English

• 齐墩果酸(Oleanolic acid, OA)和熊果酸(Ursolic acid, UA)是广泛分布于植物界的三萜类化合物^[1], 其结构如图 1所示. UA具有使癌症发展失调的生物化学活性^[2].同时UA及其衍生物具有多种生物活性, 如抗HIV^[3]、抗肝损伤^[4]、抗疟疾^[5]、抗菌、抗炎^[6]及免疫调节^[7]等活性. UA及其酯化衍生物对一些肿瘤细胞系具有显著的细胞毒活性^[8], UA对癌细胞的起始、增殖和转移起到抑制作用^[9].而UA的同分异构体OA, 也具有相似的药理活性^[10]. OA具有保肝作用, 其通过抑制CYP2E1 (CYP2E1是参与化学药品和药物转化成有毒代谢产物的一种同工酶)的表达而起到保肝的作用^[11]. OA具有抗炎活性, 并对癌细胞的增殖具有抑制作用^[12].但OA和UA的水溶性较差, 进而限制了它们的生物利用度, 故需通过化学修饰来提高其水溶性和活性.陈等^[13]通过在OA和UA的C-28位引入丁二酸片段后, 与环状胺类化合物偶联得到较OA和UA活性更好的衍生物; Tian等^[14]在C-28位引入乙醇酸片段后与链状的N-Boc乙二胺等胺类化合物偶联, 其IC₅₀值明显优于先导化合物和阳性对照物Gefitinib; OA衍生物2-氰基-3, 12-二氧代齐墩果烷-1, 9-二烯-28-羧酸(CDDO)及它的C-28位甲酯(CDDO-Me)、C-28位咪唑(CDDO-Im)均表现出了较OA更强的抗炎和抗肿瘤活性, CDDO-Me已进入临床研究阶段, 其在A环上均具有双键的结构(结构如图 2)^[10].故本工作以此为依据, 在A环的C-2和C-3位间形成双键, 分别通过戊二酸和乙醇酸连接片段, 在C-28位分别导入吗啉、N-甲基哌嗪和N-Boc乙二胺片段, 设计合成了20种未见文献报道的齐墩果酸和熊果酸衍生物, 以期得到高效、低毒和生物活性更高的新型五环三萜类化合物.

  图 1

  

  图 1.  齐墩果酸和熊果酸的结构

  Figure 1.  Structure of oleanolic and ursolic acid

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  图 2

  

  图 2.  CDDO、CDDO-Me和CDDO-Im的结构

  Figure 2.  Structures of CDDO, CDDO-Me and CDDO-Im

  下载: 全尺寸图片 幻灯片

  我们设计在C-28位导入顺丁烯二酸、戊二酸和乙醇酸连接片段, 但接入顺丁烯二酸片段所得产物OA-5经¹H NMR测试, 出现了如下谱图(图 3), 其峰面积比为1:3(图 3左侧峰的偶合常数为15.8 Hz, 右侧的偶合常数为12.8 Hz), 即顺式结构/反式结构物质的量之比为3:1, 结构发生了消旋化.而导入戊二酸和乙醇酸连接片段时(如Scheme 1和Scheme 2)则顺利的得到相应中间体及目标产物.

  图 3

  

  图 3.  OA-5结构及局部¹H NMR图

  Figure 3.  Structure and ¹H NMR partially view of OA-5

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  图式 1

  

  图式 1.  齐墩果酸衍生物OA-6~OA-6d和熊果酸衍生物UA-6~UA-6d的合成

  Scheme 1.  Synthesis of OA derivatives of OA-6~OA-6d and UA derivatives of UA-6~UA-6d

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  图式 2

  

  图式 2.  齐墩果酸衍生物OA-9~OA-9d和熊果酸衍生物UA-9~UA-9d的合成

  Scheme 2.  Synthesis of OA derivatives of OA-6~OA-6d and UA derivatives of UA-9~UA-9d

  下载: 全尺寸图片 幻灯片

  1.   结果与讨论

  1.1   目标化合物的合成

  以齐墩果酸和熊果酸为原料, 设计合成了20种未见文献报道的化合物, 所有的新化合物结构均经IR、¹H NMR、¹³C NMR和HRMS得到确证. IR谱图呈现出正常的特殊官能团吸收峰, ¹H NMR和¹³C NMR谱图呈现正常的偶合裂分, HRMS测得数值与理论值相符.

  1.2   抗肿瘤细胞增殖实验

  对8个中间体及12个目标产物进行了抗人肺腺癌细胞A549、人乳腺癌MCF-7细胞株和人肝肿瘤细胞HepG2细胞株抑制作用测试, 测试结果见表 1.

  表 1

  

  表 1  OA和UA衍生物的抑制A549、MCF-7和HepG2细胞的IC₅₀值

  Table 1.  IC₅₀ of OA and UA derivatives on A549, MCF-7 and HepG2

  下载: 导出CSV

  Sample	MW	IC50/(μmol•L－1)
  		A549	MCF-7	HepG2
  顺铂		2.3±0.19	2.6±0.94	4.7±1.3
  紫杉醇	853.92	96.6±10.4	89.7±6.59	108.6±9.7
  OA	456.70	＞50	＞50	＞50
  OA-6	538.80	9.60±1.87	11.66±1.76	21.24±2.12
  OA-6a	607.91	＞50	＞50	＞50
  OA-6b	619.96	＞50	＞50	42.00±1.58
  OA-6c	681.00	＞50	＞50	＞50
  OA-6d	580.88	39.15±2.95	13.54±1.32	4.87±1.65
  OA-9	496.72	24.81±1.88	34.04±2.01	41.62±4.36
  OA-9a	565.83	＞50	＞50	＞50
  OA-9b	578.87	＞50	36.18±5.52	20.55±1.65
  OA-9c	638.92	＞50	＞50	＞50
  OA-9d	538.80	15.70±3.16	5.25±0.71	4.98±1.22
  UA	456.70	5.81±0.32	9.57±2.55	17.78±1.63
  UA-6	538.80	15.43±2.79	13.79±1.36	23.44±2.14
  UA-6a	607.91	＞50	＞50	＞50
  UA-6b	619.96	＞50	＞50	42.00±3.24
  UA-6c	681.00	＞50	＞50	＞50
  UA-6d	580.88	37.39±12.17	9.16±0.17	8.88±2.50
  UA-9	496.72	14.50±2.01	19.81±1.03	24.74±1.52
  UA-9a	565.83	＞50	＞50	＞50
  UA-9b	578.87	10.05±1.40	12.14±2.05	17.94±1.66
  UA-9c	638.92	＞50	＞50	＞50
  UA-9d	538.80	5.61±3.16	3.52±0.71	4.51±1.22
  a The unit of IC50 value of Taxol is nmol•L－1.

  从表 1可以看出, 合成的20种衍生物中有8种衍生物的抑制活性优于先导化合物OA和UA, 其中, OA-9d和UA-9d对人肺腺癌细胞A549、人乳腺癌MCF-7细胞株和人肝肿瘤细胞HepG2细胞株都显示出很高的抑制活性, 明显优于先导化合物; 而与阳性对照物紫杉醇相比, 其活性弱于阳性对照物; 与阳性对照物顺铂相比, 对于HepG2细胞株UA-9d的活性优于阳性对照物, 而UA-6d和UA-9d与其活性相当.

  1.3   UA-9d体外抗缺氧诱导因子1α (HIF-1α)活性

  选择对三种癌细胞株活性最好的化合物UA-9d进行了抑制肝癌细胞Hep3B的HIF-1α表达研究, 实验检测结果如图 4.

  图 4

  

  图 4.  UA-9d抑制HIF-1α的表达情况

  Figure 4.  UA-9d inhibits the expression of hypoxia-inducible factor 1α

  (－): negative control, under the condition of 20% O₂, dissolve in DMSO; (+): positive control, under the condition of 1% O₂, dissolve in DMSO. **P < 0.01.

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  从图 4可以看出, 随着UA-9d的浓度增大, HIF-1α的表达率逐渐减少, 抑制HIF-1α表达的效果逐渐增强, 当UA-9d的浓度达到10 μmol/L时, 抑制率达到64%左右.结果表明UA-9d能够有效地抑制Hep3B细胞中的HIF-1α的表达.

  2.   结论

  通过活性对比发现以戊二酸为连接片段的关键中间体对三种肿瘤细胞的抑制活性明显优于以乙醇酸作为连接片段的关键中间体; 在C-28位通过连接片段与不同的含氮化合物偶联显示出不同的活性, 连接N-甲基哌嗪衍生物的活性优于连接吗啉的衍生物, 而连接链状的胺类衍生物的活性明显优于连接环状胺类的衍生物, 推测是由于裸露的一级胺使其化合物极性增强, 从而提高其生物活性.以上结果为以后OA和UA的结构修饰改造提供了方向.

  3.   实验部分

  3.1   仪器与试剂

  X-5型显微熔点测定仪(温控型)(北京泰克仪器有限公司); WGH-30型双光束红外分光光度计(上海勇规分析仪器有限公司), KBr压片; BRUKER-500MHz核磁共振仪(瑞士BRUKER公司), CDCl₃为溶剂; GC-Tof MS型高分辨率质谱; Multiskan Ascent型酶标仪(美国Thermo Fisher Scientific).所用药品和试剂均为分析纯或化学纯.

  3.2   齐墩果酸衍生物的合成

  3β-羟基齐墩果-12-烯-28-羧酸甲酯(OA-1)、3β-甲基磺酰氧基齐墩果-12-烯-28-酸甲酯(OA-2)、齐墩果烷- 2, 12-二烯-28-羧酸甲酯(OA-3)、齐墩果烷-2, 12-二烯-28-醇(OA-4)的合成参考文献[15], 均得到白色固体产物. 3β-甲基磺酰氧基齐墩果-12-烯-28-酸(OA-7)、齐墩果烷- 2, 12-二烯-28–酸(OA-8)的合成参考文献[16], 均得到淡黄色固体产物.

  3.2.1   5-氧亚基-5-(齐墩果烷-2, 12-二烯-28-基氧基)戊酸(OA-6)的合成

  将OA-4 (600 mg, 1.41 mmol)、戊二酸酐(323.5 mg, 2.82 mmol)、对二甲氨基吡啶(DMAP, 189.6 mg, 1.69 mmol)和无水CH₂Cl₂依次加入圆底烧瓶中, 加热回流8 h, 反应完毕后用H₂O和CH₂Cl₂对反应混合物进行萃取, 有机层用无水MgSO₄进行干燥, 过滤, 经减压蒸馏的产物, 再经柱层析得到509.5 mg白色固体OA-6, 收率67%. m.p. 64~65 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.43~5.36 (m, 2H, H-2 & H-3), 5.24 (s, 1H, H-12), 4.05 (d, J＝－11.0 Hz, 1H, CHO), 3.75 (d, J＝－11.0 Hz, 1H, CHO), 2.44~2.40 (m, 4H, COCH₂CH₂CH₂CO), 2.06 (dd, J＝3.9, 13.5 Hz, 1H, H-18), 1.17, 0.98, 0.96, 0.90, 0.89, 0.88 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 178.53, 172.91, 143.37, 137.94, 123.18, 121.40, 70.78, 52.03, 46.25, 46.08, 42.63, 41.80, 40.90, 39.88, 36.10, 35.90, 34.45, 34.00, 33.31, 33.10, 32.97, 31.95, 31.78, 31.41, 30.85, 25.81, 25.48, 23.51, 23.46, 22.78, 22.43, 19.96, 19.49, 16.45, 15.70; IR (KBr) ν: 3448, 3012, 2953, 2867, 1740, 1711 cm^－1; HRMS calcd for C₃₅H₅₃O₄ [M－H]^－ 537.3944, found 537.3953.

  3.2.2   5-氧亚基-5-(1-吗啉基)戊酸-(齐墩果烷-2, 12-二烯-28-基)酯(OA-6a)的合成

  将OA-6 (89 mg, 0.17 mmol)、吗啉(0.02 mL, 0.20 mmol)、DMAP (24.4 mg, 0.20 mmol)用CH₂Cl₂溶解, 将反应物冷却至0 ℃, 再将1-乙基-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDCI, 162.9 mg, 0.85 mmol)溶于CH₂Cl₂中(逐滴加入到反应液中)于0 ℃下, 继续搅拌5 min, 然后室温下搅拌3 h.反应完毕后减压蒸馏除去溶剂, 用H₂O和CH₂Cl₂对反应混合物进行萃取, 有机层用无水MgSO₄进行干燥, 过滤, 经减压蒸馏的产物, 再经柱层析得到55.6 mg白色固体OA-6a, 收率55%. m.p. 53~54 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.42~5.35 (m, 2H, H-2 & H-3), 5.22 (s, 1H, H-12), 4.04 (d, J＝－11.0 Hz, 1H, CHO), 3.73 (d, J＝－11.0 Hz, 1H, CHO), 3.65~3.46 (m, 8H, morpholine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.43~2.35 (m, 4H, COCH₂CH₂CH₂CO), 2.05 (dd, J＝3.9, 13.4 Hz, 1H, H-18), 1.16, 0.97, 0.95, 0.89, 0.88, 0.87 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.85, 170.38, 142.91, 137.45, 122.66, 120.89, 70.19, 66.42, 66.16, 51.54, 45.76, 45.58, 45.45, 42.19, 41.33, 40.41, 39.39, 35.61, 35.43, 33.97, 33.52, 33.00, 32.60, 31.56, 31.29, 30.97, 30.37, 25.31, 25.01, 23.05, 22.98, 22.30, 21.91, 19.99, 19.00, 15.99, 15.22; IR (KBr) ν: 3005, 2946, 2925, 2859, 1733, 1646 cm^－1; HRMS calcd for C₃₉H₆₂NO₄ [M+H]⁺ 608.4680, found 608.4667.

  3.2.3   5-氧亚基-5-(4-甲基-1-哌嗪基)戊酸-(齐墩果烷-2, 12-二烯-28-基)酯(OA-6b)的合成

  按照合成OA-6a的方法由化合物将OA-6 (80 mg, 0.15 mmol)、N-甲基哌嗪(0.02 mL, 0.18 mmol)、DMAP (22.0 mg, 0.18 mmol)和EDCI (143.8 mg, 0.75 mmol)反应得到了82.8 mg白色固体产物OA-6b, 收率90%. m.p. 52~53 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.42~5.35 (m, 2H, H-2 & H-3), 5.22 (s, 1H, H-12), 4.04 (d, J＝ －11.0 Hz, 1H, CHO), 3.73 (d, J＝－11.0 Hz, 1H, CHO), 3.62~2.37 (m, 12H, COCH₂CH₂CH₂CO & piperazine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.29 (s, 3H, NCH₃), 2.05 (d, J＝13.0 Hz, 1H, H-18), 1.16, 0.97, 0.89, 0.88, 0.87 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.88, 170.10, 142.92, 137.44, 122.64, 120.89, 70.16, 54.69, 54.23, 51.53, 45.76, 45.58, 45.50, 44.88, 42.19, 41.32, 40.98, 40.41, 39.39, 35.61, 35.42, 33.96, 33.52, 33.10, 32.61, 31.73, 31.48, 31.29, 30.98, 30.37, 25.31, 25.01, 23.06, 22.30, 21.89, 20.09, 19.01, 15.99, 15.22; IR (KBr) ν: 3005, 2946, 2925, 2787, 1733, 1653 cm^－1; HRMS calcd for C₄₀H₆₅N₂O₃ [M+H]⁺ 621.4996, found 621.4980.

  3.2.4   5-氧亚基-5-(2-N-Boc-氨基-乙氨基)戊酸-(齐墩果烷-2, 12-二烯-28-基)酯(OA-6c)的合成

  按照合成OA-6a的方法由化合物将OA-6 (200 mg, 0.37 mmol)、N-Boc乙二胺(70.5 mg, 0.44 mmol)、DMAP (53.8 mg, 0.44 mmol)和EDCI (354.6 mg, 1.85 mmol)反应得到了227.8 mg白色固体产物OA-6c, 收率90%. m.p. 79~80 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.24 (s, 1H, CONH), 5.42~5.35 (m, 2H, H-2 & H-3), 5.23 (s, 1H, H-12), 4.98 (s, 1H, NHBoc), 4.04 (d, J＝－11.0 Hz, 1H, CHO), 3.72 (d, J＝－11.0 Hz, 1H, CHO), 3.36~3.25 (m, 4H, HNCH₂CH₂NH), 2.38~2.20 (m, 4H, COCH₂CH₂- CH₂CO), 2.04 (d, J＝10.4 Hz, 1H, H-18), 1.43 (s, 9H, C(CH₃)₃), 1.16, 0.97, 0.95, 0.89, 0.88, 0.87 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.73, 172.29, 156.36, 142.91, 137.45, 122.66, 120.90, 79.17, 70.23, 51.54, 45.76, 45.58, 42.21, 41.32, 40.40, 40.13, 39.87, 39.39, 35.61, 35.43, 35.02, 33.96, 33.50, 33.01, 32.61, 31.47, 31.28, 30.96, 30.37, 27.86, 25.31, 25.00, 23.05, 22.97, 22.29, 21.86, 20.48, 19.00, 15.97, 15.22; IR (KBr) ν: 3426, 3005, 2953, 2925, 2867, 1718, 1646 cm^－1; HRMS calcd for C₄₂H₆₉N₂O₅ [M+H]⁺ 681.5206, found 681.5209.

  3.2.5   5-氧亚基-5-(2-氨基-乙氨基)戊酸-(齐墩果烷- 2, 12-二烯-28-基)酯(OA-6d)的合成

  将OA-6c (100 mg, 0.15 mmol)溶于CH₂Cl₂中, 冷却到0 ℃后加入三氟乙酸(0.33 mL, 4.5 mmol), 再将温度升至室温搅拌2 h, 反应结束后用NH₃•H₂O中和至微碱性(pH＝9, 冰水浴), 反应完毕后用H₂O和CH₂Cl₂对反应混合物进行萃取, 有机层用无水MgSO₄进行干燥, 过滤, 经减压蒸馏的产物, 再经柱层析得到80 mg白色固体产物OA-6d, 收率94%. m.p. 67~68 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.11 (s, 1H, CONH), 5.44~5.37 (m, 2H, H-2 & H-3), 5.24 (s, 1H, H-12), 4.06 (d, J＝－11.0 Hz, 1H, CHO), 3.74 (d, J＝－11.0 Hz, 1H, CHO), 3.32~3.29 (m, 2H, HNCH₂CH₂NH₂), 2.85~2.83 (m, 2H, HNCH₂CH₂NH₂), 2.41~2.24 (m, 6H, COCH₂CH₂CH₂CO & NH₂), 2.06 (d, J＝13.0 Hz, 1H, H-18), 1.18, 0.99, 0.97, 0.90, 0.88 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.80, 171.89, 142.91, 137.45, 122.66, 120.89, 70.23, 51.54, 45.76, 45.58, 42.20, 41.47, 41.32, 40.87, 40.41, 39.39, 35.61, 35.43, 35.10, 33.97, 33.51, 33.01, 32.61, 31.47, 31.29, 30.96, 30.37, 25.32, 25.00, 23.05, 22.97, 22.30, 21.89, 20.60, 19.00, 15.98, 15.22; IR (KBr) ν: 3571, 3433, 3005, 2953, 2925, 2859, 1733, 1653 cm^－1; HRMS calcd for C₃₇H₆₁N₂O₃ [M+H]⁺581.4683, found 581.4667.

  3.2.6   2-(齐墩果烷-2, 12-二烯-28-亚氧基-28-氧)乙酸(OA-9)的合成

  在圆底烧瓶中依次加入OA-8 (1 g, 2.28 mmol)和CH₂Cl₂, 在搅拌下加入(COCl)₂ (0.78 mL, 9.12 mmol)常温下搅拌, 过夜, 反应完毕后减压蒸馏除去溶剂, 再加入乙醇酸(346.8 mg, 4.56 mmol)和四氢呋喃, 在搅拌加入三乙胺(3.16 mL, 22.8 mmol), 室温下反应7 h, 反应完毕后减压蒸馏除去溶剂, 用乙酸乙酯和水对反应混合物进行提取, 并用1 mol/L HCl洗涤至微酸性, 用饱和氯化钠洗涤, 过滤, 柱层析得到420.8 mg白色固体产物OA-9, 收率37%. m.p. 95~96 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.41~5.35 (m, 2H, H-2 & H-3), 5.33 (s, 1H, H-12), 4.66~4.54 (m, 2H, OCH₂CO), 2.88 (dd, J＝4.2, 13.7 Hz, 1H, H-18), 1.15, 0.95, 0.93, 0.90, 0.89, 0.76 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 177.04, 173.17, 143.24, 137.93, 122.88, 121.42, 59.97, 52.06, 46.90, 46.13, 45.93, 41.90, 41.42, 40.76, 39.46, 36.20, 34.44, 33.88, 33.03, 32.20, 31.79, 30.65, 27.58, 25.64, 23.58, 23.31, 23.18, 22.80, 20.37, 19.53, 16.65, 15.56; IR (KBr) ν: 3439, 3001, 2921, 2855, 1733, 1633 cm^－1; HRMS calcd for C₃₂H₄₇O₄ [M－H]^－ 495.3475, found 495.3522.

  3.2.7   齐墩果烷-2, 12-二烯-28-酸-[1-(1-吗啉基)-1-氧亚基]乙酯(OA-9a)的合成

  按照合成OA-6a的方法由化合物将OA-9 (100 mg, 0.20 mmol)、吗啉(0.02 mL, 0.24 mmol)、DMAP (29.3 mg, 0.24 mmol)和EDCI (191.7 mg, 1.00 mmol)反应得到了84.9 mg白色固体产物OA-9a, 收率75%. m.p. 86~87 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.40~5.35 (m, 2H, H-2 & H-3), 5.31 (s, 1H, H-12), 4.71~4.61 (m, 2H, OCH₂CO), 3.68~3.40 (m, 8H, morpholine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.90 (d, J＝10.1 Hz, 1H, H-18), 1.15, 0.95, 0.94, 0.93, 0.90, 0.88, 0.76 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 177.10, 165.47, 143.48, 137.92, 122.68, 121.40, 66.75, 66.41, 60.81, 52.04, 46.98, 46.12, 45.96, 45.13, 41.87, 41.46, 40.72, 39.47, 36.20, 34.43, 33.88, 33.04, 32.36, 31.78, 30.67, 27.62, 25.69, 23.65, 23.30, 23.15, 22.77, 19.52, 16.74, 15.54; IR (KBr) ν: 3415, 2954, 2862, 1734, 1680 cm^－1; HRMS calcd for C₃₆H₅₆NO₄ [M+H]⁺ 566.4210, found 566.4199.

  3.2.8   齐墩果烷-2, 12-二烯-28-酸-[1-(4-甲基-1-哌嗪基)-1-氧亚基]乙酯(OA-9b)的合成

  按照合成OA-6a的方法由化合物将OA-9 (80 mg, 0.16 mmol)、N-甲基哌嗪(0.03 mL, 0.19 mmol)、DMAP (23.2 mg, 0.19 mmol)和EDCI (153.4 mg, 0.80 mmol)反应得到了73.1 mg白色固体产物OA-9b, 收率78%. m.p. 79~80 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.40~5.35 (m, 2H, H-2 & H-3), 5.31 (s, 1H, H-12), 4.72~4.62 (m, 2H, OCH₂CO), 3.62~2.40 (m, 8H, piperazine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.90 (d, J＝9.4 Hz, 1H, H-18), 2.30 (s, 3H, NCH₃), 1.14, 0.95, 0.93, 0.89, 0.88, 0.76 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 176.61, 164.72, 143.04, 137.45, 122.17, 120.92, 60.46, 54.36, 53.98, 51.57, 46.49, 45.65, 45.46, 44.00, 41.39, 40.98, 40.25, 38.99, 35.72, 33.95, 33.41, 32.57, 31.88, 31.67, 31.30, 30.19, 27.16, 25.21, 23.19, 22.82, 22.65, 22.29, 19.05, 16.26, 15.05; IR (KBr) ν: 3008, 2947, 2869, 2796, 1734, 1680 cm^－1; HRMS calcd for C₃₇H₅₉N₂O₃ [M+H]⁺ 579.4526, found 579.4512.

  3.2.9   齐墩果烷-2, 12-二烯-28-酸-[1-(2-N-Boc-氨基-乙氨基)-1-氧亚基]乙酯(OA-9c)的合成

  按照合成OA-6a的方法由化合物将OA-9 (200 mg, 0.40 mmol)、N-Boc乙二胺(76.9 mg, 0.48 mmol)、DMAP (58.6 mg, 0.48 mmol)和EDCI (383.4 mg, 2.00 mmol)反应得到了97.6 mg白色固体产物OA-9c, 收率38%, m.p. 89~91 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.70 (s, 1H, CONH), 5.40~5.36 (m, 3H, H-2 & H-3 & H-12), 4.91 (s, 1H, NHBoc), 4.71~4.66 (m, 1H, OCH), 4.38~4.32 (m, 1H, OCH), 3.53~3.27 (m, 4H, HNCH₂CH₂NH), 2.91 (d, J＝11.2 Hz, 1H, H-18), 1.45 (s, 9H, C(CH₃)₃), 1.18, 0.96, 0.94, 0.93, 0.90, 0.75, 0.74 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 175.77, 167.57, 137.47, 137.43, 122.16, 120.79, 62.37, 62.31, 51.56, 46.58, 45.57, 45.33, 41.50, 41.18, 39.55, 38.94, 38.30, 35.70, 33.95, 33.32, 32.51, 31.85, 31.61, 31.28, 30.17, 27.85, 27.08, 25.26, 23.13, 23.01, 22.81, 22.27, 20.58, 18.97, 16.49, 16.19, 15.01; IR (KBr) ν: 3433, 3426, 3005, 2976, 2925, 2881, 1726, 1690 cm^－1; HRMS calcd for C₃₉H₆₂N₂O₅Na [M+Na]⁺ 661.4557, found 661.4564.

  3.2.10   齐墩果烷-2, 12-二烯-28-酸-[1-(2-氨基-乙氨基)-1-氧亚基]乙酯(OA-9d)的合成

  按照合成OA-6d的方法由化合物将OA-9c (50 mg, 0.08 mmol)和三氟乙酸(0.18 mL, 2.40 mmol)反应得到了42.0 mg白色固体产物OA-9d, 收率99%. m.p. 69~70 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.68 (s, 1H, CONH), 5.40~5.29 (m, 3H, H-2 & H-3 & H-12), 4.66~4.35 (m, 2H, OCH₂CO), 3.43~3.30 (m, 2H, HNCH₂- CH₂NH₂), 2.89~2.87 (m, 2H, HNCH₂CH₂NH₂), 2.26 (d, J＝10.8 Hz, 1H, H-18), 1.16, 0.95, 0.94, 0.92, 0.89, 0.74 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 175.78, 167.22, 138.41, 137.47, 122.13, 120.74, 62.44, 51.52, 46.57, 45.54, 45.30, 41.45, 41.15, 39.11, 38.92, 35.67, 35.61, 33.92, 33.27, 32.47, 31.94, 31.55, 31.25, 30.14, 27.04, 25.24, 23.05, 22.79, 22.71, 22.24, 20.55, 18.94, 16.14, 14.98; IR (KBr) ν: 3462, 3441, 3005, 2953, 2925, 2852, 1733, 1631 cm^－1; HRMS calcd for C₃₄H₅₅N₂O₃ [M+H]⁺ 539.4213, found 539.4201.

  3.3   熊果酸衍生物的合成

  3β-羟基熊果-12-烯-28-羧酸甲酯(UA-1)、3β-甲基磺酰氧基熊果-12-烯-28-酸甲酯(UA-2)、熊果烷-2, 12-二烯-28-羧酸甲酯(UA-3)、熊果烷-2, 12-二烯-28-醇(UA-4)的合成参考文献[15], 均得到白色固体产物; 3β-甲基磺酰氧基熊果-12-烯-28-酸(UA-7)、熊果烷-2, 12-二烯-28-酸(UA-8)的合成参考文献[16], 均得到淡黄色固体产物.

  3.3.1   5-氧亚基-5-(熊果烷-2, 12-二烯-28-基氧基)戊酸(UA-6)的合成

  按照OA-6的合成方法合成白色固体UA-6, 收率71%. m.p. 59~60 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.44~5.36 (m, 2H, H-2 & H-3), 5.18 (s, 1H, H-12), 4.09 (d, J＝－11.0 Hz, 1H, CHO), 3.68 (d, J＝－11.0 Hz, 1H, CHO), 2.44~2.39 (m, 4H, COCH₂CH₂CH₂CO), 1.11, 1.02, 0.97, 0.96, 0.93, 0.90, 0.82 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 178.34, 172.91, 137.97, 137.89, 125.93, 121.43, 71.26, 54.49, 52.05, 46.15, 42.12, 41.13, 40.08, 39.33, 37.00, 36.05, 35.71, 34.47, 33.30, 32.94, 32.21, 31.81, 30.53, 25.94, 23.52, 23.38, 23.25, 22.78, 21.23, 19.97, 19.45, 17.25, 16.56, 15.91; IR (KBr) ν: 3446, 3008, 2953, 2867, 1740, 1713 cm^－1; HRMS calcd for C₃₅H₅₃O₄ [M－H]^－ 537.3944, found 537.3952.

  3.3.2   5-氧亚基-5-(1-吗啉基)戊酸-(熊果烷-2, 12-二烯-28-基)酯(UA-6a)的合成

  按照OA-6a的合成方法合成白色固体UA-6a, 收率86%, m.p. 51~52 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.45~5.37 (m, 2H, H-2 & H-3), 5.18 (s, 1H, H-12), 4.10 (d, J＝－11.0 Hz, 1H, CHO), 3.67~3.47 (m, 9H, CHO & morpholine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.44~2.37 (m, 4H, COCH₂CH₂CH₂CO), 1.12, 1.03, 0.98, 0.97, 0.94, 0.91, 0.83 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.86, 170.41, 137.53, 137.40, 125.41, 120.91, 70.67, 66.42, 66.16, 54.04, 51.56, 45.65, 45.46, 41.65, 41.43, 40.64, 39.59, 38.86, 36.52, 35.56, 35.26, 33.98, 33.00, 31.75, 31.57, 31.31, 30.04, 25.47, 23.01, 22.76, 22.29, 20.72, 20.00, 18.95, 16.76, 16.10, 15.42; IR (KBr) ν: 2961, 2917, 2859, 1733, 1661 cm^－1; HRMS calcd for C₃₉H₆₂NO₄ [M+H]⁺ 608.4680, found 608.4664.

  3.3.3   5-氧亚基-5-(4-甲基-1-哌嗪基)戊酸-(熊果烷- 2, 12-二烯-28-基)酯(UA-6b)的合成

  按照OA-6b的合成方法合成白色固体UA-6b, 收率63%. m.p. 44~45 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.41~5.35 (m, 2H, H-2 & H-3), 5.18~5.16 (m, 1H, H-12), 4.08 (d, J＝－11.0 Hz, 1H, CHO), 3.65 (d, J＝ －11.0 Hz, 1H, CHO), 3.63~2.35 (m, 12H, COCH₂CH₂- CH₂CO & piperazine H-2×2 & H-3×2 & H-5×2 & H-6×2), 1.10, 1.01, 0.96, 0.92, 0.89 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.85, 170.10, 137.50, 137.36, 125.37, 120.89, 70.60, 54.67, 54.21, 54.01, 51.53, 45.62, 45.46, 44.86, 41.61, 40.95, 40.61, 39.56, 38.83, 36.48, 35.53, 35.23, 33.95, 33.08, 31.71, 31.28, 30.01, 25.45, 22.97, 22.87, 22.73, 22.26, 20.69, 20.08, 18.93, 16.73, 16.07, 15.39; IR (KBr) ν: 3005, 2925, 2859, 2794, 1733, 1653 cm^－1; HRMS calcd for C₄₀H₆₅N₂O₃ [M+H]⁺ 621.4996, found 621.4980.

  3.3.4   5-氧亚基-5-(2-N-Boc-氨基-乙氨基)戊酸-(熊果烷-2, 12-二烯-28-基)酯(UA-6c)的合成

  按照OA-6c的合成方法合成白色固体UA-6c, 收率77%. m.p. 73~74 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.21 (s, 1H, CONH), 5.43~5.35 (m, 2H, H-2 & H-3), 5.17 (s, 1H, H-12), 4.96 (s, 1H, NHBoc), 4.08 (d, J＝－11.0 Hz, 1H, CHO), 3.64 (d, J＝－11.0 Hz, 1H, CHO), 3.35~3.25 (m, 4H, HNCH₂CH₂NH), 2.37~2.20 (m, 4H, COCH₂CH₂CH₂CO), 1.43 [s, 9H, C(CH₃)₃], 1.10, 1.01, 0.96, 0.95, 0.92, 0.89, 0.82 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.74, 172.31, 137.52, 137.40, 125.58, 125.41, 120.93, 79.16, 70.70, 54.06, 51.56, 45.66, 44.68, 41.64, 40.64, 40.11, 39.86, 39.59, 38.85, 36.52, 35.56, 35.24, 35.03, 33.98, 33.01, 31.73, 31.31, 30.03, 27.86, 25.46, 22.98, 22.89, 22.76, 22.29, 20.72, 20.50, 18.96, 16.76, 16.08, 15.41; IR (KBr) ν: 3571, 3441, 3005, 2953, 2925, 2859, 1726, 1638 cm^－1; HRMS calcd for C₄₂H₆₉N₂O₅ [M+H]⁺ 681.5206, found 681.5205.

  3.3.5   5-氧亚基-5-(2-氨基-乙氨基)戊酸-(熊果烷- 2, 12-二烯-28-基)酯(UA-6d)的合成

  按照OA-6c的合成方法合成白色固体UA-6c, 收率88%. m.p. 62~63 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.12 (s, 1H, CONH), 5.43~5.35 (m, 2H, H-2 & H-3), 5.17 (s, 1H, H-12), 4.08 (d, J＝－11.0 Hz, 1H, CHO), 3.65 (d, J＝－11.0 Hz, 1H, CHO), 3.30 (d, J＝5.4 Hz, 2H, HNCH₂CH₂NH₂), 2.83 (d, J＝5.0 Hz, 2H, HNCH₂- CH₂NH₂), 2.38~2.22 (m, 4H, COCH₂CH₂CH₂CO), 1.10, 1.01, 0.96, 0.95, 0.92, 0.89, 0.81 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 172.81, 171.93, 137.52, 137.40, 125.41, 120.92, 70.70, 54.05, 51.56, 45.65, 41.64, 41.44, 40.87, 40.64, 39.59, 38.86, 36.52, 35.56, 35.25, 35.10, 33.98, 33.01, 31.73, 31.31, 30.03, 25.46, 23.00, 22.89, 22.76, 22.29, 20.72, 20.62, 18.96, 16.76, 16.09, 15.42; IR (KBr) ν: 3441, 3005, 2961, 2925, 2859, 1733, 1646 cm^－1; HRMS calcd for C₃₇H₆₁N₂O₃ [M+H]⁺ 581.4683, found 581.4668.

  3.3.6   2-(熊果烷-2, 12-二烯-28-亚氧基-28-氧)乙酸(UA-9)的合成

  按照OA-9的合成方法合成白色固体UA-9, 收率40%. m.p. 103~104 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.43~5.36 (m, 2H, H-2 & H-3), 5.30 (s, 1H, H-12), 4.62~4.54 (m, 2H, OCH₂CO), 2.27 (d, J＝11.2 Hz, 1H, H-18), 1.10, 0.95, 0.89, 0.88, 0.86, 0.77 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 176.77, 173.12, 137.88, 137.70, 126.13, 121.44, 59.91, 52.95, 52.07, 48.31, 46.06, 42.22, 40.98, 39.66, 39.17, 38.79, 36.42, 36.13, 34.45, 32.49, 31.81, 30.67, 27.94, 24.29, 23.37, 23.27, 22.78, 21.12, 19.48, 16.97, 16.84, 15.72; IR (KBr) ν: 3446, 2947, 2928, 2869, 1734, 1640 cm^－1; HRMS calcd for C₃₂H₄₇O₄ [M－H]^－ 495.3553, found 495.3519.

  3.3.7   熊果烷-2, 12-二烯-28-酸-[1-(1-吗啉基)-1-氧亚基]乙酯(UA-9a)的合成

  按照OA-9a的合成方法合成白色固体UA-9a, 收率64%. m.p. 112~113 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.40~5.35 (m, 2H, H-2 & H-3), 5.27 (s, 1H, H-12), 4.72~4.59 (m, 2H, OCH₂CO), 3.67~3.40 (m, 8H, morpholine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.28 (d, J＝11.2 Hz, 1H, H-18), 1.09, 0.94, 0.88, 0.86, 0.85, 0.77 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 176.89, 165.50, 137.91, 137.86, 125.90, 121.42, 66.75, 66.41, 60.75, 52.95, 52.04, 48.39, 46.02, 45.14, 42.19, 40.94, 39.66, 39.16, 38.77, 36.59, 36.13, 34.44, 32.43, 31.80, 30.70, 27.97, 24.29, 23.40, 23.25, 22.77, 21.15, 19.48, 16.99, 15.71; IR (KBr) ν: 3014, 2954, 2928, 2869, 1739, 1673 cm^－1; HRMS calcd for C₃₆H₅₆NO₄ [M+H]⁺ 566.4210, found 566.4201.

  3.3.8   熊果烷-2, 12-二烯-28-酸-[1-(4-甲基-1-哌嗪基)-1-氧亚基]乙酯(UA-9b)的合成

  按照OA-9b的合成方法合成白色固体UA-9b, 收率73%. m.p. 92~93 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 5.40~5.34 (m, 2H, H-2 & H-3), 5.27 (s, 1H, H-12), 4.72~4.60 (m, 2H, OCH₂CO), 3.61~2.39 (m, 8H, piperazine H-2×2 & H-3×2 & H-5×2 & H-6×2), 2.29 (s, 3H, NCH₃), 2.27 (s, 1H, H-18), 1.06, 0.94, 0.88, 0.86, 0.85, 0.77 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 176.39, 164.75, 137.46, 137.39, 125.40, 120.94, 60.41, 54.38, 53.99, 52.46, 51.58, 47.90, 45.48, 44.03, 41.71, 40.47, 39.19, 38.69, 38.29, 36.10, 35.66, 33.96, 31.96, 31.32, 30.23, 27.50, 23.80, 22.92, 22.78, 22.28, 20.66, 19.00, 16.50, 16.45, 15.22; IR (KBr) ν: 3008, 2928, 2869, 2796, 1734, 1680 cm^－1; HRMS calcd for C₃₇H₅₉N₂O₃ [M+H]⁺ 579.4526, found 579.4512.

  3.3.9   熊果烷-2, 12-二烯-28-酸-[1-(2-N-Boc-氨基-乙氨基)-1-氧亚基]乙酯(UA-9c)的合成

  按照OA-9c的合成方法合成白色固体UA-9c, 收率73%. m.p. 82~83 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.70 (s, 1H, CONH), 5.44~5.39 (m, 2H, H-2 & H-3), 5.37 (s, 1H, H-12), 4.90 (s, 1H, NHBoc), 4.71~4.67 (m, 1H, OCH), 4.39~4.32 (m, 1H, OCH), 3.54~3.27 (m, 4H, HNCH₂CH₂NH), 2.91 (d, J＝11.7 Hz, 1H, H-18), 1.45 (s, 9H, C(CH₃)₃), 1.18, 1.12, 0.97, 0.93, 0.90, 0.75, 0.74 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 175.78, 167.59, 156.03, 137.47, 122.15, 120.80, 79.14, 62.36, 52.53, 51.54, 47.99, 46.58, 45.56, 41.49, 41.16, 40.17, 38.93, 38.29, 35.70, 33.95, 33.31, 32.52, 31.84, 31.60, 31.29, 30.17, 27.85, 25.27, 23.13, 22.80, 22.77, 22.28, 20.59, 18.97, 16.50, 16.19, 15.02; IR (KBr) ν: 3441, 3419, 3005, 2953, 2932, 2873, 1733, 1690 cm^－1; HRMS calcd for C₃₉H₆₂N₂O₅Na [M+Na]⁺ 661.4557, found 661.4558.

  3.3.10   熊果烷-2, 12-二烯-28-酸-[1-(2-氨基-乙氨基)-1-氧亚基]乙酯(UA-9d)的合成

  按照OA-9d的合成方法合成白色固体UA-9d, 收率95%. m.p. 83~84 ℃; ¹H NMR (500 MHz, CDCl₃) δ: 6.65 (s, 1H, CONH), 5.43~5.29 (m, 3H, H-2 & H-3 & H-12), 4.66~4.60 (m, 1H, OCH), 4.46~4.36 (m, 1H, OCH), 3.42~3.28 (m, 2H, HNCH₂CH₂NH₂), 2.90~2.87 (m, 2H, HNCH₂CH₂NH₂), 2.26 (d, J＝10.8 Hz, 1H, H-18), 1.16, 0.95, 0.94, 0.92, 0.89, 0.74 (each s, each 3H); ¹³C NMR (125 MHz, CDCl₃) δ: 175.74, 167.15, 143.63, 137.46, 122.13, 120.72, 62.44, 51.52, 46.56, 45.54, 45.29, 41.44, 41.15, 41.06, 40.67, 40.16, 38.92, 35.67, 33.91, 33.27, 32.46, 31.94, 31.55, 31.25, 30.13, 27.04, 25.24, 23.04, 22.79, 22.24, 20.54, 18.94, 16.14, 14.98; IR(KBr) ν: 3456, 3426, 3005, 2953, 2932, 2867, 1740, 1682 cm^－1; HRMS calcd for C₃₄H₅₅N₂O₃ [M+H]⁺ 539.4213, found 539.4202.

  3.4   体外抗肿瘤细胞增殖测试

  3.4.1   细胞培养

  人肺腺癌细胞A549、人乳腺癌MCF-7细胞株和人肝肿瘤细胞HepG2细胞株的培养基为RPMI-1640, 其中含有10%(体积分数)胎牛血清、0.1 mg/mL链霉素、100 U/mL青霉素.细胞在37 ℃、5% CO₂培养箱内进行培养.

  3.4.2   抗肿瘤细胞增殖实验

  将适当浓度的肿瘤细胞接种到96孔板上, 置于含有5%二氧化碳37 ℃恒温培养箱中培养24 h.然后在各孔中加入20 μL含有不同浓度的测试药物, 药物的浓度范围为0.0001~100 μmol/L, 加入药物后继续培养72 h.然后在各孔中加入20 μL浓度为5%的3-(4, 5-二甲基-2-噻唑)-2, 5-二苯基溴化四氮唑噻唑蓝(MTT), 再培养4 h, 然后倒去培养板上的液体, 在室温下晾干培养板.在彻底干燥后, 在各孔中加入200 μL酸性异丙醇(每500 mL异丙醇中加有50 μL的盐酸)来溶解所形成的MTT甲瓒化产物, 通过测定570 nm的每个孔的光密度来确定甲瓒产物的量.通过MTT法测定并计算出各化合物的IC₅₀值.

  3.5   UA-9d体外抗缺氧诱导因子1α (HIF-1α)活性

  通过缺氧反应原件(HRE)依赖性分子测定法测定UA-9d抑制缺氧诱导因子表达的能力.在细胞长到培养瓶的50%~80%的时候, 将含有6个拷贝基因的pGL3- HRE质粒载体转染到Hep3B细胞中, 37 ℃ CO₂培养箱中培养24 h; 将转染后的细胞用不同浓度(分别为0, 0.1, 0.3, 1, 3, 10 μmol/L)的UA-9d处理, 低氧条件下孵育16 h, 进行荧光素酶测定.

  辅助材料(Supporting Information)化合物OA-6, OA-6a~OA-6d, UA-6, UA-6a~UA-6d, OA-9, OA-9a~OA-9d, UA-9, UA-9a~UA-9d的¹H NMR、¹³C NMR和HRMS谱图.这些材料可以免费从本刊网站(http://sioc-journal.cn/)上下载.

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• 

  图 1  齐墩果酸和熊果酸的结构

  Figure 1  Structure of oleanolic and ursolic acid

  下载: 全尺寸图片 幻灯片
  

  图 2  CDDO、CDDO-Me和CDDO-Im的结构

  Figure 2  Structures of CDDO, CDDO-Me and CDDO-Im

  下载: 全尺寸图片 幻灯片
  

  图 3  OA-5结构及局部¹H NMR图

  Figure 3  Structure and ¹H NMR partially view of OA-5

  下载: 全尺寸图片 幻灯片
  

  图式 1  齐墩果酸衍生物OA-6~OA-6d和熊果酸衍生物UA-6~UA-6d的合成

  Scheme 1  Synthesis of OA derivatives of OA-6~OA-6d and UA derivatives of UA-6~UA-6d

  下载: 全尺寸图片 幻灯片
  

  图式 2  齐墩果酸衍生物OA-9~OA-9d和熊果酸衍生物UA-9~UA-9d的合成

  Scheme 2  Synthesis of OA derivatives of OA-6~OA-6d and UA derivatives of UA-9~UA-9d

  下载: 全尺寸图片 幻灯片
  

  图 4  UA-9d抑制HIF-1α的表达情况

  Figure 4  UA-9d inhibits the expression of hypoxia-inducible factor 1α

  (－): negative control, under the condition of 20% O₂, dissolve in DMSO; (+): positive control, under the condition of 1% O₂, dissolve in DMSO. **P < 0.01.

  下载: 全尺寸图片 幻灯片

  表 1  OA和UA衍生物的抑制A549、MCF-7和HepG2细胞的IC₅₀值

  Table 1.  IC₅₀ of OA and UA derivatives on A549, MCF-7 and HepG2

  Sample	MW	IC50/(μmol•L－1)
  		A549	MCF-7	HepG2
  顺铂		2.3±0.19	2.6±0.94	4.7±1.3
  紫杉醇	853.92	96.6±10.4	89.7±6.59	108.6±9.7
  OA	456.70	＞50	＞50	＞50
  OA-6	538.80	9.60±1.87	11.66±1.76	21.24±2.12
  OA-6a	607.91	＞50	＞50	＞50
  OA-6b	619.96	＞50	＞50	42.00±1.58
  OA-6c	681.00	＞50	＞50	＞50
  OA-6d	580.88	39.15±2.95	13.54±1.32	4.87±1.65
  OA-9	496.72	24.81±1.88	34.04±2.01	41.62±4.36
  OA-9a	565.83	＞50	＞50	＞50
  OA-9b	578.87	＞50	36.18±5.52	20.55±1.65
  OA-9c	638.92	＞50	＞50	＞50
  OA-9d	538.80	15.70±3.16	5.25±0.71	4.98±1.22
  UA	456.70	5.81±0.32	9.57±2.55	17.78±1.63
  UA-6	538.80	15.43±2.79	13.79±1.36	23.44±2.14
  UA-6a	607.91	＞50	＞50	＞50
  UA-6b	619.96	＞50	＞50	42.00±3.24
  UA-6c	681.00	＞50	＞50	＞50
  UA-6d	580.88	37.39±12.17	9.16±0.17	8.88±2.50
  UA-9	496.72	14.50±2.01	19.81±1.03	24.74±1.52
  UA-9a	565.83	＞50	＞50	＞50
  UA-9b	578.87	10.05±1.40	12.14±2.05	17.94±1.66
  UA-9c	638.92	＞50	＞50	＞50
  UA-9d	538.80	5.61±3.16	3.52±0.71	4.51±1.22
  a The unit of IC50 value of Taxol is nmol•L－1.

  下载: 导出CSV
•