浓度,结果分别如图1至图3所示。
从图1中分析可知,化合物1和化合物2可大幅度提高OVA抗原诱导的IgG水平,且优于佐剂氢氧化铝胶。
从图2中分析可知,化合物1和化合物2以及氢氧化铝胶均能提高IgG1的浓度,这表明化合物1和化合物2可以有效促进体液免疫反应。
从图3中分析可知,化合物1和化合物2可以显著提高IgG2的浓度,而氢氧化铝则对IgG2没有显著的促进作用,这表明化合物1和化合物2能促进细胞免疫反应。
上述测试结果证明了化合物1和化合物2在作为疫苗佐剂方面具有潜在的开发价值。
综上所述,本发明提供的人参皂苷衍生物中具有原人参二醇或原人参三醇类似物及其骨架,经过实验证明,将本发明的人参皂苷衍生物作为疫苗佐剂,表现出了较高的佐剂活性。
本发明提供的人参皂苷衍生物的制备方法,通过苷元与糖基供体反应合成人参皂苷衍生物,合成工艺具有条件简单温和、生产成本较低的优点,适合开发成疫苗佐剂,在药物化学技术领域,具有潜在的新药开发价值。
本发明提供的人参皂苷衍生物对动物模型,表现出较高的佐剂活性,从而证明了人参皂苷衍生物在疫苗佐剂中具有潜在的应用前景,很好地解决了目前疫苗佐剂匮乏的问题。
以上实施例仅是为充分说明本发明而所举的较佳的实施例,本发明的保护范围不限于此。本技术领域的技术人员在本发明基础上所作的等同替代或变换,均在本发明的保护范围之内。
Claims (10)
1.一种人参皂苷衍生物,其特征在于,其结构为如式(I)所述的化合物及其立体异构体或其药学上可接受的盐:
式(I)中,
R1选自氢或糖基;
R2选自氢、羧基、酯、酰胺、杂脂肪基、烷氧基脂肪基、芳基氧脂肪基或环状基;
R3选自氢或-ORy,所述Ry为氢或糖基;
n的取值范围为0~10。
2.根据权利要求1所述的人参皂苷衍生物,其特征在于,所述糖基选自:β-D-葡萄糖基、α-D-葡萄糖基、β-D-半乳糖基、α-D-半乳糖基、β-D-甘露糖基、α-D-甘露糖基、β-D-木糖基、α-D-木糖基、β-D-2-氨基葡萄糖基、α-D-2-氨基葡萄糖基、α-L-鼠李糖基、β-L-鼠李糖基、α-D-阿拉伯糖基、β-D-阿拉伯糖基、α-L-阿拉伯糖基、β-L-阿拉伯糖基、α-L-岩藻糖基、β-L-岩藻糖基、β-D-葡萄糖醛酸基、α-D-葡萄糖醛酸基、β-D-半乳糖醛酸基、α-D-半乳糖醛酸基,或者为上述单糖基组成的2~6个寡糖糖链基;
和/或所述芳基氧脂肪基或环状基存在0~5个取代基Rx,所述取代基Rx选自芳基、卤代芳基、环烷基芳基、卤代烷基芳基、卤代烷氧基芳基、杂环基、卤代杂环基、杂芳基、卤代杂芳基、卤代烷基杂芳基、卤代烷氧基杂芳基、烷基甲酰基、芳基甲酰基、卤代芳基甲酰基、杂芳基甲酰基、卤代杂芳基甲酰基、卤代芳基磺酰基、羟基烷基、羟基炔基、氨基、氨基烷基、氢、卤素、羧基、酯基、氰基、C1-4烷基或C1-4烷氧基;
和/或所述n的取值范围为0~10之间的整数;
和/或所述环状基选自芳基、杂芳基、环烷基或杂环烷基。
3.根据权利要求1所述的人参皂苷衍生物,其特征在于,R2选自
其中,X选自O、NH或S,Z选自H、烷基、芳基或杂芳基,n的取值范围为0~10。
4.根据权利要求1所述的人参皂苷衍生物,其特征在于,式(I)所述的化合物为
5.一种如权利要求1至权利要求4任一项所述的人参皂苷衍生物的制备方法,其特征在于,包括以下步骤:
式(II)所示的化合物与式(V-1)所示的糖给体或式(V-2)所示的糖给体反应,得到式(III)所示的化合物;
式(III)所示的化合物与式(VI-1)所示的葡萄糖衍生物或式(VI-2)所示的葡萄糖衍生物反应,得到式(IV)所示的化合物;
式(IV)所示的化合物脱烯丙基和乙酰基,得到式(I)所示的化合物。
6.根据权利要求5所述的人参皂苷衍生物的制备方法,其特征在于,包括以下步骤:
式(II)所示的化合物溶于含有路易斯酸的有机溶剂中,加入式(V-1)所示的糖给体或式(V-2)所示的糖给体反应,纯化得到式(III)所示的化合物;
式(III)所示的化合物溶于有机溶剂,加入脱硅基保护基试剂,搅拌反应液,纯化得到第一中间产物;将第一中间产物溶于含有路易斯酸的有机溶剂中,加入式(VI-1)所示的葡萄糖衍生物或式(VI-2)所示的葡萄糖衍生物,搅拌反应,纯化得到式(IV)所示的化合物;
式(IV)所示的化合物溶于有机溶剂中,加入脱烯丙基金属催化剂,搅拌反应液,纯化得到第二中间产物;将第二中间产物溶于有机溶剂中,在碱的作用下脱去乙酰基,纯化得到式(I)所示的化合物。
7.根据权利要求6所述的人参皂苷衍生物的制备方法,其特征在于,所述式(II)所示的化合物、式(V-1)所示的糖给体或式(V-2)所示的糖给体与路易斯酸的摩尔比为1:1.2~5.0:0.05~0.5;
和/或所述式(III)所示的化合物、式(VI-1)所示的葡萄糖衍生物或式(VI-2)所示的葡萄糖衍生物与路易斯酸的摩尔比为1:2.0~5.0:0.05~0.5;
和/或所述有机溶剂选自吡啶、氯仿、二氯甲烷、乙酸乙酯、四氢呋喃、1,4-二氧六环、氯代苯、甲苯、乙腈、N,N-二甲基甲酰胺或二甲基亚砜;
和/或所述脱硅基保护基试剂选自四甲基氟化铵、四乙基氟化铵、四丁基氟化铵(TBAF)或醋酸;
和/或所述脱烯丙基金属催化剂选自氯化钯(PdCl2)、醋酸钯(Pd(OAc)2)或四三苯基膦钯(Pd(PPh3)4);
和/或所述路易斯酸选自分子筛、三氟化硼乙醚(BF3·Et2O)、三氟甲磺酸三甲基硅脂(TMSOTf)、氯化金(AuCl)、三氯化金(AuCl3)、三苯基膦金(I)三氟甲烷磺酸盐(Ph3PAuOTf)或三苯基膦双(三氟甲磺酰亚胺)金(I)(Ph3PAuNTf2),所述分子筛选自分子筛或酸洗分子筛,底物与分子筛的质量比为1:2.0~5.0;
和/或所述碱为甲醇钠(MeONa)、氢氧化钠(NaOH)或氢氧化钾(KaOH);
和/或所述式(II)所示的化合物与式(V-1)所示的糖给体或式(V-2)所示的糖给体的反应在低温或室温条件下进行,所述低温的温度在-78℃~0℃之间。
8.根据权利要求5所述的人参皂苷衍生物的制备方法,其特征在于,其特征在于,所述式(IV)所示化合物为:
9.一种如权利要求1至权利要求4任一项所述的人参皂苷衍生物的应用,其特征在于,所述人参皂苷衍生物在制备疫苗中的应用。
10.根据权利要求9所述的人参皂苷衍生物的应用,其特征在于,式(I)所述的化合物及其立体异构体或其药学上可接受的盐在制备疫苗中作为佐剂的应用。
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