CN109608513B - 18β-甘草次酸氨基甲酸酯衍生物及其制备方法和应用 - Google Patents

18β-甘草次酸氨基甲酸酯衍生物及其制备方法和应用 Download PDF

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CN109608513B
CN109608513B CN201811581719.3A CN201811581719A CN109608513B CN 109608513 B CN109608513 B CN 109608513B CN 201811581719 A CN201811581719 A CN 201811581719A CN 109608513 B CN109608513 B CN 109608513B
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蔡东
孙玉琦
杨殿深
宫益霞
贾云宏
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Abstract

本发明公开了一种18β‑甘草次酸氨基甲酸酯衍生物，具有如下式(Ⅰ)或式(Ⅱ)所示结构：

Description

18β-甘草次酸氨基甲酸酯衍生物及其制备方法和应用
技术领域
本发明属于药物化学领域，尤其是涉及一种18β-甘草次酸氨基甲酸酯衍生物及其制备方法和应用。
背景技术
三萜酸类化合物多数为四环三萜酸和五环三萜酸，其中五环三萜酸类成分在药用植物中较为常见结构上可分为四类，分别为乌苏烷型、齐墩果烷型、羽扇豆烷型和木栓烷型。甘草次酸(glycyrrhetinic acid，GA)属于五环三萜类化合物，骨架11位上有C＝O，类似18-H-齐墩果烷型结构。
甘草次酸结构存在不同的光学异构体如18α型、18β型，这些异构体的药效作用并不完全相同，其中18α-甘草次酸在药物的肝靶向性、抗炎等方面较18β-甘草次酸更为出色[Beaton J M,Spring F S.J.Chem.Soc.,1955:3126-3129.]，但是由于自然界中18α异构体的量极少，仅占甘草次酸天然总量的3％左右，故甘草次酸及其衍生物的开发研究仍以18β异构体为主。18β-甘草次酸也具有广泛的药理活性，包括类固醇样作用、抗炎抗过敏作用、保肝解毒活性、抗肿瘤作用、抗动脉粥样硬化作用等。
Elsayed等[Elsayed H E,Akl M R,Ebrahim H Y,et al.Chemical Biology&DrugDesign,2015,85(2):231-243.]报道了一系列齐墩果酸的C-3位氨基甲酸酯衍生物(如图-1所示)具有抑制人乳腺癌细胞MDA-MB-231迁移和入侵的作用。Western blot实验分析此类化合物可能靶向Brk/Paxillin/Rac1信号通路。
Ashour等[Ashour A,El-Sharkawy S,Amer M,et al.Bioorganic&MedicinalChemistry,2014,22(1):211-220.]也报道了一系列齐墩果酸的C-3位的氨基甲酸酯衍生物(如式(a)所示)具有抗肿瘤活性。此类化合物可能抑制拓扑异构酶I和II。
Figure GDA0002388955380000011
Bar等[Bar F M,Khanfar M A,Elnagar A Y,et al.Journal of NaturalProducts,2009,72(9):1643-1650.]报道了桦木酸的C-3位的氨基甲酸酯衍生物(如式(b)所示)对人结肠癌细胞SW948和HCT-116以及乳腺癌细胞系MDA-MB-231表现出更好的细胞毒活性。此类化合物很好的抑制拓扑异构酶I和II。
Kommera等[Kommera H,
Figure GDA0002388955380000023
G N,Dittrich S,et al.Bioorganic&Medicinal Chemistry Letters,2010,20(11):3409-3412.]也报道了一系列桦木酸的C-3位的氨基甲酸酯衍生物(如式(b)所示)，对十五种肿瘤细胞株(8505C、SW1736(anaplasticthyroid)、A253、FaDu(head and neck)、A431(cervical)、A2780(ovarian)、DLD-1、HCT-8、HCT-116、HT-29、SW480(colon)、MCF-7(breast)、518A2(melanoma)、A549、(lung)和liposarcoma(connective tissue))进行了筛选检测，部分化合物具有抗增殖活性。
Figure GDA0002388955380000021
Jana等[Jana W,Lucie H,Vincent P,et al.European Journal of MedicinalChemistry,2015,106:194-210.]也报道了一系列桦木酸和桦木醇C-3位的氨基甲酸酯衍生物(如式(c)所示)，对几种人癌细胞(518A2(melanoma)、A2780(ovarian carcinoma)、A549(alveolar basal epithelial adenocarcinoma)、MCF7(breast adenocarcinoma)和线和非恶性小鼠成纤维细胞(NIH 3T3)具有一定的抑制作用。
Figure GDA0002388955380000022
胡立宏等[胡立宏,果德安,雷敏,等.CN 103570792.2014.]报道了一系列蟾毒灵衍生物的C-3位的氨基甲酸酯衍生物(如式(d)所示)，对恶性肿瘤(肝癌、肺癌、乳腺癌、胃癌、食道癌、结肠癌、白血病、淋巴癌、前列腺癌、肾癌等)的细胞株具有抑制活性。
Figure GDA0002388955380000031
甘草次酸对肿瘤具有凋亡作用，利用磺酰罗丹明B(Sulforhodamine B，SRB)比色法检测甘草次酸对细胞增殖的抑制作用，发现甘草次酸对黑色素瘤(518A2)，宫颈癌(A431)，肺癌(A549)，卵巢癌(A2780)，结肠癌(DLD-1，HCT-8，HCT-116，HT-29)，甲状腺癌(8505C，SW-1736)，乳腺癌(MCF-7)和脂肪肉瘤细胞的IC50在74.57-86.50μM之间(即35.10-40.71μg/mL)[RenéCsuk,Schwarz S,Kluge R,et al.Archiv der Pharmazie,2012,345(1):28-32.]。甘草次酸衍生物可通过多种机制对人类肝细胞癌、胃癌、乳腺癌、肺癌、直肠癌、黑色素瘤等多种肿瘤细胞产生不同程度的拮抗作用[Lallemand B,Gelbcke M,DuboisJ,et al.Mini Reviews in Medicinal Chemistry,2011,11(10):881-887.Su X,Wu L,HuM,et al.Biomedicine&Pharmacotherapy,2017,95:670-678.Xu B,Wu G R,Zhang X Y,etal.Molecules,2017,22(6):924-948.]；但是目前所开发的甘草次酸及其衍生物多数药理活性尚不能令人完全满意。因此，有必要进一步开发甘草次酸衍生物来满足患者的需求；结合上述式1-4化合物的结构特点，设计合成了一类新的18β-甘草次酸的氨基甲酸酯衍生物，并对该类化合物进行了初步抗肿瘤活性筛选。
发明内容
本发明要解决的第一个技术问题是提供一种18β-甘草次酸氨基甲酸酯衍生物。该衍生物在具有明显的抗肿瘤活性。
本发明要解决的第二个技术问题是提供上述18β-甘草次酸氨基甲酸酯的制备方法。
本发明要解决的第三个技术问题是提供上述18β-甘草次酸氨基甲酸酯的应用。
为解决上述第一个技术问题，发明一种18β-甘草次酸氨基甲酸酯，具有如下式(Ⅰ)或式(Ⅱ)所示结构：
Figure GDA0002388955380000041
式中：
R选自
Figure GDA0002388955380000042
Figure GDA0002388955380000043
Figure GDA0002388955380000044
中的一种。
为解决上述第二个技术问题，本发明上述式(Ⅰ)所示18β-甘草次酸氨基甲酸酯的制备方法，包括如下步骤：
S10、将18β-甘草次酸(1)和取代的异氰酸酯加入有机溶剂中，回流反应，用薄层色谱监控反应进程；
S11、待反应结束后，将反应混合物减压浓缩，并将得到的残余物通过柱层析法纯化，即得产物(Ⅰ)所示化合物；化学反应结构式如下：
Figure GDA0002388955380000045
优选地，步骤S10中，所述有机溶剂是乙酸乙酯、二氯甲烷、氯仿(三氯甲烷)、乙腈、甲苯、二甲苯中的一种或几种。
优选地，步骤S10中，反应温度为10-120℃；反应时间为12-36h。
本发明上述式(Ⅱ)所示18β-甘草次酸氨基甲酸酯的制备方法，包括如下步骤：
S20、在EDCl、HOBt或Et3N的催化下，将式(1)所示18β-甘草次酸和吗啉在有机溶剂中加热反应，反应结束后，减压蒸馏得浆状物，倒入水中搅拌，抽滤，水洗得化合物(3)；
S21、再将化合物(2)和取代的异氰酸酯加入有机溶液中，回流反应，用薄层色谱监控反应进程。待反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，即得产物(Ⅱ)所示化合物；化学反应方程式如下：
Figure GDA0002388955380000051
优选地，步骤S20中，所述有机溶剂是乙酸乙酯、二氯甲烷、氯仿(三氯甲烷)、乙腈、N,N-二甲基甲酰胺中的一种或几种。
优选地，步骤S21中，所述有机溶剂是乙酸乙酯、二氯甲烷、氯仿(三氯甲烷)、乙腈、甲苯、二甲苯中的一种或几种。
优选地，步骤S21中，所述取代的异氰酸酯中的取代基团选自
Figure GDA0002388955380000052
Figure GDA0002388955380000053
Figure GDA0002388955380000054
中的一种。
为解决上述第三个技术问题，本发明上述式(Ⅰ)或式(Ⅱ)所示18β-甘草次酸氨基甲酸酯化合物及药学上可接受的盐在抗肿瘤药物中的应用。
优选地，所述式(Ⅰ)或式(Ⅱ)所示18β-甘草次酸氨基甲酸酯化合物及药学上可接受的盐在治疗肺癌、肝癌、结肠癌、乳腺癌、前列腺癌的药物中的应用。
本发明所记载的任何范围包括端值以及端值之间的任何数值以及端值或者端值之间的任意数值所构成的任意子范围。
如无特殊说明，本发明中的各原料均可通过市售购买获得，本发明中所用的设备可采用所属领域中的常规设备或参照所属领域的现有技术进行。
与现有技术相比较，本发明具有如下有益效果：
本发明所述化合物结构全新，部分化合物抗肿瘤活性接近克唑替尼，且不会引起高血压、低血钾，盐皮质激素样作用等副作用
具体实施方式
为了更清楚地说明本发明，下面结合优选实施例对本发明做进一步的说明。本领域技术人员应当理解，下面所具体描述的内容是说明性的而非限制性的，不应以此限制本发明的保护范围。
实施例1:
如下结构式所示3β-(((3,4-二氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰa)的制备方法，包括如下步骤：
Figure GDA0002388955380000061
将18β-甘草次酸(0.30mmol)和3,4-二氯苯基异氰酸酯(0.36mmol)加入乙酸乙酯中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体，收率78.1％；1H NMR(400MHz,Chloroform-d)δ7.63(s,1H,phenyl-H),7.33(d,J＝8.7Hz,1H,phenyl-H),7.18(d,J＝8.8Hz,1H,phenyl-H),6.71(s,1H,N-H),5.70(s,1H,CH-12),4.50(dd,J＝10.5,5.9Hz,1H,CH-3),2.87–2.77(m,1H,CH-1),2.37(s,1H,CH-9),2.18(dd,J＝13.8,4.1Hz,1H,CH-16),2.06–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.93(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(d,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C37H50Cl2NO5:658.30660,found:658.31669。
实施例2:
如下结构式所示3β-(((4-氯-3-(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰb)的制备方法，包括如下步骤：
Figure GDA0002388955380000062
将18β-甘草次酸(0.30mmol)和4-氯-3-(三氟甲基)苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化；得白色粉末状固体，收率87.0％；1H NMR(400MHz,Chloroform-d)δ7.76(s,1H,phenyl-H),7.54(s,1H,phenyl-H),7.40(d,J＝8.7Hz,1H,phenyl-H),6.83(s,1H,N-H),5.70(s,1H,CH-12),4.56–4.47(m,1H,CH-3),2.82(d,J＝13.6Hz,1H,CH-1),2.37(s,1H,CH-9),2.17(d,J＝11.9Hz,1H,CH-16),2.06–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(d,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C38H50ClF3NO5:692.33296,found:692.33792。
实施例3:
如下结构式所示3β-(((3,5-二氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰc)的制备方法，包括如下步骤：
Figure GDA0002388955380000071
将18β-甘草次酸(0.30mmol)和3,5-二氯苯基异氰酸酯(0.36mmol)加入三氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体，收率78.1％；1H NMR(400MHz,Chloroform-d)δ7.35(s,2H,phenyl-H),7.02(t,J＝1.8Hz,1H,phenyl-H),6.62(s,1H,N-H),5.69(s,1H,CH-12),4.50(dd,J＝10.7,5.7Hz,1H,CH-3),2.82(d,J＝13.2Hz,1H,CH-1),2.37(s,1H,CH-9),2.17(d,J＝11.9Hz,1H,CH-16),2.03–0.83(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.79–0.73(m,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C37H50Cl2NO5:658.30660,found:658.31238。
实施例4:
如下结构式所示3β-(((4-氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰd)的制备方法，包括如下步骤：
Figure GDA0002388955380000081
将18β-甘草次酸(0.30mmol)和4-氯苯基异氰酸酯(0.36mmol)加入乙腈中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体，收率77.3％；1H NMR(400MHz,Chloroform-d)δ7.33(d,J＝8.4Hz,2H,phenyl-H),7.25(d,J＝2.6Hz,2H,phenyl-H),6.66(s,1H,N-H),5.70(s,1H,CH-12),4.58–4.43(m,1H,CH-3),2.81(dt,J＝13.5,3.6Hz,1H,CH-1),2.37(s,1H,CH-9),2.23–2.13(m,1H,CH-16),2.06–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(s,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C37H51ClNO5:624.34558,found:624.35065。
实施例5:
如下结构式所示3β-(((3-氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰe)的制备方法，包括如下步骤：
Figure GDA0002388955380000082
将18β-甘草次酸(0.30mmol)和3-氯苯基异氰酸酯(0.36mmol)加入甲苯中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体，收率79.0％；1H NMR(400MHz,Chloroform-d)δ7.51(s,1H,phenyl-H),7.20(d,J＝4.9Hz,2H,phenyl-H),7.00(td,J＝4.5,2.0Hz,1H,phenyl-H),6.69(s,1H,N-H),5.70(s,1H,CH-12),4.50(dd,J＝10.4,6.1Hz,1H,CH-3),2.82(dt,J＝13.6,3.6Hz,1H,CH-1),2.37(s,1H,CH-9),2.18(dd,J＝13.0,3.7Hz,1H,CH-16),2.06-1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.82-0.81(m,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C37H51ClNO5:624.34558,found:624.34960。
实施例6:
如下结构式所示3β-(((3-氯-4-甲基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰf)的制备方法，包括如下步骤：
Figure GDA0002388955380000091
将18β-甘草次酸(0.30mmol)和3-氯-4-甲基苯基异氰酸酯(0.36mmol)加入二甲苯中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率74.3％；1H NMR(400MHz,Chloroform-d)δ7.48(s,1H,phenyl-H),7.11(s,2H,phenyl-H),6.60(s,1H,N-H),5.70(s,1H,CH-12),4.49(t,J＝8.3Hz,1H,CH-3),2.81(dt,J＝13.4,3.6Hz,1H,CH-1),2.37(s,1H,CH-9),2.29(s,3H,phenyl-CH3),2.18(dd,J＝13.3,4.2Hz,1H,CH-16),2.09–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(m,1H,CH-5)；HRMS(m/z):[M+Na]+calcd.For C38H52ClNNaO5:660.34317,found:660.34747。
实施例7:
如下结构式所示3β-(((3,5-二甲基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰg)的制备方法，包括如下步骤：
Figure GDA0002388955380000092
将18β-甘草次酸(0.30mmol)和3,5-二甲基苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率92.1％；1H NMR(400MHz,Chloroform-d)δ7.01(s,2H,phenyl-H),6.68(s,1H,phenyl-H),6.55(s,1H,N-H),5.71(s,1H,CH-12),4.49(t,J＝8.2Hz,1H,CH-3),2.85–2.76(m,1H,CH-1),2.37(s,1H,CH-9),2.27(s,6H,phenyl-CH3),2.18(dd,J＝13.5,4.1Hz,1H,CH-16),2.05–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(s,1H,CH-5)；HRMS(m/z):[M+Na]+calcd.For C39H55NNaO5:640.39779,found:640.340185。
实施例8:
如下结构式所示3β-(((4-溴苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰh)的制备方法，包括如下步骤：
Figure GDA0002388955380000101
将18β-甘草次酸(0.30mmol)和4-溴苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率87.2％；1H NMR(400MHz,Chloroform-d)δ7.42–7.35(m,2H,phenyl-H),7.27(d,J＝8.4Hz,2H,phenyl-H),6.65(s,1H,N-H),5.70(s,1H,CH-12),4.50(t,J＝8.3Hz,1H,CH-3),2.85–2.77(m,1H,CH-1),2.37(s,1H,CH-9),2.18(dd,J＝13.7,4.1Hz,1H,CH-16),2.06–1.00(m,17H),1.37(s,3H,CH3-27),1.21(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(s,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C37H51BrNO5:668.29506,found:668.30334,670.30211。
实施例9:
如下结构式所示3β-(((4-氟苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰi)的制备方法，包括如下步骤：
Figure GDA0002388955380000102
将18β-甘草次酸(0.30mmol)和4-氟苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率88.6％；1H NMR(400MHz,DMSO-d6)δ12.25(s,1H,COOH),9.57(s,1H,N-H),7.51(s,2H,phenyl-H),7.14(t,J＝8.9Hz,2H,phenyl-H),5.44(s,1H,CH-12),4.42(dd,J＝11.8,4.6Hz,1H,CH-3),2.73–2.65(m,1H,CH-1),2.46(s,1H,CH-9),2.18–2.04(m,2H,CH-16,CH-2),1.84–0.79(m,17H),1.41(s,3H,CH3-27),1.13(s,3H,CH3-25),1.11(s,3H,CH3-26),1.08(s,3H,CH3-29),0.92(s,3H,CH3-23),0.91(s,3H,CH3-24),0.79(s,3H,CH3-28)；HRMS(m/z):[M+H]+calcd.For C37H51FNO5:608.37513,found:608.38190。
实施例10:
如下结构式所示3β-(((4-(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰj)的制备方法，包括如下步骤：
Figure GDA0002388955380000111
将18β-甘草次酸(0.30mmol)和4-(三氟甲基)苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应。反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化。得白色粉末状固体,收率90.8％；1H NMR(400MHz,Chloroform-d)δ7.52(q,J＝8.7Hz,4H,phenyl-H),6.87(s,1H,N-H),5.71(s,1H,CH-12),4.52(dd,J＝10.5,5.8Hz,1H,CH-3),2.82(dt,J＝13.6,3.6Hz,1H,CH-1),2.37(s,1H,CH-9),2.18(dd,J＝13.5,4.1Hz,1H,CH-16),2.06–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.95(s,3H,CH3-23),0.89(s,3H,CH3-24),0.82(s,3H,CH3-28),0.81(s,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C38H51F3NO5:658.37193,found:658.37843.
实施例11:
如下结构式所示3β-(((3-(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰk)的制备方法，包括如下步骤：
Figure GDA0002388955380000112
将18β-甘草次酸(0.30mmol)和3-(三氟甲基)苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率89.2％；1H NMR(400MHz,Chloroform-d)δ7.72(s,1H,phenyl-H),7.40(t,J＝8.0Hz,1H,phenyl-H),7.28(d,J＝7.8Hz,1H,phenyl-H),6.82(s,1H,N-H),5.71(s,1H,CH-12),4.52(t,J＝8.3Hz,1H,CH-3),2.86–2.78(m,1H,CH-1),2.37(s,1H,CH-9),2.23–2.13(m,1H,CH-16),2.08–1.00(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.95(s,3H,CH3-23),0.89(s,3H,CH3-24),0.82(s,3H,CH3-28),0.81(s,1H,CH-5)；HRMS(m/z):[M+H]+calcd.ForC38H51F3NO5:658.37193,found:658.37843。
实施例12:
如下结构式所示3β-(((3,5-双(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰl)的制备方法，包括如下步骤：
Figure GDA0002388955380000121
将18β-甘草次酸(0.30mmol)和3,5-双(三氟甲基)苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率87.0％；1H NMR(400MHz,Chloroform-d)δ7.90(s,2H,phenyl-H),7.53(s,1H,phenyl-H),6.97(s,1H,N-H),5.70(s,1H,CH-12),4.53(dd,J＝10.6,5.9Hz,1H,CH-3),2.84(dd,J＝10.2,3.4Hz,1H,CH-1),2.37(s,1H,CH-9),2.22–2.13(m,1H,CH-16),2.06–1.01(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.13(s,3H,CH3-29),0.95(s,3H,CH3-23),0.89(s,3H,CH3-24),0.82(s,3H,CH3-28),0.81(s,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C39H50F6NO5:726.35932,found:726.36406。
实施例13:
如下结构式所示3β-(((3-甲氧基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰm)的制备方法，包括如下步骤：
Figure GDA0002388955380000122
将18β-甘草次酸(0.30mmol)和3-甲氧基苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率86.9％；1H NMR(400MHz,Chloroform-d)δ7.17(t,J＝8.2Hz,1H,phenyl-H),6.84(d,J＝8.0Hz,1H,phenyl-H),6.65(s,1H,N-H),6.59(dd,J＝8.4,2.4Hz,1H,phenyl-H),5.70(s,1H,CH-12),4.55–4.45(m,1H,CH-3),3.79(s,3H,CH3),2.85–2.76(m,1H,CH-1),2.37(s,1H,CH-9),2.18(dd,J＝13.4,4.1Hz,1H,CH-16),2.05–1.00(m,17H),1.37(s,3H,CH3-27),1.21(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(s,1H,CH-5)；HRMS(m/z):[M+Na]+calcd.For C38H53NNaO6:642.37706,found:642.37890。
实施例14:
如下结构式所示3β-(((4-甲氧基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰn)的制备方法，包括如下步骤：
Figure GDA0002388955380000131
将18β-甘草次酸(0.30mmol)和4-甲氧基苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率89.3％；1H NMR(400MHz,Chloroform-d)δ7.28(s,2H,phenyl-H),6.83(d,J＝8.9Hz,2H,phenyl-H),6.50(s,1H,N-H),5.70(s,1H,CH-12),4.48(t,J＝8.2Hz,1H,CH-3),3.76(s,3H,CH3),2.80(dt,J＝13.5,3.7Hz,1H,CH-1),2.37(s,1H,CH-9),2.22–2.13(m,1H,CH-16),2.06–0.99(m,17H),1.36(s,3H,CH3-27),1.21(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.93(s,3H,CH3-23),0.86(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(s,1H,CH-5)；HRMS(m/z):[M+Na]+calcd.For C38H53NNaO6:642.37706,found:642.38301。
实施例15:
如下结构式所示3β-(((4-(三氟甲氧基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-酸(简称：Ⅰo)的制备方法，包括如下步骤：
Figure GDA0002388955380000132
将18β-甘草次酸(0.30mmol)和4-(三氟甲氧基)苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化，得白色粉末状固体,收率92.1％；1H NMR(400MHz,Chloroform-d)δ7.40(d,J＝8.5Hz,2H,phenyl-H),7.14(d,J＝8.6Hz,2H,phenyl-H),6.72(s,1H,N-H),5.70(s,1H,CH-12),4.58–4.42(m,1H,CH-3),2.85–2.76(m,1H,CH-1),2.37(s,1H,CH-9),2.18(dd,J＝13.5,4.1Hz,1H,CH-16),2.05–1.00(m,17H),1.36(s,3H,CH3-27),1.21(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.82(s,3H,CH3-28),0.80(s,1H,CH-5)；HRMS(m/z):[M+H]+calcd.For C38H51F3NO6:674.36685,found:674.37311。
实施例16:
如下结构式所示3β-(((3,4-二氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱa)的制备方法，包括如下步骤：
Figure GDA0002388955380000141
18β-甘草次酸(0.47g，1.0mmol)和EDCl(0.23g,1.2mmol),Et3N(0.13g,1.2mmol),HOBt(0.16g,1.2mmol)依次加到乙腈中加热反应20min，然后吗啉(0.11g,1.2mmol)加到上述溶液中，继续回流反应，用薄层色谱监控反应进程；待反应结束后，减压蒸馏得浆状物，倒入水中搅拌，抽滤，水洗得化合物(2)；再将化合物(2)(0.30mmol)和3,4-二氯苯基异氰酸酯(0.36mmol)加入二氯甲烷中回流反应；反应结束后，将混合物减压浓缩，并将得到的残余物通过柱层析法纯化；得白色粉末状固体,收率93.0％；1H NMR(400MHz,Chloroform-d)δ7.63(s,1H,phenyl-H),7.32(d,J＝8.8Hz,1H,phenyl-H),7.18(d,J＝8.8Hz,1H,phenyl-H),6.72(s,1H,N-H),5.68(s,1H,CH-12),4.48(dd,J＝10.8,5.7Hz,1H,CH-3),3.65(t,J＝6.3Hz,8H,morpholine-H),2.81(dt,J＝13.4,3.8Hz,1H,CH-1),2.35(s,1H,CH-9),2.27(d,J＝12.7Hz,1H,CH-16),2.05–0.99(m,17H),1.35(s,3H,CH3-27),1.20(s,3H,CH3-25),1.14(s,3H,CH3-26),1.10(s,3H,CH3-29),0.93(s,3H,CH3-23),0.86(s,3H,CH3-24),0.80(s,3H,CH3-28),0.79(s,1H,CH-5)；HRMS(m/z):[M+Na]+ calcd. For C41H56Cl2N2NaO5: 749.34640,found: 749.34901.
实施例17:
如下结构式所示3β-(((3,5-二氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱc)的制备方法，包括如下步骤：
Figure GDA0002388955380000151
以与实施例16相同的方法制备，其不同之处仅在于：3,5-二氯苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率91.7％；1H NMR(400MHz,Chloroform-d)δ7.35(s,2H,phenyl-H),7.01(q,J＝2.7,1.9Hz,1H,phenyl-H),6.78(s,1H,N-H),5.68(s,1H,CH-12),4.48(dd,J＝10.9,5.5Hz,1H,CH-3),3.65(q,J＝6.4,5.8Hz,8H,morpholine-H),2.81(dt,J＝13.4,3.4Hz,1H,CH-1),2.35(s,1H,CH-9),2.31–2.22(m,1H,CH-16),2.07–1.00(m,17H),1.35(s,3H,CH3-27),1.21(s,3H,CH3-25),1.14(s,3H,CH3-26),1.11(s,3H,CH3-29),0.93(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C41H56Cl2N2NaO5:749.34640,found:749.34951。
实施例18:
如下结构式所示3β-(((4-氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱd)的制备方法，包括如下步骤：
Figure GDA0002388955380000152
以与实施例16相同的方法制备，其不同之处仅在于：4-氯苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率92.8％；1H NMR(400MHz,Chloroform-d)δ7.33(d,J＝8.6Hz,2H,phenyl-H),7.24(d,J＝7.8Hz,2H,phenyl-H),6.64(s,1H,N-H),5.68(s,1H,CH-12),4.48(dd,J＝10.6,5.9Hz,1H,CH-3),3.67-3.56(m,8H,morpholine-H),2.80(dt,J＝13.6,3.6Hz,1H,CH-1),2.35(s,1H,CH-9),2.31–2.22(m,1H,CH-16),2.07–0.99(m,17H),1.35(s,3H,CH3-27),1.20(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.93(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C41H57ClN2NaO5:715.38537,found:715.38855。
实施例19:
如下结构式所示3β-(((3-氯苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱe)的制备方法，包括如下步骤：
Figure GDA0002388955380000161
以与实施例16相同的方法制备，其不同之处仅在于：3-氯苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率94.4％；1H NMR(400MHz,Chloroform-d)δ7.52(s,1H,phenyl-H),7.19(d,J＝5.1Hz,2H,phenyl-H),7.00(dq,J＝7.1,2.0Hz,1H,phenyl-H),6.70(s,1H,N-H),5.68(s,1H,CH-12),4.49(dd,J＝10.7,5.9Hz,1H,CH-3),3.69–3.56(m,8H,morpholine-H),2.80(dt,J＝13.6,3.6Hz,1H,CH-1),2.35(s,1H,CH-9),2.27(d,J＝13.2Hz,1H,CH-16),2.09–0.99(m,17H),1.35(s,3H,CH3-27),1.20(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.93(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C41H57ClN2NaO5:715.38537,found:715.38920。
实施例20:
如下结构式所示3β-(((3-氯-4-甲基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱf)的制备方法，包括如下步骤：
Figure GDA0002388955380000162
以与实施例16相同的方法制备，其不同之处仅在于：3-氯-4-甲基苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率91.8％；1H NMR(400MHz,Chloroform-d)δ7.50(s,1H,phenyl-H),7.12(s,2H,phenyl-H),6.57(s,1H,N-H),5.69(s,1H,CH-12),4.49(dd,J＝10.3,6.1Hz,1H,CH-3),3.64(qd,J＝8.5,8.1,3.5Hz,8H,morpholine-H),2.81(dt,J＝13.6,3.6Hz,1H,CH-1),2.36(s,1H,CH-9),2.33–2.28(m,3H,CH3),2.25(d,J＝3.2Hz,1H,CH-16),2.08–1.00(m,17H),1.36(s,3H,CH3-27),1.21(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.83(s,1H,CH-5),0.81(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C42H59ClN2NaO5:729.40102,found:729.40510。
实施例21:
如下结构式所示3β-(((4-溴苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱg)的制备方法，包括如下步骤：
Figure GDA0002388955380000171
以与实施例16相同的方法制备，其不同之处仅在于：4-溴苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率90.6％；1H NMR(400MHz,Chloroform-d)δ7.38(dd,J＝8.9,2.1Hz,2H,phenyl-H),7.28(d,J＝8.4Hz,2H,phenyl-H),6.66(s,1H,N-H),5.68(d,J＝1.9Hz,1H,CH-12),4.48(dd,J＝11.3,5.5Hz,1H,CH-3),3.73–3.55(m,8H,morpholine-H),2.85–2.75(m,1H,CH-1),2.35(s,1H,CH-9),2.27(d,J＝12.8Hz,1H,CH-16),2.07–0.99(m,17H),1.35(s,3H,CH3-27),1.20(s,3H,CH3-25),1.14(s,3H,CH3-26),1.10(s,3H,CH3-29),0.93(s,3H,CH3-23),0.87(s,3H,CH3-24),0.82(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C41H57BrN2NaO5:759.33486,found:759.33486,761.33783。
实施例22:
如下结构式所示3β-(((4-氟苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱh)的制备方法，包括如下步骤：
Figure GDA0002388955380000172
以与实施例16相同的方法制备，其不同之处仅在于：4-氟苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率94.0％；1H NMR(400MHz,Chloroform-d)δ7.33(s,2H,phenyl-H),6.97(td,J＝8.6,1.5Hz,2H,phenyl-H),6.62(s,1H,N-H),5.67(s,1H,CH-12),4.48(dd,J＝10.3,6.1Hz,1H,CH-3),3.65-3.59(m,8H,morpholine-H),2.80(dt,J＝14.1,3.6Hz,1H,CH-1),2.35(s,1H,CH-9),2.31–2.22(m,1H,CH-16),2.05–0.99(m,17H),1.35(s,3H,CH3-27),1.20(s,3H,CH3-25),1.14(s,3H,CH3-26),1.11(s,3H,CH3-29),0.93(s,3H,CH3-23),0.86(s,3H,CH3-24),0.82(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C41H58FN2O5:677.43298,found:677.43850。
实施例23:
如下结构式所示3β-(((4-(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱi)的制备方法，包括如下步骤：
Figure GDA0002388955380000181
以与实施例16相同的方法制备，其不同之处仅在于：4-(三氟甲基)苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率91.4％；1H NMR(400MHz,Chloroform-d)δ7.52(q,J＝8.8Hz,4H,phenyl-H),6.83(s,1H,N-H),5.68(s,1H,CH-12),4.51(dd,J＝10.8,5.6Hz,1H,CH-3),3.70–3.56(m,8H,morpholine-H),2.82(dt,J＝13.6,3.6Hz,1H,CH-1),2.35(s,1H,CH-9),2.28(dd,J＝13.9,3.6Hz,1H,CH-16),2.08–0.99(m,17H),1.35(s,3H,CH3-27),1.21(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.83(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C42H57F3N2NaO5:749.41173,found:749.41688。
实施例24:
如下结构式所示3β-(((3-(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱj)的制备方法，包括如下步骤：
Figure GDA0002388955380000182
以与实施例16相同的方法制备，其不同之处仅在于：3-(三氟甲基)苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率91.5％；1H NMR(400MHz,Chloroform-d)δ7.76(s,1H,phenyl-H),7.56(d,J＝8.1Hz,1H,phenyl-H),7.41(t,J＝8.0Hz,1H,phenyl-H),7.30(d,J＝7.7Hz,1H,phenyl-H),6.86(s,1H,N-H H),5.70(s,1H,CH-12),4.53(dd,J＝10.5,6.0Hz,1H,CH-3),3.66(dd,J＝10.9,5.4Hz,8H,morpholine-H),2.83(dt,J＝13.7,3.7Hz,1H,CH-1),2.37(s,1H,CH3-27),2.34–2.25(m,1H,CH-9),2.10–1.02(m,17),1.37(s,3H),1.23(s,3H,CH3-25),1.17(s,3H,CH3-26),1.13(s,3H,CH3-29),0.96(s,3H,CH3-23),0.90(s,3H,CH3-24),0.85(s,1H,CH-5),0.82(s,3H,CH3-28)；HRMS(m/z):[M+H]+calcd.For C42H58F3N2O5:727.42978,found:727.43507。
实施例25:
如下结构式所示3β-(((3,5-双(三氟甲基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱk)的制备方法，包括如下步骤：
Figure GDA0002388955380000191
以与实施例16相同的方法制备，其不同之处仅在于：3,5-双(三氟甲基)苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率88.9％；1H NMR(400MHz,Chloroform-d)δ7.92(s,2H,phenyl-H),7.52(s,1H,phenyl-H),7.16(d,J＝2.5Hz,1H,N-H),5.69(d,J＝2.5Hz,1H,CH-12),4.52(dd,J＝11.3,5.7Hz,1H,CH-3),3.68–3.64(m,8H,morpholine-H),2.82(dd,J＝13.7,3.3Hz,1H,CH-1),2.35(s,1H,CH-9),2.28(d,J＝13.6Hz,1H,CH-16),2.08–1.00(m,17H),1.35(s,3H,CH3-27),1.21(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.83(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C43H56F6N2NaO5:817.39911,found:817.40464。
实施例26:
如下结构式所示3β-(((3-甲氧基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱl)的制备方法，包括如下步骤：
Figure GDA0002388955380000192
以与实施例16相同的方法制备，其不同之处仅在于：3-甲氧基苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率92.2％；1H NMR(400MHz,Chloroform-d)δ7.14(s,1H,phenyl-H),6.85(d,J＝8.1Hz,1H,phenyl-H),6.62(s,1H,N-H),6.60–6.57(m,1H,phenyl-H),5.67(s,1H,CH-12),4.49(dd,J＝10.4,6.1Hz,1H,CH-3),3.81–3.75(m,3H,CH3),3.68–3.55(m,8H,morpholine-H),2.85–2.75(m,1H,CH-1),2.35(s,1H,CH-9),2.31–2.22(m,1H,CH-16),2.07–0.99(m,17H),1.35(s,3H,CH3-27),1.21(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.94(s,3H,CH3-23),0.88(s,3H,CH3-24),0.83(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C42H60N2NaO6:711.43491,found:711.44018.
实施例27:
如下结构式所示3β-(((4-甲氧基苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱm)的制备方法，包括如下步骤：
Figure GDA0002388955380000201
以与实施例16相同的方法制备，其不同之处仅在于：4-甲氧基苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率92.6％；1H NMR(400MHz,Chloroform-d)δ7.28(s,2H,phenyl-H),6.86–6.79(m,2H,phenyl-H),6.50(s,1H,N-H),5.67(d,J＝1.7Hz,1H,CH-12),4.47(t,J＝8.4Hz,1H,CH-3),3.76(d,J＝1.4Hz,3H,CH3),3.70–3.55(m,8H,morpholine-H),2.79(d,J＝13.5Hz,1H,CH-1),2.35(s,1H,CH-9),2.30–2.21(m,1H,CH-16),2.15–0.99(m,17H),1.35(s,3H,CH3-27),1.21(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-29),0.93(s,3H,CH3-23),0.86(s,3H,CH3-24),0.82(s,1H,CH-5),0.80(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C42H60N2NaO6:711.43491,found:711.43961。
实施例28:
如下结构式所示3β-(((4-(三氟甲氧基)苯基)氨基甲酰基)氧基)-11-氧代-齐墩果-12-烯-29-甲酰基吗啉(简称：Ⅱn)的制备方法，包括如下步骤：
Figure GDA0002388955380000211
以与实施例16相同的方法制备，其不同之处仅在于：4-(三氟甲氧基)苯基异氰酸酯替代3,4-二氯苯基异氰酸酯；得白色粉末状固体,收率94.0％；1H NMR(400MHz,Chloroform-d)δ7.43(d,J＝8.5Hz,2H,phenyl-H),7.16(d,J＝8.4Hz,2H,phenyl-H),6.75(s,1H,N-H),5.70(d,J＝1.9Hz,1H,CH-12),4.51(dd,J＝11.2,5.5Hz,1H,CH-3),3.67(d,J＝5.3Hz,8H，morpholine-H),2.87–2.78(m,1,CH-1H),2.37(s,1H,CH-9),2.29(d,J＝12.8Hz,1H,CH-16)),2.05–0.99(m,17H),1.37(s,3H,CH3-27),1.22(s,3H,CH3-25),1.17(s,3H,CH3-26),1.13(s,3H,CH3-29),0.96(s,3H,CH3-23),0.89(s,3H,CH3-24),0.85(s,1H,CH-5),0.82(s,3H,CH3-28)；HRMS(m/z):[M+Na]+calcd.For C42H57F3N2NaO6:765.40664,found:765.41272。
实验效果检测实验:18β-甘草次酸的氨基甲酸酯衍生物的抗肿瘤活性测定
本发明化合物作为抗肿瘤抑制剂的活性可以按许多标准的生物学试验或药理学试验测定。
肿瘤细胞培养于RPMI1640完全培养液中，置37℃、5％CO2培养箱中培养，取对数生长期细胞用胰酶/EDTA消化液消化，离心，用完全培养液吹打成单细胞悬液。调整浓度为5×104个/mL。96孔板每孔准确接种细胞悬液100uL，培养24h，弃去培养液，组各孔分别加入5个不同浓度的受试物100uL，每个浓度3个复孔。培养48h后，弃去旧液，每孔用200uL生理盐水清洗后弃去，MTT储备液用培养液10倍稀释后，各孔加入终浓度为0.5mg/mL的MTT 100uL，继续培养4h。弃去旧液，每孔加入100uL DMSO，培养板震荡3min。用酶标仪测定每孔的吸光度值，A1孔调零，测试波长490，630nm。各组OD值取均值后，计算抑制百分数。用GraphPadPrism 6软件采用非线性回归法计算IC50值。
细胞抑制率＝(1-实验组吸光值/对照组吸光值)×100％
18β-甘草次酸的氨基甲酸酯衍生物的对肿瘤细胞抑制率的测定结果
Figure GDA0002388955380000212
Figure GDA0002388955380000221
Figure GDA0002388955380000231
部分18β-甘草次酸的氨基甲酸酯衍生物的对肿瘤细胞增殖抑制活性
Figure GDA0002388955380000232
备注：8＜a＜40为检测药物浓度只在浓度为40μg/mL时有明显抑制效果，故IC50计算不准确，只能判断为在8-40μg/mL之间，以作参考。
结果表明，本发明涉及的18β-甘草次酸的氨基甲酸酯衍生物对多种人源肿瘤细胞具有明显的增殖抑制活性，都有不同程度的抑制作用。并且通式I所示的化合物抗肿瘤活性明显好于通式Ⅱ所示的化合物。
本发明的化合物的发明点在于：与母核结构18β-甘草次酸比较，本发明设计合成的18β-甘草次酸氨基甲酸酯衍生物结构全新，化合物对肺癌、肝癌、结肠癌、乳腺癌、前列腺癌细胞系具有显著的抑制作用，部分化合物抗肿瘤活性接近克唑替尼。
母核结构18β-甘草次酸(1)的抗肿瘤活性：
A549 90μmol/L(即42.36μg/mL)
HT29 80μmol/L(即37.65μg/mL)
[RenéCsuk,Schwarz S,Kluge R,et al.Archiv der Pharmazie,2012,345(1):28-32.]
A549>100μmol/L(即>47.07μg/mL)
[Lallemand B,Chaix F,Bury M,et al.Journal of medicinal chemistry,2011,54(19):6501-6513.]
显然，本发明的上述实施例仅仅是为清楚地说明本发明所作的举例，而并非是对本发明的实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无法对所有的实施方式予以穷举。凡是属于本发明的技术方案所引伸出的显而易见的变化或变动仍处于本发明的保护范围之列。

Claims (1)

1.如下式(Ⅰ)所示18β-甘草次酸氨基甲酸酯化合物及药学上可接受的盐在抗肝癌或前列腺癌药物中的应用，其特征在于：式(Ⅰ)所示结构如下：
Figure FDA0002526365430000011
式中：
R选自
Figure FDA0002526365430000012
Figure FDA0002526365430000013
Figure FDA0002526365430000014
中的一种。
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