11-羰基-20-乙氧基济源冬凌草甲素及其14-OH酯化系列衍生物和用途

11-羰基-20-乙氧基济源冬凌草甲素及其14-oh酯化系列衍生物和用途
技术领域
1.本发明涉及天然产物及药物化学领域，具体涉及济源冬凌草甲素类化合物：11-羰基-20-乙氧基济源冬凌草甲素14-oh酯化系列衍生物、其合成方法及应用。


背景技术：

2.冬凌草，唇形科香茶菜属植物，味苦、甘，性微寒。具有清热解毒，活血止痛的功效。冬凌草的化学成分研究主要涉及二萜类、挥发油、黄酮和有机酸等，其中冬凌草甲素是冬凌草中主要的抗癌有效成分，是一种无色棱柱状结晶性粉末，几乎不溶于水，微溶于乙醚、甲醇和乙醇等有机溶剂。冬凌草甲素在体外可抑制多种肿瘤细胞生长，包括胃癌、食管癌、鼻咽癌、肝癌、肺癌、膀胱癌、结肠癌、白血病和宫颈癌等癌细胞。本发明人前期通过一系列提取分离方法从济源冬凌草中得到骨架为对映-贝壳杉烯型二萜的先导化合物：济源冬凌草甲素(joa)，是一个醛式与半缩醛式的混合物，主要以半缩醛式形式存在，使其稳定性不佳，影响了joa的存储和成药性，发明人考虑直接将joa的11位和20位氧化后得到11-羰基-20-乙氧基济源冬凌草甲素(joa11)。初步的抗肿瘤活性测试表明joa11本身具有较好的抗肿瘤活性。
3.此类化合物化学结构上的不同取代基对药理活性有着至关重要的影响。因此对joa11的结构修饰希望得到有良好抗肿瘤活性、高生物利用度、理化性质稳定且低毒的化合物，并能尽早开发成新药，有利于我国自主知识产权的新药研发。相关内容目前尚未见相关文献报道。


技术实现要素：

4.本发明目的在于提供11-羰基-20-乙氧基济源冬凌草甲素及其一系列14-oh酯化系列衍生物，及其光学活性体或消旋体，非对映异构体混合物或药学可接受的盐，并提高其抗肿瘤作用及稳定性，为其在临床上的应用提供可能。
5.本发明的另一目的在于提供其制备方法以及其在制备抗肿瘤药物的应用。
6.为实现本发明目的，本发明将11-羰基-20-乙氧基济源冬凌草甲素与有机酸酯化得到系列衍生物，提高其稳定性，同时保留或增强其抗肿瘤活性。本发明所述11-羰基-20-乙氧基济源冬凌草甲素结构通式如下所示：本发明提供的11-羰基-20-乙氧基济源冬凌草甲素酯化衍生物结构通式如下所示：
r为与11-羰基-20-乙氧基济源冬凌草甲素14位羟基酯化后的有机酸基团；所述有机酸选自：被氯甲基、卤素、硝基单取代的苯甲酸或被双取代的苯甲酸；被叔丁氧羰基单取代的丙氨酸、苯丙氨酸、缬氨酸、亮氨酸、5-氨基戊酸、6-氨基己酸、2-氨基丁酸、γ-氨基丁酸、吡咯烷甲酸、环丁烷甲酸；被卤素、甲基单取代的烟酸；乙酸、丙酸、丙烯酸、丁酸、2-乙基丁酸、环丙乙酸、嘧啶甲酸、2-甲基5-嘧啶甲酸、5-溴-2-嘧啶甲酸、吡嗪甲酸、糠酸、吲哚甲酸；肉桂酸或被甲基、甲氧基、硝基单取代的肉桂酸；4-吡啶丙烯酸、3-吡啶丙烯酸、2-吡啶丙烯酸；被三氟乙酸成盐的丙氨酸、苯丙氨酸、5-氨基戊酸、6-氨基己酸、2-氨基丁酸、吡咯烷甲酸、环丁烷甲酸。进一步，所述有机酸选自：4-氯甲基苯甲酸、5-氯-2硝基苯甲酸、被溴、氟单取代的苯甲酸；被叔丁氧羰基单取代的丙氨酸、苯丙氨酸、缬氨酸、亮氨酸、5-氨基戊酸、6-氨基己酸、2-氨基丁酸、γ-氨基丁酸、吡咯烷甲酸、环丁烷甲酸；被甲基或氟单取代的烟酸；乙酸、丙酸、丙烯酸、丁酸、2-乙基丁酸、环丙乙酸、4-嘧啶甲酸、2-甲基5-嘧啶甲酸、5-溴-2-嘧啶甲酸、2-吡嗪甲酸、糠酸、3-吲哚甲酸；肉桂酸或被甲基、甲氧基、硝基单取代的肉桂酸；4-吡啶丙烯酸、3-吡啶丙烯酸、2-吡啶丙烯酸；被三氟乙酸成盐的丙氨酸、苯丙氨酸、5-氨基戊酸、6-氨基己酸、2-氨基丁酸、吡咯烷甲酸、环丁烷甲酸。
7.本发明所述的11-羰基-20-乙氧基济源冬凌草甲素酯化衍生物通过如下合成路线得到：
8.1、称取20-乙氧基济源冬凌草甲素使其完全溶解于二氯甲烷中，冰浴条件下加入戴斯马丁氧化剂，反应结束后饱和硫代硫酸钠溶液猝灭，萃取干燥后柱色谱纯化得到目标化合物(joa11)。
9.2、将上述所得母核11-羰基-20-乙氧基济源冬凌草甲素溶于二氯甲烷中，冰浴条件下加入相应的有机酸，再依次加入催化剂1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(edci)和4-二甲氨基吡啶(dmap)反应。反结束后萃取干燥，浓缩，柱色谱纯化得到目标衍生物。
10.3、将上述所得母核11-羰基-20-乙氧基济源冬凌草甲素14-位酯化衍生物溶于二氯甲烷中，加入过量的三氟乙酸使其脱掉boc保护基并成盐，将体系旋蒸浓缩后加入异丙醚使其析出得到成盐的目标衍生物。
11.本发明创新点及优点：本发明选用济源冬凌草甲素(joa)作为母核，经过氧化、酯化过程，得到所需的衍生物，该类化合物具有抗癌活性，可用于制备抗肿瘤药物，应用于临床治疗食管癌、胰腺癌、肺癌、宫颈癌、结肠癌、乳腺癌和前列腺癌等，具有很好的开发前景。
具体实施方式
12.通过以下具体实例进一步举例说明本发明，但应注意本发明的范围并不接受这些实施例的任何限制。
13.实施例1：
[0014][0015]
称取400mg 20-乙氧基济源冬凌草甲素使其完全溶解于二氯甲烷中，冰浴条件下加入676mg戴斯马丁氧化剂，反应结束后饱和硫代硫酸钠溶液猝灭，萃取干燥后柱色谱纯化得白色固体，产率为70％-75％。1h nmr(400mhz,dmso-d6)δ6.00(s,1h),5.63(d,j＝1.2hz,1h),5.56(d,j＝2.8hz,1h),4.98(s,1h),4.72(d,j＝2.9hz,1h),4.21
–
4.18(m,1h),3.82
–
3.77(m,1h),3.41
–
3.36(m,1h),2.97(ddd,j＝5.2,2.4,1.2hz,1h),2.75(dd,j＝18.6,5.1hz,1h),2.46(dt,j＝14.7,2.3hz,1h),2.38(dd,j＝18.4,2.3hz,1h),2.03(dt,j＝13.4,2.3hz,1h),1.74(s,1h),1.63(dt,j＝13.9,5.1hz,1h),1.52
–
1.43(m,1h),1.40(s,1h),1.39
–
1.37(m,2h),1.35(d,j＝5.5hz,1h),1.09(t,j＝7.0hz,4h),0.89(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ204.30,202.90,148.43,120.64,97.48,71.89,65.17,61.80,57.41,57.01,47.47,46.53,42.67,41.22,33.88,31.94,30.10,23.50,20.61,18.12,14.85.
[0016]
实施例2
[0017][0018]
称取200mg joa11将其完全溶解于10ml的二氯甲烷中，冰浴条件下加入118mg的4-氯甲基苯甲酸，再依次加入催化剂1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(edci)133mg和4-二甲氨基吡啶(dmap)8mg。反应结束后萃取干燥，浓缩，柱色谱纯化得白色固体产物，产率83％。1h nmr(400mhz,dmso-d6)δ7.89
–
7.85(m,2h),7.61
–
7.58(m,2h),6.31
–
6.29(m,1h),6.16(d,j＝1.5hz,1h),5.78(s,1h),5.02(s,1h),4.83(s,2h),4.20
–
4.16(m,1h),3.84(dq,j＝9.4,7.1hz,1h),3.44
–
3.39(m,1h),2.93(dd,j＝18.7,5.2hz,1h),2.60
–
2.53(m,1h),2.49(d,j＝8.9hz,1h),2.11(s,1h),2.01(dq,j＝14.1,2.6hz,1h),1.69(dt,j＝
14.0,5.1hz,1h),1.60
–
1.52(m,1h),1.48(dd,j＝11.7,5.3hz,1h),1.42(s,2h),1.39(s,1h),1.35
–
1.22(m,1h),1.19(t,j＝7.0hz,3h),1.15
–
1.08(m,1h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.85,201.10,164.19,146.24,143.34,129.48(*2),129.22(*2),128.76,122.55,97.57,76.61,65.26,62.04,57.26,55.95,47.20,46.55,45.08,42.89,38.75,33.87,31.89,29.93,23.27,20.48,18.07,14.76.
[0019]
实施例3
[0020][0021]
用140mg 5-氯-2-硝基苯甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为53％。1h nmr(400mhz,dmso-d6)δ8.14(d,j＝8.7hz,1h),7.92(dd,j＝8.7,2.3hz,1h),7.87(d,j＝2.3hz,1h),6.36(s,1h),6.13(d,j＝1.4hz,1h),5.81(s,1h),5.05(s,1h),4.11
–
4.08(m,1h),3.95
–
3.89(m,1h),3.50
–
3.44(m,1h),3.44
–
3.41(m,1h),2.93(dd,j＝18.7,5.1hz,1h),2.59(dd,j＝18.5,2.4hz,1h),2.48
–
2.39(m,1h),2.08(s,1h),2.02
–
1.96(m,1h),1.66(dt,j＝14.1,5.1hz,1h),1.55(ddd,j＝13.9,9.5,7.4hz,1h),1.49
–
1.44(m,1h),1.44
–
1.40(m,2h),1.39(s,1h),1.19(t,j＝7.0hz,3h),1.15
–
1.08(m,1h),0.88(s,3h),0.80(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.55,200.54,162.75,145.86,145.60,138.64,132.65,129.16,127.93,126.36,122.76,97.46,78.41,64.90,62.00,57.44,55.81,46.48,42.86,39.79,38.25,33.86,31.84,29.89,23.17,20.52,18.08,14.73.
[0022]
实施例4
[0023][0024]
用140mg 4-溴苯甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为64％。1h nmr(400mhz,dmso-d6)δ7.80
–
7.73(m,4h),6.30(s,1h),6.16(d,j＝1.4hz,1h),5.78(s,1h),5.02(s,1h),4.16(d,j＝4.4hz,1h),3.84(dq,j＝9.4,7.0hz,1h),3.42(dq,j＝9.3,7.0hz,1h),2.92(dd,j＝18.7,5.1hz,1h),2.56(dd,j＝18.5,2.3hz,1h),2.49(d,j＝11.4hz,1h),2.10(s,1h),2.04
–
1.96(m,1h),1.69(dt,j＝14.1,5.1hz,1h),1.55(ddd,j＝14.2,10.2,7.5hz,1h),1.48(dd,j＝11.7,5.2hz,1h),1.42(s,2h),1.40
–
1.37(m,1h),1.35
–
1.21(m,1h),1.18(t,j＝7.0hz,3h),1.12(dd,j＝11.5,6.6hz,1h),0.86(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.76,201.02,163.93,146.18,132.08(*2),131.02(*2),128.15,127.85,122.61,97.57,76.80,65.23,62.07,57.26,55.95,47.20,46.55,42.89,38.71,33.87,31.88,29.92,23.25,20.48,18.07,14.75.
[0025]
实施例5
[0026][0027]
用97mg 4-氟苯甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为64％。1h nmr(400mhz,dmso-d6)δ7.95
–
7.90(m,2h),7.37(t,j＝8.8hz,2h),6.30
–
6.28(m,1h),6.15(d,j＝1.4hz,1h),5.78(s,1h),5.02(s,1h),4.18
–
4.14(m,1h),3.87
–
3.81(m,1h),3.45
–
3.38(m,1h),2.92(dd,j＝18.7,5.1hz,1h),2.56(dd,j＝18.5,2.5hz,1h),2.47(s,1h),2.11(s,1h),2.00(dq,j＝14.5,2.7hz,1h),1.69(dt,j＝14.1,5.1hz,1h),1.60
–
1.52(m,1h),1.48(dd,j＝11.8,5.2hz,1h),1.42(s,2h),1.39(s,1h),1.18(t,j＝7.0hz,4h),1.16
–
1.08(m,1h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.82,201.07,163.63,146.23,132.10,132.00,125.58,122.57,116.22,115.99,97.57,76.64,65.25,62.05,57.24,55.95,47.19,46.53,42.90,38.74,33.88,31.88,29.93,23.26,20.48,18.07,14.75.
[0028]
实施例6
[0029][0030]
用151mg boc-l-缬氨酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为62％。1h nmr(400mhz,dmso-d6)δ7.24(d,j＝7.5hz,1h),6.06(s,1h),6.02(s,1h),5.73(d,j＝10.9hz,1h),4.99(s,1h),4.14(d,j＝4.4hz,1h),3.91(q,j＝7.4hz,1h),3.82
–
3.77(m,1h),3.37(dd,j＝9.2,7.0hz,1h),3.11(d,j＝4.6hz,1h),2.84(dd,j＝18.6,5.2hz,1h),2.49
–
2.45(m,1h),2.01(s,1h),1.97(s,1h),1.69
–
1.63(m,1h),1.55(d,j＝7.5hz,1h),1.53(s,1h),1.52
–
1.50(m,1h),1.42(s,2h),1.39(s,2h),1.36(s,9h),1.28(q,j＝7.4hz,3h),1.15(s,1h),1.13(s,2h),1.11(s,1h),0.88(s,3h),0.82(s,6h).
13
c nmr(101mhz,dmso-d6)δ202.72,200.85,171.72,155.29,146.32,121.90,97.41,78.04,76.06,64.88,61.88,57.44,55.64,54.79,53.50,47.38,46.56,42.80,39.84,38.60,33.84,32.57,31.88,29.91,28.05,27.70,23.22,20.50,18.46,18.08,14.65,13.34.
[0031]
实施例7
[0032][0033]
用184mg boc-l-苯丙氨酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为76％。1h nmr(400mhz,dmso-d6)δ7.37(d,j＝7.7hz,1h),7.26(d,j＝1.5hz,1h),7.25(s,1h),7.22(s,2h),7.21
–
7.20(m,1h),6.06(s,1h),6.03(s,1h),5.69(s,1h),
5.01(s,1h),4.23(d,j＝4.4hz,1h),4.13(q,j＝7.6hz,1h),3.83(dq,j＝9.3,7.0hz,1h),3.44
–
3.37(m,1h),3.06
–
3.01(m,1h),2.89(d,j＝7.8hz,2h),2.86
–
2.79(m,1h),2.46(d,j＝2.3hz,1h),1.98(d,j＝9.0hz,2h),1.67(dt,j＝13.8,4.8hz,1h),1.57
–
1.49(m,1h),1.42(s,2h),1.42
–
1.39(m,2h),1.33(s,9h),1.19(t,j＝7.2hz,1h),1.15(s,1h),1.13(s,2h),1.11(s,1h),0.89(d,j＝3.9hz,3h),0.83(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.82,200.82,171.00,155.10,146.12,137.21,129.02,128.11,126.41,122.12,97.40,78.22,76.22,64.80,61.88,57.46,55.67,55.56,47.31,46.59,42.82,39.82,38.50,36.33,33.85,31.90,29.93,28.02,27.66,23.23,20.70,20.51,18.08,14.69,14.04.
[0034]
实施例8
[0035][0036]
用131mg boc-l-丙氨酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为71％。1h nmr(400mhz,dmso-d6)δ7.31(d,j＝7.2hz,1h),6.06(s,1h),6.01(s,1h),5.72(s,1h),4.99(s,1h),4.13(d,j＝4.5hz,1h),3.97(t,j＝7.2hz,1h),3.82
–
3.76(m,1h),3.37(dd,j＝9.5,7.2hz,1h),3.10(d,j＝4.7hz,1h),2.83(dd,j＝18.6,5.1hz,1h),2.45(d,j＝13.2hz,1h),1.99(s,1h),1.65(dt,j＝14.1,5.0hz,1h),1.55
–
1.48(m,1h),1.45(d,j＝5.3hz,1h),1.42(d,j＝4.8hz,2h),1.39(s,2h),1.35(s,9h),1.23(dd,j＝8.6,6.6hz,1h),1.17(d,j＝7.3hz,3h),1.14(s,1h),1.12(s,2h),1.11(s,1h),0.88(s,3h),0.81(s,3h).
13
cnmr(101mhz,dmso-d6)δ202.89,200.87,172.15,155.01,146.26,122.08,97.39,78.08,76.03,64.82,61.88,57.36,55.66,49.13,47.24,46.56,42.81,39.80,38.57,33.86,31.89,29.91,28.08,27.70,23.20,20.51,18.06,16.67,14.70.
[0037]
实施例9
[0038][0039]
用161mg boc-l-亮氨酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为39％。1h nmr(400mhz,dmso-d6)δ7.27(d,j＝7.6hz,1h),6.06(s,1h),6.01(s,1h),5.72(s,1h),4.98(s,1h),4.13(d,j＝4.4hz,1h),3.91(ddd,j＝9.2,7.5,5.7hz,1h),3.80
–
3.75(m,1h),3.38(dd,j＝7.1,2.2hz,1h),3.13
–
3.08(m,1h),2.83(dd,j＝18.7,5.2hz,1h),2.47(d,j＝3.4hz,1h),1.99(s,1h),1.94(d,j＝13.6hz,1h),1.66(dt,j＝8.9,5.1hz,1h),1.57(dd,j＝8.0,6.3hz,1h),1.54(d,j＝3.4hz,1h),1.51
–
1.47(m,2h),1.47
–
1.44(m,1h),1.41(s,1h),1.39(s,2h),1.35(s,9h),1.27
–
1.22(m,1h),1.14(s,1h),1.12(s,2h),1.10(s,1h),0.88(s,3h),0.84(s,1h),0.82(d,j＝4.7hz,7h),0.79(s,1h).
13
c nmr(101mhz,dmso-d6)δ202.86,200.85,172.00,155.28,146.32,121.99,97.39,78.12,76.07,64.86,61.86,57.38,55.66,52.19,47.28,46.57,42.80,39.82,38.58,33.86,31.90,
29.91,28.07,27.73,24.22,23.22,22.48,21.46,20.53,18.06,14.69,14.31.
[0040]
实施例10
[0041][0042]
用95mg 2-甲基烟酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为62％。1h nmr(400mhz,dmso-d6)δ8.64(dd,j＝4.9,1.8hz,1h),8.05(dd,j＝7.9,1.8hz,1h),7.38(dd,j＝7.9,4.8hz,1h),6.34(d,j＝1.0hz,1h),6.16(d,j＝1.4hz,1h),5.79(s,1h),5.04(s,1h),4.20
–
4.15(m,1h),3.86(qd,j＝7.1,3.1hz,1h),3.47
–
3.41(m,1h),3.41
–
3.38(m,1h),2.95(dd,j＝18.7,5.2hz,1h),2.66(s,3h),2.57(dd,j＝18.5,2.4hz,1h),2.48(d,j＝12.3hz,1h),2.10(s,1h),2.05
–
1.97(m,1h),1.69(dt,j＝14.3,5.2hz,1h),1.59
–
1.51(m,1h),1.48(dd,j＝11.8,5.3hz,1h),1.43
–
1.38(m,3h),1.20(t,j＝7.0hz,3h),1.12
–
1.07(m,1h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.76,201.07,164.88,158.59,152.21,146.31,137.95,124.33,122.57,121.51,97.59,77.03,65.25,62.11,57.26,55.91,47.21,46.52,42.90,39.78,38.75,33.86,31.87,29.93,24.49,23.26,20.49,18.08,14.77.
[0043]
实施例11
[0044][0045]
用98mg 5-氟烟酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为80％。1h nmr(400mhz,dmso-d6)δ8.88(d,j＝2.8hz,1h),8.85(d,j＝1.6hz,1h),8.07(ddd,j＝8.8,2.9,1.7hz,1h),6.38(d,j＝1.0hz,1h),6.17(d,j＝1.4hz,1h),5.79(s,1h),5.03(s,1h),4.28
–
4.24(m,1h),3.86(dq,j＝9.4,7.0hz,1h),3.45
–
3.40(m,1h),3.40
–
3.37(m,1h),2.93(dd,j＝18.7,5.1hz,1h),2.60
–
2.54(m,1h),2.51(p,j＝1.8hz,1h),2.11(s,1h),2.03
–
1.97(m,1h),1.68(dt,j＝14.1,5.1hz,1h),1.55(ddd,j＝14.1,10.1,7.5hz,1h),1.48(dd,j＝11.7,5.3hz,1h),1.42(s,2h),1.40
–
1.36(m,1h),1.19(t,j＝7.0hz,3h),1.13(td,j＝5.3,2.4hz,1h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.64,200.82,162.61,159.96,157.41,146.05,142.76,126.54,123.37,122.69,97.57,77.43,65.09,62.06,57.34,55.95,47.21,46.52,42.89,39.76,38.69,33.85,31.86,29.91,23.25,20.45,18.07,14.73.
[0046]
实施例12
[0047][0048]
用95mg 5-甲基烟酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为28％。1h nmr(400mhz,dmso-d6)δ8.79(d,j＝2.0hz,1h),8.68(d,j＝2.1hz,1h),8.02(t,j＝2.4hz,1h),6.35(s,1h),6.17(d,j＝1.4hz,1h),5.78(s,1h),5.02(s,1h),4.22(d,j＝4.5hz,1h),3.88
–
3.81(m,1h),3.46
–
3.39(m,1h),3.38
–
3.35(m,1h),2.93(dd,j＝18.7,5.2hz,1h),2.60
–
2.54(m,1h),2.49(d,j＝8.1hz,1h),2.37(s,3h),2.11(s,1h),2.04
–
1.98(m,1h),1.69(dt,j＝14.1,5.1hz,1h),1.60
–
1.53(m,1h),1.48(dd,j＝11.7,5.2hz,1h),1.42(s,2h),1.40(s,1h),1.19(t,j＝7.0hz,3h),1.12(dd,j＝16.4,6.2hz,1h),0.87(s,3h),0.82(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.76,200.99,163.81,154.41,147.15,146.15,136.74,133.56,124.56,122.63,97.57,76.92,65.17,62.05,57.31,55.92,47.20,46.56,42.89,39.76,38.72,33.86,31.88,29.92,23.29,20.47,18.07,17.60,14.74.
[0049]
实施例13
[0050][0051]
用42mg乙酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为64％。1h nmr(400mhz,dmso-d6)δ6.10(d,j＝1.4hz,1h),6.03(s,1h),5.75(s,1h),4.99(s,1h),4.10
–
4.06(m,1h),3.79(dq,j＝9.3,7.0hz,1h),3.38(dq,j＝9.3,7.0hz,1h),3.16(ddt,j＝4.1,2.7,1.3hz,1h),2.83(dd,j＝18.6,5.1hz,1h),2.47
–
2.42(m,1h),2.01(s,3h),1.98(d,j＝11.9hz,2h),1.66(dt,j＝14.1,5.1hz,1h),1.52(ddd,j＝13.7,10.1,7.3hz,1h),1.47
–
1.43(m,1h),1.42(s,1h),1.39(s,2h),1.35
–
1.21(m,1h),1.13(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.89,201.16,169.41,146.40,122.29,97.44,75.54,64.99,61.93,57.26,55.61,47.21,46.57,42.83,39.80,38.66,33.87,31.88,29.93,23.20,20.64,20.49,18.07,14.67.
[0052]
实施例14
[0053][0054]
用86mg 4-嘧啶甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为76％。1h nmr(400mhz,dmso-d6)δ9.40(d,j＝1.4hz,1h),9.10(d,j＝5.1hz,1h),7.98(dd,j＝5.1,1.4hz,1h),6.40
–
6.39(m,1h),6.16(d,j＝1.4hz,1h),5.79(s,1h),5.02(s,
1h),4.20
–
4.18(m,1h),3.85(dq,j＝9.5,7.1hz,1h),3.45
–
3.38(m,2h),2.95(dd,j＝18.7,5.2hz,1h),2.60(d,j＝2.4hz,1h),2.56
–
2.53(m,1h),2.12(s,1h),2.04
–
1.98(m,1h),1.68(dt,j＝14.2,5.1hz,1h),1.56(dt,j＝9.0,7.2hz,1h),1.49(dd,j＝11.7,5.3hz,1h),1.43(d,j＝6.5hz,2h),1.40(s,1h),1.19(t,j＝7.0hz,3h),1.12
–
1.07(m,1h),0.87(s,3h),0.82(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.89,201.16,158.09,153.21,149.11,145.63,140.68,122.70,113.27,97.57,77.44,65.12,62.08,57.36,55.96,47.16,46.64,42.90,39.77,38.65,33.87,31.88,29.92,23.17,20.47,18.06,14.76.
[0055]
实施例15
[0056][0057]
用86mg 2-吡嗪甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为51％。1h nmr(400mhz,dmso-d6)δ9.14(d,j＝1.4hz,1h),8.92(d,j＝2.4hz,1h),8.85
–
8.81(m,1h),6.41(s,1h),6.17
–
6.14(m,1h),5.79(s,1h),5.02(s,1h),4.21(d,j＝4.4hz,1h),3.86(dq,j＝9.5,7.1hz,1h),3.45
–
3.39(m,2h),2.95(dd,j＝18.7,5.2hz,1h),2.58(dd,j＝18.7,2.4hz,1h),2.47(s,1h),2.11(s,1h),2.01(d,j＝14.1hz,1h),1.68(dt,j＝14.1,5.1hz,1h),1.61
–
1.53(m,1h),1.49(dd,j＝11.7,5.3hz,1h),1.43
–
1.40(m,2h),1.39(s,1h),1.20(t,j＝7.0hz,3h),1.09(t,j＝7.2hz,1h),0.87(s,3h),0.81(d,j＝4.4hz,3h).
13
cnmr(101mhz,dmso-d6)δ202.73,200.89,162.67,148.43,146.12,145.58,144.92,142.43,122.70,97.57,77.44,65.12,62.08,57.36,55.96,47.16,46.64,42.90,39.77,38.65,33.87,31.88,29.92,23.17,20.47,18.06,14.76.
[0058]
实施例16
[0059][0060]
用78mg糠酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为78％。1h nmr(400mhz,dmso-d6)δ7.99(d,j＝1.8hz,1h),7.28(dd,j＝3.6,0.8hz,1h),6.69(dd,j＝3.5,1.7hz,1h),6.25(d,j＝1.0hz,1h),6.14(d,j＝1.4hz,1h),5.78(s,1h),5.01(s,1h),4.14
–
4.11(m,1h),3.81(dq,j＝9.3,7.0hz,1h),3.40(dq,j＝9.4,7.0hz,1h),2.91(dd,j＝18.7,5.2hz,1h),2.56(d,j＝2.4hz,1h),2.48(d,j＝14.8hz,1h),2.09(s,1h),2.00(dq,j＝14.5,2.8hz,1h),1.68(dt,j＝14.1,5.1hz,1h),1.59
–
1.51(m,1h),1.47(dd,j＝11.7,5.4hz,1h),1.41(d,j＝7.2hz,2h),1.40
–
1.37(m,1h),1.27
–
1.19(m,1h),1.16(t,j＝7.0hz,3h),1.11(d,j＝20.1hz,1h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.78,200.96,156.77,148.10,146.16,143.04,122.58,119.22,112.46,97.54,76.45,65.16,62.04,57.29,55.84,47.16,46.59,42.88,39.76,38.70,33.87,31.89,29.91,
23.20,20.47,18.05,14.73.
[0061]
实施例17
[0062][0063]
用112mg 3-吲哚甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为25％。1h nmr(400z,chloroform-d)δ8.52(dd,j＝7.5,1.4hz,1h),8.06(s,2h),7.61(td,j＝7.4,1.6hz,1h),7.47(td,j＝7.5,1.7hz,1h),7.34(dd,j＝7.4,1.5hz,1h),6.49(dd,j＝13.8,1.0hz,1h),6.23(dd,j＝13.8,1.0hz,1h),6.03(d,j＝7.1hz,1h),5.35(s,1h),4.25(t,j＝7.1hz,1h),3.41
–
3.33(m,2h),3.11(dq,j＝12.1,6.0hz,1h),2.67(dd,j＝14.0,7.1hz,1h),2.54(s,1h),2.16(dd,j＝14.0,7.1hz,1h),2.09(dt,j＝13.0,7.1hz,1h),1.94
–
1.89(m,1h),1.87(dt,j＝13.0,4.6hz,1h),1.74
–
1.63(m,2h),1.57
–
1.49(m,1h),1.47
–
1.40(m,1h),1.28(dt,j＝12.9,7.0hz,1h),1.22(t,j＝7.0hz,1h),1.15(t,j＝6.0hz,3h),0.95(s,3h),0.89(s,3h).
13
c nmr(101hz,chloroform-d)δ208.26,205.26,164.40,152.28,136.63,132.80,127.71,122.44,121.78,121.00,119.55,112.27,102.69,100.30,74.02,68.96,64.96,60.40,49.56,45.30,42.87,42.26,41.37,41.29,36.27,33.63,32.25,25.19,21.50,19.67,15.25.
[0064]
实施例18
[0065][0066]
用124mg 4-甲氧基肉桂酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为68％。1h nmr(400mhz,dmso-d6)δ7.70(d,j＝1.7hz,1h),7.68(d,j＝2.3hz,1h),7.61(d,j＝15.9hz,1h),6.98(s,1h),6.95(s,1h),6.47(d,j＝16.0hz,1h),6.14(s,1h),6.14
–
6.11(m,1h),5.77(s,1h),5.01(s,1h),4.16(d,j＝4.4hz,1h),3.87
–
3.81(m,1h),3.80(s,3h),3.39(dq,j＝9.3,7.0hz,1h),3.28
–
3.23(m,1h),2.88(dd,j＝18.7,5.2hz,1h),2.54(t,j＝3.1hz,1h),2.01(d,j＝17.5hz,2h),1.68(dt,j＝14.0,5.1hz,1h),1.58
–
1.49(m,1h),1.48
–
1.44(m,1h),1.43(d,j＝6.1hz,1h),1.40(s,2h),1.35
–
1.22(m,1h),1.16(t,j＝7.0hz,3h),1.10(d,j＝8.3hz,1h),0.87(s,3h),0.82(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.98,201.28,165.61,161.27,146.48,145.28,130.36(*2),126.43,122.37,114.56,114.33(*2),97.48,75.69,65.08,61.92,57.33,55.75,55.31,47.22,46.62,42.87,39.79,38.79,33.88,31.90,29.95,26.31,23.23,20.50,18.07,14.73.
[0067]
实施例19
[0068][0069]
用134mg 4-甲基肉桂酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为72％。1h nmr(400mhz,dmso-d6)δ7.64
–
7.59(m,3h),7.24(s,1h),7.22(s,1h),6.56(d,j＝16.1hz,1h),6.15(s,1h),6.13(d,j＝1.4hz,1h),5.77(s,1h),5.01(s,1h),4.17(d,j＝4.4hz,1h),3.81(dq,j＝9.3,7.0hz,1h),3.40(dt,j＝9.3,7.0hz,1h),3.25(d,j＝4.7hz,1h),2.88(dd,j＝18.6,5.2hz,1h),2.54(dd,j＝4.8,3.1hz,1h),2.33(s,3h),2.04(s,1h),1.99(s,1h),1.68(dt,j＝14.0,5.1hz,1h),1.58
–
1.50(m,1h),1.49
–
1.45(m,1h),1.42(s,1h),1.39(d,j＝2.3hz,2h),1.24(d,j＝3.5hz,1h),1.16(t,j＝7.0hz,4h),0.87(s,3h),0.82(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.97,201.25,165.45,146.44,145.46,140.70,131.09,129.50(*2),128.52(*2),122.41,116.24,97.49,75.82,65.07,61.93,57.32,55.75,47.20,46.62,42.87,38.77,33.88,31.90,29.95,23.22,21.00,20.50,18.07,14.73.
[0070]
实施例20
[0071][0072]
用103mg肉桂酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为80％。1h nmr(400mhz,dmso-d6)δ7.74(d,j＝2.0hz,1h),7.74
–
7.72(m,1h),7.66(d,j＝16.0hz,1h),7.43(d,j＝1.9hz,2h),7.42(d,j＝2.8hz,1h),6.63(d,j＝16.0hz,1h),6.18(s,1h),6.14(d,j＝1.4hz,1h),5.78(s,1h),5.01(s,1h),4.18(d,j＝4.4hz,1h),3.83(dq,j＝9.5,7.0hz,1h),3.40(dq,j＝9.4,7.0hz,1h),3.29
–
3.25(m,1h),2.89(dd,j＝18.7,5.2hz,1h),2.57
–
2.53(m,1h),2.04(s,1h),2.03
–
1.97(m,1h),1.68(dt,j＝14.1,5.1hz,1h),1.58
–
1.51(m,1h),1.49
–
1.44(m,1h),1.42(s,1h),1.39(d,j＝2.8hz,2h),1.17(t,j＝7.0hz,4h),1.12(d,j＝11.5hz,1h),0.88(s,3h),0.82(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.90,201.21,165.32,146.44,145.46,133.79,130.63,128.86,128.50,122.40,117.38,97.50,75.92,65.09,61.97,57.35,55.77,47.24,46.63,42.88,39.79,38.78,33.87,31.90,29.95,23.23,20.49,18.08,14.73.
[0073]
实施例21
[0074]
[0075]
用123mg 3-硝基肉桂酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为56％。1h nmr(400mhz,dmso-d6)δ8.61(t,j＝1.9hz,1h),8.27
–
8.21(m,2h),7.81(d,j＝16.1hz,1h),7.70(t,j＝8.0hz,1h),6.89(d,j＝16.1hz,1h),6.19(s,1h),6.15(d,j＝1.4hz,1h),5.79(s,1h),5.02(s,1h),4.22(d,j＝4.4hz,1h),3.83(dq,j＝9.3,7.0hz,1h),3.41(dq,j＝9.3,7.0hz,1h),3.31
–
3.27(m,1h),2.91(dd,j＝18.6,5.2hz,1h),2.56(t,j＝3.5hz,1h),2.05
–
1.98(m,2h),1.69(dt,j＝14.0,5.1hz,1h),1.54(ddd,j＝13.9,10.1,7.4hz,1h),1.49
–
1.44(m,1h),1.44
–
1.42(m,1h),1.41(s,2h),1.18(t,j＝7.0hz,4h),1.12(dt,j＝8.9,5.2hz,1h),0.88(s,3h),0.82(s,3h).13c nmr(101mhz,dmso-d6)δ202.85,201.12,164.97,148.25,146.37,142.98,135.72,134.34,130.27,124.71,123.14,122.43,120.47,97.51,76.19,65.05,61.97,57.37,55.76,47.24,46.62,42.87,39.78,38.74,33.86,31.89,29.94,23.22,20.47,18.07,14.72.
[0076]
实施例22
[0077][0078]
用109mg 4-吡啶丙烯酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为76％。1h nmr(400mhz,dmso-d6)δ8.64
–
8.61(m,2h),7.72
–
7.69(m,2h),7.64(d,j＝16.1hz,1h),6.90(d,j＝16.1hz,1h),6.20(s,1h),6.15
–
6.13(m,1h),5.78(s,1h),5.01(s,1h),4.20
–
4.16(m,1h),3.85
–
3.76(m,1h),3.40(dd,j＝7.9,6.2,5.5hz,1h),3.30
–
3.27(m,1h),2.89(dd,j＝18.7,5.2hz,1h),2.56
–
2.52(m,1h),2.07(s,1h),2.01(d,j＝13.9hz,1h),1.68(m,j＝14.1,5.1hz,1h),1.59
–
1.51(m,1h),1.50
–
1.45(m,1h),1.41(t,j＝6.0hz,3h),1.25(d,j＝10.2hz,1h),1.18(s,1h),1.17(s,2h),1.15(s,1h),0.88(s,3h),0.82(s,3h).13c nmr(101mhz,dmso)δ202.86,201.13,164.80,150.31(*2),146.32,142.84,140.91,122.54,122.24(*2),122.08,97.52,76.32,65.05,61.98,57.35,55.76,47.19,46.64,42.88,41.23,38.72,33.88,31.90,29.93,23.21,20.50,18.06,14.73.
[0079]
实施例23
[0080][0081]
用110mg 2-甲基-5-嘧啶甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为70％。1h nmr(400mhz,chloroform-d)δ9.08(s,2h),6.53(d,j＝1.4hz,1h),6.31
–
6.26(m,1h),5.62(d,j＝1.2hz,1h),5.07(s,1h),4.23(dd,j＝4.9,1.1hz,1h),3.98(q,j＝9.4,7.0hz,1h),3.46(q,j＝9.3,7.0hz,1h),3.32(dd,j＝5.4,2.7,1.3hz,1h),3.07(dt,j＝18.5,5.0,1.0hz,1h),2.67
–
2.58(m,2h),2.12
–
2.04(m,1h),1.95(s,1h),1.80(dt,j＝14.4,5.1hz,1h),1.64(td,j＝14.0,4.2hz,1h),1.58
–
1.53(m,1h),1.44(d,j＝5.4hz,1h),1.41(d,j＝5.1hz,1h),1.39
–
1.32(m,1h),1.28(t,j＝7.1hz,3h),0.92(s,3h),
0.87(s,3h).
13
c nmr(101mhz,cdcl3)δ202.91,200.97,172.40,162.93,158.06,146.07,122.62,120.75,98.43,77.61,77.22,66.20,63.34,58.67,56.74,48.83,46.84,43.72,40.29,39.51,34.31,32.24,30.30,23.73,20.72,18.59,14.95.
[0082]
实施例24
[0083][0084]
用141mg 5-溴-2-嘧啶甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为62％。1h nmr(400mhz,chloroform-d)δ9.08(s,2h),6.53(d,j＝1.4hz,1h),6.31
–
6.26(m,1h),5.62(d,j＝1.2hz,1h),5.07(s,1h),4.23(dd,j＝4.9,1.1hz,1h),3.98(q,j＝9.4,7.0hz,1h),3.46(q,j＝9.3,7.0hz,1h),3.32(dd,j＝5.4,2.7,1.3hz,1h),3.07(dt,j＝18.5,5.0,1.0hz,1h),2.67
–
2.58(m,2h),2.12
–
2.04(m,1h),1.95(s,1h),1.80(dt,j＝14.4,5.1hz,1h),1.64(td,j＝14.0,4.2hz,1h),1.58
–
1.53(m,1h),1.44(d,j＝5.4hz,1h),1.41(d,j＝5.1hz,1h),1.39
–
1.32(m,1h),1.28(t,j＝7.1hz,3h),0.92(s,3h),0.87(s,3h).
13
c nmr(101mhz,cdcl3)δ202.91,200.97,172.40,162.93,158.06,146.07,122.62,120.75,98.43,77.61,77.22,66.20,63.34,58.67,56.74,48.83,46.84,43.72,40.29,39.51,34.31,32.24,30.30,23.73,20.72,18.59,14.95
[0085]
实施例25
[0086][0087]
用104mg反-3-吡啶丙烯酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为73％。1h nmr(400mhz,dmso-d6)δ8.89(d,j＝2.2hz,1h),8.59(dd,j＝4.8,1.6hz,1h),8.21(dt,j＝8.1,2.0hz,1h),7.71(d,j＝16.1hz,1h),7.44(dd,j＝8.0,4.8hz,1h),6.80(d,j＝16.1hz,1h),6.19(s,1h),6.14(s,1h),5.79(s,1h),5.01(s,1h),4.18(d,j＝4.4hz,1h),3.82(dq,j＝9.3,7.1hz,1h),3.44
–
3.37(m,1h),3.28(d,j＝4.7hz,1h),2.90(dd,j＝18.7,5.2hz,1h),2.57
–
2.53(m,1h),2.06(s,1h),2.04
–
1.99(m,1h),1.68(dt,j＝14.1,5.1hz,1h),1.58
–
1.50(m,1h),1.49
–
1.45(m,1h),1.43
–
1.38(m,3h),1.28(d,j＝16.3hz,1h),1.17(t,j＝7.0hz,4h),0.88(s,3h),0.82(s,3h).
13
c nmr(101mhz,dmso)δ203.40,201.68,165.53,151.60,150.56,146.86,142.75,135.31,130.17,124.36,123.01,119.88,97.99,76.60,65.55,62.45,57.83,56.24,47.68,47.13,43.37,40.27,39.24,34.37,32.39,30.43,23.70,20.98,18.56,15.22.
[0088]
实施例26
[0089][0090]
用104mg反-2-吡啶丙烯酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为59％。1h nmr(400mhz,chloroform-d)δ8.72(d,j＝2.2hz,1h),8.61(dd,j＝4.9,1.6hz,1h),7.82(dt,j＝8.0,2.0hz,1h),7.64(d,j＝16.1hz,1h),7.33(dd,j＝8.0,4.8hz,1h),6.46(d,j＝16.1hz,1h),6.35(s,1h),6.27(d,j＝1.4hz,1h),5.61(d,j＝1.2hz,1h),5.06(s,1h),4.27(dd,j＝4.9,1.1hz,1h),4.01
–
3.94(m,1h),3.45(dq,j＝9.2,7.0hz,1h),3.28(dh,j＝3.9,1.3hz,1h),3.05(ddd,j＝18.5,5.0,1.0hz,1h),2.62(ddd,j＝7.2,2.6,1.1hz,1h),2.60
–
2.54(m,1h),2.09(dq,j＝14.4,2.8hz,1h),1.92(s,1h),1.80(dt,j＝14.4,5.1hz,1h),1.76
–
1.73(m,1h),1.63(td,j＝13.9,4.1hz,1h),1.57
–
1.52(m,1h),1.41(ddd,j＝10.9,7.1,4.2hz,2h),1.28(d,j＝7.0hz,3h),1.25(d,j＝2.2hz,1h),0.93(s,3h),0.87(s,3h).
13
c nmr(101mhz,cdcl3)δ203.40,201.63,165.25,151.24,149.80,146.45,141.99,134.25,129.88,123.77,122.33,119.57,98.32,76.64,66.09,63.19,58.64,56.62,48.84,46.95,43.70,40.33,39.42,34.33,32.27,30.36,23.76,20.75,18.61,14.94.
[0091]
实施例27
[0092][0093]
用59mg丙酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为63％。1h nmr(400mhz,dmso-d6)δ6.10(d,j＝1.4hz,1h),6.05(d,j＝1.1hz,1h),5.74(t,j＝0.9hz,1h),4.99(s,1h),4.08
–
4.05(m,1h),3.80(dq,j＝9.3,7.0hz,1h),3.42
–
3.35(m,1h),3.32(s,1h),3.15(dt,j＝3.9,2.6,1.2hz,1h),2.83(dd,j＝18.8,4.9hz,1h),2.46(dt,j＝2.5,1.1hz,1h),2.36
–
2.25(m,2h),2.00(d,j＝3.9hz,1h),1.99
–
1.93(m,1h),1.66(dt,j＝14.1,5.1hz,1h),1.52(dd,j＝13.7,10.1,7.2hz,1h),1.45(d,j＝5.4hz,1h),1.42(s,1h),1.39(s,2h),1.13(t,j＝7.0hz,4h),0.99(t,j＝7.5hz,3h),0.87(s,3h),0.81(s,3h).
13
cnmr(101mhz,dmso)δ202.92,201.15,172.66,146.41,122.23,97.44,75.36,65.02,61.93,57.24,55.67,47.20,46.54,42.83,39.79,38.70,33.88,31.88,29.93,26.74,23.21,20.50,18.07,14.69,8.85.
[0094]
实施例28
[0095]
[0096]
用63mg丙烯酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为56％。1h nmr(400mhz,dmso-d6)δ6.30(dd,j＝17.2,1.5hz,1h),6.19
–
6.13(m,1h),6.11(t,j＝1.5hz,2h),5.97(dd,j＝10.3,1.6hz,1h),5.97(dd,j＝1hz,1h),5.00(s,1h),4.09
–
4.06(m,1h),3.79(dq,j＝9.3,7.0hz,1h),3.39(dq,j＝9.3,7.0hz,1h),3.32(s,1h),3.22(ddd,j＝5.3,2.5,1.3hz,1h),2.91
–
2.82(m,1h),2.54
–
2.52(m,1h),2.48
–
2.43(m,1h),2.04(s,1h),2.02
–
1.96(m,1h),1.67(dt,j＝14.1,5.1hz,1h),1.57
–
1.49(m,1h),1.49
–
1.44(m,1h),1.41(dt,j＝11.5,3.5hz,3h),1.14(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ202.83,201.06,164.43,146.26,132.43,127.73,122.44,97.49,75.93,65.06,61.97,57.23,55.73,47.17,46.54,42.85,39.77,38.67,33.87,31.88,29.92,23.19,20.49,18.05,14.70.
[0097]
实施例29
[0098][0099]
用62mg丁酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为58％。1h nmr(400mhz,dmso-d6)δ6.10(d,j＝1.4hz,1h),6.05
–
6.01(m,1h),5.75(s,1h),4.99(s,1h),4.08
–
4.04(m,1h),3.78(dq,j＝9.3,7.0hz,1h),3.38(dq,j＝9.3,7.0hz,1h),3.16(ddd,j＝5.4,2.5,1.3hz,1h),2.84(dd,j＝18.7,5.2hz,1h),2.48
–
2.42(m,1h),2.34
–
2.20(m,2h),1.99(d,j＝11.4hz,2h),1.66(dt,j＝14.1,5.1hz,1h),1.50(q,j＝7.3hz,3h),1.47
–
1.44(m,1h),1.40(d,j＝10.8hz,3h),1.13(t,j＝7.0hz,4h),0.87(d,j＝1.6hz,4h),0.85(s,2h),0.83(s,1h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ202.87,201.15,171.82,146.42,122.18,97.45,75.43,65.08,61.91,57.24,55.63,47.23,46.52,42.82,39.80,38.72,35.32,33.86,31.87,29.93,23.22,20.51,18.06,17.98,14.68,13.14.
[0100]
实施例30
[0101][0102]
用81mg 2-乙基丁酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为48％。1h nmr(400mhz,dmso-d6)δ6.09(d,j＝1.3hz,1h),6.00(d,j＝1.0hz,1h),5.75(s,1h),5.00(s,1h),4.06
–
4.04(m,1h),3.78
–
3.71(m,1h),3.41
–
3.36(m,1h),3.16(dt,j＝5.3,1.8hz,1h),2.85(dd,j＝18.7,5.2hz,1h),2.47(d,j＝2.3hz,1h),2.20
–
2.13(m,1h),2.03(s,1h),2.02
–
1.96(m,1h),1.67(dt,j＝14.1,5.1hz,1h),1.47(dq,j＝6.4,2.4,1.8hz,4h),1.42(dd,j＝7.3,2.9hz,4h),1.39(s,2h),1.30
–
1.22(m,1h),1.13(d,j＝7.0hz,4h),0.87(s,3h),0.81(d,j＝1.6hz,3h),0.79
–
0.76(m,3h).
13
c nmr(101mhz,dmso)δ203.39,201.67,174.75,146.95,122.56,97.97,76.06,65.72,62.33,57.69,56.05,48.75,
47.73,46.93,43.30,40.30,39.29,34.36,32.38,30.43,25.20,25.11,23.77,21.08,18.55,15.21,11.86,11.82.
[0103]
实施例31
[0104][0105]
用69mg环丙乙酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为64％。1h nmr(400mhz,dmso-d6)δ6.00(d,j＝1.3hz,1h),5.95(d,j＝1.1hz,1h),5.65(d,j＝1.0hz,1h),4.90(s,1h),4.00
–
3.97(m,1h),3.71(dq,j＝9.2,7.0hz,1h),3.33
–
3.26(m,1h),3.07(ddt,j＝4.0,2.7,1.3hz,1h),2.78
–
2.70(m,1h),2.38
–
2.35(m,1h),2.20
–
2.04(m,2h),1.89(d,j＝13.4hz,2h),1.56(dt,j＝14.0,5.1hz,1h),1.42(ddd,j＝13.7,10.1,7.2hz,1h),1.38
–
1.34(m,1h),1.32(s,1h),1.31
–
1.27(m,2h),1.24
–
1.13(m,1h),1.04(t,j＝7.0hz,4h),0.84
–
0.79(m,1h),0.78(s,3h),0.71(s,3h),0.33(ddd,j＝8.1,4.6,1.5hz,2h),-0.00(tt,j＝5.7,2.5hz,2h).
13
c nmr(101mhz,dmso)δ203.45,201.62,171.91,146.93,122.66,97.93,75.92,65.54,62.40,57.69,56.16,47.68,47.00,43.32,40.29,39.24,38.72,34.37,32.37,30.43,23.72,21.02,18.56,15.21,7.20,4.55,4.47.
[0106]
实施例32
[0107][0108]
用151mg boc-5氨基戊酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为79％。1h nmr(400mhz,dmso-d6)δ6.11
–
6.09(m,1h),6.03(s,1h),5.74(t,j＝1.0hz,1h),4.99(s,1h),4.07
–
4.04(m,1h),3.78(dq,j＝9.3,7.0hz,1h),3.42
–
3.35(m,1h),3.18
–
3.12(m,1h),2.91
–
2.86(m,2h),2.83(dd,j＝17.2,3.6hz,1h),2.46(t,j＝1.7hz,1h),2.35
–
2.23(m,2h),1.98(d,j＝14.1hz,2h),1.66(dt,j＝14.0,5.1hz,1h),1.56
–
1.49(m,1h),1.46(t,j＝5.8hz,2h),1.43(d,j＝2.4hz,1h),1.42(s,2h),1.39(s,1h),1.37(s,9h),1.33(d,j＝7.1hz,2h),1.28(d,j＝15.6hz,1h),1.24(d,j＝3.4hz,1h),1.13(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ203.44,201.64,172.41,156.02,146.88,122.75,97.92,77.80,75.91,65.54,62.38,57.72,56.12,47.67,47.01,43.31,40.27,39.68,39.20,34.37,33.53,32.37,30.43,29.13,28.72(*3),23.72,22.20,21.01,18.55,15.19.
[0109]
实施例33
[0110][0111]
用141mg boc-γ氨基丁酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为67％。1h nmr(400mhz,dmso-d6)δ6.83(t,j＝5.7hz,1h),6.09(d,j＝1.4hz,1h),6.05(s,1h),5.74(s,1h),4.99(s,1h),4.07(d,j＝4.4hz,1h),3.80(dq,j＝9.2,7.0hz,1h),3.38(dt,j＝9.4,7.0hz,1h),3.18
–
3.14(m,1h),2.90(td,j＝6.4,5.9,2.1hz,2h),2.83(dd,j＝18.7,5.1hz,1h),2.49
–
2.41(m,2h),2.29(td,j＝15.8,15.1,8.4hz,2h),1.99(s,2h),1.66(dt,j＝14.0,5.1hz,1h),1.57(t,j＝7.2hz,2h),1.46(t,j＝6.2hz,1h),1.42(d,j＝5.3hz,2h),1.39(s,1h),1.37(s,9h),1.28(d,j＝15.7hz,1h),1.23(s,1h),1.13(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ203.49,201.67,172.18,156.04,146.87,122.79,97.90,77.94,75.90,65.50,62.37,57.71,56.15,47.64,47.04,43.32,40.27,39.40,39.16,34.38,32.38,31.26,30.43,28.69(*3),25.29,23.70,21.01,18.56,15.21.
[0112]
实施例34
[0113][0114]
用141mg boc-2-氨基丁酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为71％。1h nmr(400mhz,dmso-d6)δ7.25(d,j＝7.4hz,1h),6.06(s,1h),6.01(s,1h),5.72(s,1h),4.99(s,1h),4.13(d,j＝4.4hz,1h),3.83(d,j＝7.2hz,1h),3.80
–
3.73(m,2h),3.42
–
3.34(m,2h),3.11(d,j＝4.9hz,1h),2.83(dd,j＝18.7,5.3hz,1h),2.46(s,1h),1.99(s,2h),1.98
–
1.94(m,1h),1.65(dt,j＝13.8,5.2hz,2h),1.57(d,j＝7.4hz,2h),1.50(dd,j＝13.5,7.6hz,2h),1.45(d,j＝5.2hz,1h),1.42(d,j＝4.9hz,3h),1.35(s,9h),1.30
–
1.22(m,3h),1.12(d,j＝2.2hz,3h),0.88(s,3h),0.83(s,2h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ203.49,201.67,172.18,156.04,146.87,122.79,97.90,79.94,75.90,65.50,62.37,57.71,56.15,47.64,46.54,43.32,40.27,39.40,39.16,32.38,31.26,30.43,28.69(*3),24.29,23.70,21.01,18.56,15.21,10.93.
[0115]
实施例35
[0116][0117]
用161mg boc-氨基己酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产
物，产率为84％。1h nmr(400mhz,dmso-d6)δ6.73(t,j＝5.7hz,1h),6.10(d,j＝1.4hz,1h),6.02(d,j＝1.1hz,1h),5.75(d,j＝1.1hz,1h),4.99(s,1h),4.08
–
4.04(m,1h),3.78(dq,j＝9.2,7.0hz,1h),3.42
–
3.35(m,1h),3.17
–
3.13(m,1h),2.88(d,j＝6.7hz,1h),2.86
–
2.79(m,2h),2.46(t,j＝1.7hz,1h),2.28(q,j＝7.4hz,2h),1.98(d,j＝16.3hz,2h),1.67(dt,j＝14.1,5.1hz,1h),1.56
–
1.50(m,1h),1.50
–
1.46(m,2h),1.45(s,1h),1.42(d,j＝5.6hz,2h),1.39(s,1h),1.37(s,9h),1.33(s,1h),1.30(d,j＝4.8hz,1h),1.25
–
1.17(m,3h),1.13(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ203.45,201.66,172.43,155.98,146.89,122.75,97.94,77.77,75.93,65.56,62.40,57.69,56.12,47.67,47.00,43.32,40.27,40.09,39.19,34.37,33.91,32.37,30.42,29.51,28.73(*3),25.99,24.64,23.70,21.01,18.56,15.20.
[0118]
实施例36
[0119][0120]
用149mg n-boc-吡咯烷酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为87％。1h nmr(400mhz,dmso-d6)δ6.11(d,j＝1.4hz,1h),6.06(s,1h),5.75(s,1h),4.99(s,1h),4.03(d,j＝4.4hz,1h),3.83
–
3.72(m,1h),3.42
–
3.35(m,3h),3.21(d,j＝7.1hz,2h),3.18(d,j＝4.7hz,2h),2.83(dd,j＝18.7,5.2hz,1h),2.46(dd,j＝5.8,2.0hz,1h),2.03(s,1h),1.98(d,j＝13.9hz,2h),1.91(q,j＝6.5hz,1h),1.66(dt,j＝14.0,4.9hz,1h),1.46(q,j＝6.6,5.7hz,1h),1.42(s,1h),1.41(s,1h),1.39(d,j＝2.1hz,9h),1.25(d,j＝9.6hz,1h),1.12(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).δ
13
c nmr(101mhz,dmso)δ203.35,201.47,172.48,153.65,146.77,122.79,97.98,78.90,76.52,65.59,65.55,62.45,57.70,56.18,47.64,46.95,43.31,42.93,42.07,40.60,40.18,39.12,34.36,32.37,30.40,28.90(*3),23.73,20.99,18.55,15.21.
[0121]
实施例37
[0122][0123]
用139mg n-boc-环丁烷甲酸代替4-氯甲基苯甲酸，其他操作同实施例2，得白色固体产物，产率为61％。1h nmr(400mhz,dmso-d6)δ6.12(d,j＝1.0hz,1h),6.11
–
6.09(m,1h),5.75(t,j＝1.0hz,1h),5.00(s,1h),4.06
–
4.01(m,2h),3.98(d,j＝12.5hz,1h),3.85
–
3.76(m,3h),3.45(dt,j＝8.8,5.5hz,1h),3.40(ddd,j＝9.4,6.3,2.8hz,1h),3.26
–
3.22(m,1h),2.84(dd,j＝18.7,5.1hz,1h),2.53(d,j＝2.4hz,1h),2.49
–
2.47(m,1h),2.43(d,j＝12.6hz,1h),2.03(s,1h),1.98(d,j＝13.8hz,1h),1.66(dt,j＝14.0,5.1hz,1h),1.53(dd,j＝10.8,6.9hz,1h),1.46(q,j＝6.2hz,1h),1.42(s,2h),1.37(s,9h),1.22(d,j＝9.7hz,
1h),1.14(t,j＝7.0hz,4h),0.87(s,3h),0.80(s,3h).
13
c nmr(101mhz,dmso)δ203.36,201.49,171.94,155.81,146.72,122.95,97.98,79.44,76.75,65.49,62.46,57.78,56.19,47.60,47.02(*2),43.33,40.59,40.38,40.17,39.04,34.38,32.37,31.99,30.41,28.45(*3),23.67,21.01,18.55,15.22.
[0124]
实施例38
[0125][0126]
将所得11-羰基-20-乙氧基济源冬凌草甲素14-位l-boc-苯丙氨酸酯化衍生物溶于二氯甲烷中，加入过量的三氟乙酸使其脱掉boc保护基并成盐，将体系旋蒸浓缩后加入异丙醚使其析出得到目标衍生物。1h nmr(400mhz,dmso-d6)δ8.78(s,3h),7.37(d,j＝7.7hz,1h),7.26(d,j＝1.5hz,1h),7.22(s,2h),7.21
–
7.20(m,1h),6.06(s,1h),6.03(s,1h),5.69(s,1h),5.01(s,1h),4.23(d,j＝4.4hz,1h),4.13(q,j＝7.6hz,1h),3.83(dq,j＝9.3,7.0hz,1h),3.44
–
3.37(m,1h),3.06
–
3.01(m,1h),2.89(d,j＝7.8hz,2h),2.86
–
2.79(m,1h),2.46(d,j＝2.3hz,1h),1.98(d,j＝9.0hz,2h),1.67(dt,j＝13.8,4.8hz,1h),1.57
–
1.49(m,1h),1.42(s,2h),1.42
–
1.39(m,2h),1.19(t,j＝7.2hz,1h),1.15(s,1h),1.13(s,2h),1.11(s,1h),0.89(d,j＝3.9hz,3h),0.83(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.96,201.56,171.00,146.12,137.21,129.02,128.11(*2),126.41,122.12,97.40,76.22,64.80,61.88,57.46,55.67,55.56,47.31,46.59,42.82,41.23,39.82,38.50,36.33,33.85,31.90,29.93,23.23,20.70,18.08,14.69.
[0127]
实施例39
[0128][0129]
用boc-丙氨酸代替l-boc-苯丙氨酸，其他操作同实施例37，得白色固体产物，产率为71％。1h nmr(400mhz,dmso-d6)δ8.35(s,3h),6.06(s,1h),6.01(s,1h),5.72(s,1h),4.99(s,1h),4.13(d,j＝4.5hz,1h),3.97(t,j＝7.2hz,1h),3.82
–
3.76(m,1h),3.37(dd,j＝9.5,7.2hz,1h),3.10(d,j＝4.7hz,1h),2.83(dd,j＝18.6,5.1hz,1h),2.45(d,j＝13.2hz,1h),1.99(s,1h),1.65(dt,j＝14.1,5.0hz,1h),1.55
–
1.48(m,1h),1.45(d,j＝5.3hz,1h),1.42(d,j＝4.8hz,2h),1.39(s,2h),1.23(dd,j＝8.6,6.6hz,1h),1.17(d,j＝7.3hz,3h),1.14(s,1h),1.12(s,2h),1.11(s,1h),0.88(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso-d6)δ202.89,200.87,172.15,146.26,122.08,97.39,76.03,64.82,61.88,57.36,55.66,49.13,47.24,46.56,42.81,39.80,38.57,33.86,31.89,29.91,23.20,20.51,18.06,16.67,14.70.
[0130]
实施例40
[0131][0132]
用boc-氨基戊酸代替l-boc-苯丙氨酸，其他操作同实施例37，得白色固体产物，产率为71％。1h nmr(400mhz,dmso-d6)δ7.80(s,3h),6.11
–
6.09(m,1h),6.03(s,1h),5.74(t,j＝1.0hz,1h),4.99(s,1h),4.07
–
4.04(m,1h),3.78(dq,j＝9.3,7.0hz,1h),3.42
–
3.35(m,1h),3.18
–
3.12(m,1h),2.91
–
2.86(m,2h),2.85(dd,j＝17.2,3.6hz,1h),2.46(t,j＝1.7hz,1h),2.35
–
2.23(m,2h),1.98(d,j＝14.1hz,2h),1.66(dt,j＝14.0,5.1hz,1h),1.56
–
1.49(m,1h),1.46(t,j＝5.8hz,2h),1.43(d,j＝2.4hz,1h),1.42(s,2h),1.39(s,1h),1.33(d,j＝7.1hz,2h),1.28(d,j＝15.6hz,1h),1.24(d,j＝3.4hz,1h),1.13(t,j＝7.0hz,3h),0.86(s,3h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ203.44,201.64,172.41,146.88,122.75,97.92,75.91,65.54,62.38,57.72,56.12,47.67,47.01,43.31,40.27,39.68,39.20,34.37,33.53,32.37,30.43,29.13,23.72,22.20,21.01,18.55,15.19.
[0132]
实施例41
[0133][0134]
用boc-2-氨基丁酸代替l-boc-苯丙氨酸，其他操作同实施例37，得白色固体产物，产率为71％。1h nmr(400mhz,dmso-d6)δ8.46(s,3h),6.07(s,1h),6.01(s,1h),5.73(s,1h),4.99(s,1h),4.13(d,j＝4.4hz,1h),3.82(d,j＝7.2hz,1h),3.80
–
3.73(m,2h),3.42
–
3.34(m,2h),3.11(d,j＝4.9hz,1h),2.83(dd,j＝18.7,5.3hz,1h),2.46(s,1h),1.99(s,2h),1.98
–
1.94(m,1h),1.65(dt,j＝13.8,5.2hz,2h),1.57(d,j＝7.4hz,2h),1.50(dd,j＝13.5,7.6hz,2h),1.45(d,j＝5.2hz,1h),1.42(d,j＝4.9hz,3h),1.29
–
1.22(m,3h),0.88(s,3h),0.83(s,2h),0.81(s,3h).
13
c nmr(101mhz,dmso)δ203.49,201.67,172.18,146.87,122.79,97.90,75.90,65.50,62.37,57.71,56.15,47.64,46.54,43.32,40.27,39.40,39.16,32.38,31.26,30.43,24.29,23.70,21.01,18.56,15.21,10.93.
[0135]
实施例42
[0136][0137]
用n-boc-环丁烷甲酸代替l-boc-苯丙氨酸，其他操作同实施例37，得白色固体产物，产率为61％。1h nmr(400mhz,dmso-d6)δ9.86(s,1h),8.89(s,1h),6.12(d,j＝1.0hz,1h),6.11
–
6.09(m,1h),5.75(t,j＝1.0hz,1h),5.00(s,1h),4.06
–
4.01(m,2h),3.98(d,j＝12.5hz,1h),3.85
–
3.76(m,3h),3.45(dt,j＝8.8,5.5hz,1h),3.40(ddd,j＝9.4,6.3,2.8hz,1h),3.26
–
3.22(m,1h),2.84(dd,j＝18.7,5.1hz,1h),2.53(d,j＝2.4hz,1h),2.49
–
2.47(m,1h),2.43(d,j＝12.6hz,1h),2.03(s,1h),1.98(d,j＝13.8hz,1h),1.66(dt,j＝14.0,5.1hz,1h),1.53(dd,j＝10.8,6.9hz,1h),1.46(q,j＝6.2hz,1h),1.42(s,2h),1.22(d,j＝9.7hz,1h),1.14(t,j＝7.0hz,4h),0.87(s,3h),0.80(s,3h).
13
c nmr(101mhz,dmso)δ203.36,201.49,171.94,146.72,122.95,97.98,76.75,65.49,62.46,57.78,56.19,47.60,47.02(*2),43.33,40.59,40.38,40.17,39.04,34.38,32.37,31.99,30.41,23.67,21.01,18.55,15.22.
[0138]
实施例43
[0139][0140]
用n-boc-吡咯烷酸代替l-boc-苯丙氨酸，其他操作同实施例37，得白色固体产物，产率为87％。1h nmr(400mhz,dmso-d6)δ9.58(s,1h),8.42(s,1h),6.11(d,j＝1.4hz,1h),6.06(s,1h),5.75(s,1h),4.99(s,1h),4.03(d,j＝4.4hz,1h),3.83
–
3.72(m,1h),3.42
–
3.35(m,3h),3.21(d,j＝7.1hz,2h),3.18(d,j＝4.7hz,2h),2.83(dd,j＝18.7,5.2hz,1h),2.46(dd,j＝5.8,2.0hz,1h),2.03(s,1h),1.98(d,j＝13.9hz,2h),1.91(q,j＝6.5hz,1h),1.66(dt,j＝14.0,4.9hz,1h),1.46(q,j＝6.6,5.7hz,1h),1.42(s,1h),1.41(s,1h),1.25(d,j＝9.6hz,1h),1.12(t,j＝7.0hz,4h),0.87(s,3h),0.81(s,3h).δ
13
c nmr(101mhz,dmso)δ203.35,201.47,172.48,146.77,122.79,97.98,76.52,65.59,65.55,62.45,57.70,56.18,47.64,46.95,43.31,42.93,42.07,40.60,40.18,39.12,34.36,32.37,30.40,23.73,20.99,18.55,15.21.
[0141]
实验方法：以下百分含量若有特别说明均为质量百分含量，以11-羰基-20-乙氧基济源冬凌草甲素为阳性对照，选用以上五种癌细胞测定所合成的化合物72小时抗增殖活性：将对数生长期的细胞用含0.25％edta的胰酶消化之后配制成浓度为1-10
×
104个/ml的细胞悬液，按照1000-10000个细胞/孔接种于96孔板中，每孔快速顶壁加入100μl细胞悬液，接种完毕后，将96孔板放置在体积百分含量5％co2的37℃湿度培养箱中。24h后弃干净板中上清液，每孔加入含有不同药物浓度的培养液200μl，每个化合物按照一定浓度梯度设置三个平行孔，对照组加空白的培养液200μl，再放入体积百分含量5％co2的37℃湿度培养箱中培养72h。每孔加入100μl 30％tca溶液于4℃下固定60min；用去离子水洗3-4次，风干；每孔加入0.4％srb 100μl室温染色30min；用1％的乙酸洗3次，风干；每孔加入150μl的tris base后用平板摇床摇15min,待摇匀后，使用酶标仪测定吸光密度值(od),检测波长540nm。以溶剂对照处理细胞为对照组，按下述公式计算化合物对细胞的抑制率，并按中效方程计算半数抑制浓度ic
50
：抑制率(％)＝(对照组od均值-给药组od均值)/对照组od均值*100％
[0142]
11-羰基-20-乙氧基济源冬凌草甲素酯化衍生物抗肿瘤活性
ori为冬凌草甲素；joa为济源冬凌草甲素；joa11为11-羰基-20-乙氧基济源冬凌草甲素。上述实验结果表明：本发明所述化合物具有较好的体外抗肿瘤活性，可用于制备抗肿瘤药物，应用于临床治疗食管癌、胃癌、宫颈癌、胰腺癌等。以本发明化合物作为活性成分用于制备新的抗癌药物，具有潜在的应用价值。
[0143]
药物溶解度试验根据《中国药典》2020年版溶解度试验，本发明选用生理盐水、甲醇、乙醇、丙酮、乙酸乙酯、二氯甲烷和石油醚为溶剂，结果表明大部分化合物在二氯甲烷中易溶，其溶解度可达到200mg/ml。
[0144]
药物稳定性试验根据《中国药典》2020年版药物稳定性试验指导原则，对抗肿瘤活性最好的化合物10进行稳定性试验，结果如下表所示：化合物10影响因素试验(1批供试品)
化合物10加速试验和长期试验(3批供试品)