具体实施方式
为了理解本发明,下面以实施例进一步说明本发明,但不意于限制本发明的保护范围。
实施例1:化合物1-50的制备方法
化合物1的制备:
将小白菊内酯(50mg,0.2mmol)溶于2.5mL二氯甲烷中,加入对甲苯磺酸(5mg,0.026mmol)。反应体系放置于室温,搅拌过夜。将反应液转到NaHCO3(10mL)饱和溶液中,收集有机相,水相用少量的二氯甲烷萃取,将有机相合到一起,用Na2SO4干燥,过滤,再用旋转蒸发仪减压蒸馏有机溶剂,硅胶柱纯化,得到化合物1(45mg,收率90%)。1H NMR(CDCl3,400MHz)δ6.20(d,J=3.2Hz,1H)5.49(d,J=3.2Hz,1H)3.81(t,J=10.4Hz,1H),2.70(d,J=10.4Hz,1H),2.65-2.62(m,2H),2.40-2.34(m,1H),2.07-2.26(m,4H),1.73-1.86(m,2H),1.68(s,3H),1.36-1.28(m,4H);13C NMR(CDCl3,100MHz)δ169.8,138.7,131.7,130.8,119.5,84.1,80.2,58.5,49.5,38.2,34.8,30.0,25.7,23.9,23.6。
化合物2-20的合成通法:
氮气保护下,冰水浴下,将酰氯(RCl)滴加到化合物1(24.8mg,0.1mmol)和DMAP(1.25mg,0.01mmol)和三乙胺(0.12mL,1.2mmol)的混合液中,滴加结束后撤去冰浴,常温搅拌,直至TLC检测反应物反应完毕。将反应混合物倾入到冰水中,用乙酸乙酯(5mL×3)萃取,有机层依次用柠檬酸溶液(20mL),饱和NaHCO3(10mL),饱和食盐水(10mL)洗涤,有机相用无水硫酸钠干燥,减压浓缩得粗品,硅胶柱层析分离,得到纯品。
化合物2,产率为62.8%。1H NMR(400MHz,CDCl3)δ6.13(1H,d,J=3.1Hz),5.40(1H,d,J=2.8Hz),3.72(1H,t,J=10.2Hz),3.08(1H,d,J=10.1Hz),2.65~2.59(1H,m),2.41~2.35(2H,m),2.21~2.13(4H,m),2.04~2.00(1H,m),1.97(3H,s),1.91~1.83(1H,m),1.65(3H,s),1.48(3H,s);13C NMR(100MHz,CDCl3)δ169.6,169.3,138.4,130.6,129.4,117.8,87.7,82.0,55.6,49.1,35.4,33.9,29.4,24.9,23.2,21.5,17.8。
化合物3,产率为70.0%,1H NMR(400MHz,CDCl3)δ6.20(1H,d,J=3.1Hz),5.47(1H,d,J=2.8Hz),3.79(1H,t,J=0.1Hz),3.13(1H,d,J=10.2Hz),2.71~2.66(1H,m),2.49~2.42(2H,m),2.38~2.30(2H,m),2.28~2.26(4H,m),2.11~2.08(1H,dd,J=13.8,1.8Hz),1.97~1.89(1H,m),1.72(3H,s),1.57(6H,d,J=9.0Hz);13C NMR(100MHz,CDCl3)δ173.8,170.2,139.5,131.5,130.4,118.6,88.4,83.0,56.6,50.1,36.5,34.9,30.4,28.7,25.9,24.1,18.8,9.1。
化合物4,产率为30.0%,1H NMR(400MHz,CDCl3)δ6.22(1H,d,J=2.7Hz),5.49(1H,d,J=2.1Hz),3.81(1H,t,J=10.1Hz),3.16(1H,d,J=10.3Hz),2.52~2.44(2H,m),2.36~2.23(6H,m),2.13~2.10(1H,m),1.99~1.91(1H,m),1.74(3H,s),1.70~1.68(1H,m),1.65(3H,s),1.57(2H,s),0.98(3H,t,J=7.3Hz);13C NMR(100MHz,CDCl3)δ173.2,170.3,139.5,131.6,130.5,118.7,88.5,83.0,56.8,50.2,37.4,36.6,35.0,30.5,25.9,24.2,18.8,18.5,13.6。
化合物5,产率为37.6%,1H NMR(400MHz,CDCl3)δ6.19(1H,d,J=3.1Hz),5.47(1H,d,J=2.9Hz),3.78(1H,t,J=10.1Hz),3.13(1H,d,J=10.0Hz),2.72~2.66(1H,m),2.49~2.42(2H,m),2.37~2.31(1H,m),2.30~2.26(4H,m),2.12~2.08(1H,dd,J=13.7,2.2Hz),1.97~1.88(1H,m),1.75(3H,s),1.64~1.56(2H,m),1.55(3H,s),1.40~1.31(3H,m),0.92(3H,t,J=7.3Hz);13C NMR(100MHz,CDCl3)δ173.4,170.2,139.5,131.6,130.4,118.7,88.5,83.0,56.8,50.2,36.5,35.2,34.9,30.5,27.1,25.9,24.1,22.2,18.8,13.8。
化合物6,产率为26.8%,1H NMR(400MHz,CDCl3)δ6.21(1H,d,J=3.0Hz),5.48(1H,d,J=2.8Hz),3.81(1H,t,J=10.1Hz),3.15(1H,d,J=10.3Hz),2.74~2.68(1H,m),2.57~2.44(3H,m),2.30~2.29(3H,m),2.14~2.10(1H,dd,J=13.7,2.1Hz),1.98~1.90(1H,m),1.74(3H,s),1.66(1H,s),1.56(3H,s),1.21~1.17(6H,m);13C NMR(100MHz,CDCl3)δ176.6,170.2,139.6,131.5,130.4,118.6,88.2,83.0,56.8,50.1,36.5,35.0,34.7,30.5,26.0,24.2,19.0,18.9,18.7。
化合物7,产率为47.8%,1H NMR(400MHz,CDCl3)δ6.19(1H,d,J=3.2Hz),5.46(1H,d,J=2.9Hz),3.78(1H,t,J=10.1Hz),3.13(1H,d,J=10.1Hz),2.71~2.66(1H,m),2.50~2.42(2H,m),2.28~2.26(3H,m),2.20~2.07(4H,m),1.97~1.89(1H,m),1.72~1.69(4H,m),1.56(3H,s),0.97(6H,d,J=5.9Hz);13C NMR(100MHz,CDCl3)δ172.7,170.2,139.5,131.6,130.5,118.7,88.5,83.0,56.8,50.2,44.6,36.6,35.0,30.5,25.9,25.8,24.2,22.4,22.3,18.8。
化合物8,产率为13.9%,1H NMR(400MHz,CDCl3)δ6.20(1H,d,J=2.9Hz),5.47(1H,d,J=2.4Hz),4.07~3.95(2H,q,J=16.4Hz),3.79(1H,t,J=10.1Hz),3.47(3H,s),3.19(1H,d,J=10.1Hz),2.71~2.66(1H,m),2.51~2.44(2H,m),2.27(3H,s),2.11~2.08(1H,dd,J=12.5,0.7Hz),2.04~1.95(1H,m),1.72(3H,s)1.63(1H,s),1.58(3H,s);13C NMR(100MHz,CDCl3)δ170.2,169.7,139.3,131.9,130.0,118.9,89.6,82.9,70.3,59.3,56.4,50.0,36.4,35.0,30.4,25.9,24.2,18.9。
化合物9,产率为56.2%,1H NMR(400MHz,CDCl3)δ6.23(1H,d,J=3.3Hz),5.50(1H,d,J=3.1Hz),4.14~4.05(2H,m),3.81(1H,t,J=10.1Hz),3.20(1H,d,J=10.0Hz),2.74~2.68(1H,m),2.54~2.47(2H,m),2.30~2.29(3H,m),2.15~2.11(1H,dd,J=13.8,2.3Hz),2.07~1.97(1H,m),1.75(3H,s),1.62(4H,s);13C NMR(100MHz,CDCl3)δ170.1,166.4,139.2,132.1,129.8,119.0,90.8,82.8,56.4,50.1,42.0,36.3,35.0,30.3,25.9,24.2,18.8。
化合物10,产率为20.7%,1H NMR(400MHz,CDCl3)δ6.23(1H,d,J=3.3Hz),5.96(1H,s),5.51(1H,d,J=3.0Hz),3.81(1H,t,J=10.1Hz),3.21(1H,d,J=10.1Hz),2.76~2.70(1H,m),2.56~2.48(2H,m),2.30(3H,s),2.15~2.11(1H,dd,J=13.8,2.3Hz),2.07~1.99(1H,m),1.75(3H,s),1.66(4H,s);13C NMR(100MHz,CDCl3)δ169.9,163.4,139.2,132.4,129.5,119.0,92.3,82.5,65.2,56.5,50.1,36.0,34.9,30.2,25.9,24.1,18.6。
化合物11,产率为46.5%,1H NMR(400MHz,CDCl3)δ6.22(1H,d,J=3.3Hz),5.49(1H,d,J=3.0Hz),3.86(2H,s),3.80(1H,t,J=10.1Hz),3.18(1H,d,J=9.9Hz),2.74~2.68(1H,m),2.53~2.46(2H,m),2.30~2.29(3H,m),2.14~2.10(1H,dd,J=13.8,2.3Hz),2.06~1.94(1H,m),1.74(3H,s),1.71(1H,s),1.60(3H,s);13C NMR(100MHz,CDCl3)δ170.1,166.3,139.3,132.1,129.9,118.9,90.8,82.8,56.5,50.1,36.2,34.9,30.3,27.7,25.9,24.2,18.7。
化合物12,产率为25.8%,1H NMR(400MHz,CDCl3)δ6.22(1H,d,J=3.2Hz),5.49(1H,d,J=2.8Hz),3.81(1H,t,J=10.2Hz),3.47(2H,t,J=6.5Hz),3.15(1H,d,J=10.0Hz),2.73~2.68(1H,m),2.51~2.44(2H,m),2.40~2.32(2H,m),2.30(2H,d,J=6.4Hz),2.14~2.10(1H,dd,J=13.7,1.9Hz),1.99~1.91(3H,m),1.83~1.78(2H,m),1.74(3H,s),1.60(3H,s),1.57(2H,s);13C NMR(100MHz,CDCl3)δ172.5,170.2,139.4,131.7,130.2,118.8,88.75,83.0,56.7,50.1,36.5,35.0,34.5,33.5,32.0,30.5,25.9,24.2,23.6,18.8。
化合物13,产率为47.6%,1H NMR(400MHz,CDCl3)δ6.22(1H,d,J=3.3Hz),5.49(1H,d,J=3.0Hz),3.81(1H,t,J=10.2Hz),3.37~3.33(2H,m),3.16(1H,d,J=10.1Hz),2.74~2.68(1H,m),2.51~2.45(2H,m),2.41~2.33(2H,m),2.31~2.29(3H,m),2.14~2.10(1H,dd,J=13.7,2.2Hz),2.00~1.92(1H,m),1.74(3H,s),1.73~1.66(4H,m),1.62(1H,s),1.57(3H,s);13C NMR(100MHz,CDCl3)δ171.5,169.2,138.4,130.7,129.2,117,7,87.7,82.0,55.7,50.1,49.1,35.5,34.0,33.9,29.4,27.2,24.9,23.1,21.2,17.8。
化合物14,产率为78.2%,1H NMR(400MHz,CDCl3)δ6.47~6.37(1H,dd,J=17.3,1.3Hz),6.21(1H,d,J=3.3Hz),6.13~6.06(1H,m),5.80~5.77(1H,dd,J=10.3,1.3Hz),5.48(1H,d,J=3.1Hz),3.82(1H,t,J=10.1Hz),3.15(1H,d,J=10.1Hz),2.73~2.67(1H,m),2.58~2.44(2H,m),2.29~2.27(3H,m),2.12~2.08(1H,dd,J=13.7,2.3Hz),2.00~1.92(1H,m),1.73(3H,s),1.59(4H,s);13C NMR(100MHz,CDCl3)δ170.3,165.5,139.4,131.7,130.2,130.1,130.0,118.8,88.8,83.0,57.1,50.1,36.5,35.0,30.5,25.9,24.2,18.6。
化合物15,产率为13.5%,1H NMR(400MHz,CDCl3)δ6.22(1H,d,J=3.3Hz),5.49(1H,d,J=3.0Hz),5.20(1H,d,J=5.0Hz),5.16(1H,s),3.81(1H,t,J=10.1Hz),3.16(1H,d,J=8.8Hz),3.12~3.09(2H,m),2.74~2.68(1H,m),2.51~2.44(2H,m),2.30~2.28(4H,m),2.14~2.10(1H,dd,J=13.7,2.1Hz),2.00~1.92(1H,m),1.88~1.86(1H,dd,J=6.9,1.3Hz),1.74(3H,s),1.59(1H,s),1.58(3H,s);13C NMR(100MHz,CDCl3)δ171.0,170.2,139.5,131.7,130.7,130.3,118.7,118.2,89.0,83.0,56.7,50.1,40.2,36.5,35.0,30.5,25.9,24.2,18.8。
化合物16,产率为26.6%,1H NMR(400MHz,CDCl3)δ6.20(1H,d,J=3.2Hz),5.68(1H,s),5.47(1H,d,J=2.8Hz),3.81(1H,t,J=10.2Hz),3.15(1H,d,J=10.0Hz),2.73~2.67(1H,m),2.57~2.43(2H,m),2.28~2.27(3H,m),2.15(3H,s),2.12~2.08(1H,dd,J=13.9,2.1Hz),2.00~1.92(2H,m),1.87(3H,s),1.73(3H,s),1.59(3H,s);13C NMR(100MHz,CDCl3)δ170.3,166.2,155.4,139.5,131.5,130.6,118.7,117.7,88.2,83.1,57.0,50.3,36.8,34.9,30.6,27.4,25.9,24.2,20.1,18.9。
化合物17,产率为23.3%,1H NMR(400MHz,CDCl3)δ7.76(1H,d,J=16.0Hz),7.59~7.57(2H,m),7.39(3H,d,J=5.0Hz),6.45(1H,d,J=16.0Hz),6.24(1H,d,J=3.1Hz),5.51(1H,d,J=2.7Hz),3.88(1H,t,J=10.2Hz),3.20(1H,d,J=10.0Hz),2.78~2.73(1H,m),2.65~2.61(1H,m),2.55~2.48(1H,m),2.31(3H,s),2.16(1H,d,J=13.5Hz),2.07~1.98(1H,q),1.75(3H,s),1.63(4H,s);13C NMR(100MHz,CDCl3)δ170.4,166.4,144.5,139.5,134.7,131.6,130.0,129.9,128.8,128.2,119.7,118.8,88.7,83.1,57.3,50.0,36.7,35.0,30.6,26.0,24.2,18.6。
化合物18,产率为53.3%,1H NMR(400MHz,CDCl3)δ6.18(1H,d,J=3.2Hz),5.87~5.80(1H,m),5.46(1H,d,J=3.0Hz),5.09(1H,d,J=16.9Hz),5.00(1H,d,J=9.6Hz),3.80~3.74(1H,m),3.12(1H,d,J=7.4Hz),2.68(1H,s),2.45~2.40(3H,m),2.37(4H,s),2.25(3H,s),2.11~2.07(1H,m),1,95~1.89(1H,m),1.71(3H,s),1.54(3H,d,J=3.8Hz);13C NMR(100MHz,CDCl3)δ172.4,170.2,139.5,136.9,131.6,130.3,118.7,115.3,88.7,82.9,56.7,50.1,36.5,35.0,34.6,30.4,29.0,25.9,24.2,18.8。
化合物19,产率为65.2%,1H NMR(400MHz,CDCl3)δ6.21(1H,d,J=3.1Hz),5.49(1H,d,J=2.7Hz),3.82(1H,t,J=10.0Hz),3.15(1H,d,J=9.3Hz),2.91~2.82(3H,m),2.14~2.10(1H,m),2.04~2.01(3H,m),1.97~1.87(8H,m),1.83~1.75(6H,m),1.63(1H,s);13C NMR(100MHz,CDCl3)δ172.4,170.2,139.4,131.7,130.2,118.8,88.8,83.6,83.0,69.0,56.7,50.1,36.5,34.9,34.1,30.5,25.9,24.2,23.7,18.8,17.8。
化合物20,产率为24.0%,1H NMR(400MHz,CDCl3)δ7.32~7.18(5H,m),6.22(1H,d,J=3.3Hz),5.49(1H,d,J=3.0Hz),3.80(1H,t,J=10.1Hz),3.12(1H,d,J=10.1Hz),2.99(2H,t,J=7.9Hz),2.73~2.59(3H,m),2.49~2.43(2H,m),2.30~2.28(3H,m),2.14~2.10(1H,dd,J=13.8,2.3Hz),1.96~1.87(1H,m),1.74(3H,s),1.65(1H,s),1.56(3H,s);13C NMR(100MHz,CDCl3)δ172.3,170.2,140.8,139.5,131.6,130.4,128.4,126.0,118.7,88.8,83.0,56.7,50.1,36.8,36.5,35.0,31.0,30.5,26.0,24.2,18.8。
化合物21的合成
在一干燥洁净的10mL反应瓶内,加入5-乙炔酸与化合物19(200mg,0.584mmol)、6-叠氮-1-正己醇(125.43mg,0.876mmol)、水合硫酸铜(145.80mg,0.584mmol)和抗坏血酸钠(462.78mg,2.336mmol),加入蒸馏水与叔丁醇(1∶2)混合溶液(3mL)溶解成均相,常温搅拌2小时。用旋转蒸发仪将部分溶剂旋出,剩余反应混合溶液用乙酸乙酯萃取,收集有机相并用无水硫酸钠干燥,减压抽滤。用旋转蒸发仪将有机相旋干,硅胶柱层析色谱分离,得到化合物21(120.0mg),产率为61.3%。1H NMR(400MHz,CDCl3)δ7.52(1H,s),6.20(1H,d,J=3.3Hz),5.49(1H,d,J=3.0Hz),4.34(2H,t,J=7.1Hz),3.81(1H,t,J=10.2Hz),3.63(2H,t,J=6.4Hz),3.14(1H,d,J=10.1Hz),2.81~2.76(2H,m),2.74~2.67(1H,m),2.51~2.43(2H,m),2.39~2.31(2H,m),2.29~2.27(4H,m),2.13~2.09(1H,dd,J=13.8,2.3Hz),2.05~2.00(3H,m),1.97~1.88(5H,m),1.72(4H,s),1.60~1.53(6H,m);13CNMR(100MHz,CDCl3)δ172.6,170.2,147.1,139.4,131.7,130.0,121.3,118.8,88.6,83.1,62.2,56.7,50.0,49.9,36.4,34.9,34.7,32.3,30.4,30.1,26.1,25.8,25.1,24.8,24.7,24.1,18.8。
化合物22的制备:
将二甲基胺盐酸盐(1.5g,18mmol)与K2CO3(5.0g,36mmol)混合加入100ml二氯甲烷中搅拌15min,然后加压过滤,直接加入到化合物5(300mg,1.2mmol)中,在室温下搅拌3h。减压除去溶剂,再用少量二氯溶解,用水快速洗三遍,Na2SO4干燥,过滤,减压除去二氯甲烷,得到的粗产物二甲基胺中间体再次溶于少量二氯甲烷中,边搅拌边加入稀盐酸溶液(与二甲基胺中间体等当量),搅拌期间要测量水溶液的pH值,当水溶液pH值为4-5时,停止滴加盐酸水溶液。收集水相,冻干,得到化合物22。
二甲基胺中间体:1H NMR(CDCl3,400MHz)δ3.76(t,J=10.0Hz,1H),2.96(s,1H),2.49-2.67(m,3H),2.28-2.34(m,1H),2.30-2.34(m,2H),2.18(s,6H),2.09(br s,2H),1.96(d,J=11.2Hz,1H)1.67-1.73(m,2H),1.60(s,3H),1.22(br s,3H),1.18(brs,2H);13C NMR(CDCl3,300MHz)δ177.0,131.8,131.3,84.0,80.2,58.3,58.1,50.9,46.0,44.6,38.4,35.3,30.0,27.2,23.7,22.8.
化合物22:[α]D 20=-42.0(c=10,H2O);IR(KBr):3334,2927,2856,1767,1467,992,967,874,831,719,669,626,504cm-1;1H NMR (D2O,400MHz)δ4.14(t,J=10.3Hz,1H),3.51(q,J=12.6Hz,1H),3.40(dd,J=13.3,2.9Hz,1H),3.18-3.04(m,1H),2.96(d,J=10.6Hz,6H),2.67(d,J=10.2Hz,1H),2.37(dd,J=16.2,8.1Hz,1H),2.27-2.05(m,4H),1.87(d,J=12.9Hz,1H),1.73(dd,J=19.5,11.7Hz,2H),1.66(s,3H),1.46-1.31(m,2H),1.26(s,3H);13C NMR(CDCl3,100MHz)δ178.4,132.6,131.4,85.1,80.7,56.9,55.6,49.9,45.1,42.3,41.5,39.2,34.4,29.5,25.9,23.2,21.4.HRMS calcd for C17H27NO3[M+H]+294.1991,found 294.2069.
化合物23的制备:
将二甲基胺盐酸盐(1.5g,18mmol)与K2CO3(5.0g,36mmol)混合加入100ml二氯甲烷中搅拌15min,然后加压过滤,直接加入到化合物5(300mg,1.2mmol)中,在室温下搅拌3h。减压除去溶剂,再用少量二氯溶解,用水快速洗三遍,Na2SO4干燥,过滤,减压除去二氯甲烷,得到的粗产物二甲基胺中间体再次溶于少量二氯甲烷中,边搅拌边加入富马酸(与二甲基胺中间体等当量),浓缩干燥得到化合物23。1H NMR(DMSO,400MHz)δ6.58(s,2H),3.80(t,J=10.3Hz,1H),2.64(s,3H),2.49-2.53(m,3H),2.26-2.27(m,1H),2.23(s,6H),1.96-2.10(m,6H),1.60(s,3H),1.57-1.59(m,2H),1.23-1.25(m,1H),1.15(s,3H);13C NMR(CDCl3,100MHz)δ177.8,167.6,135.2,133.7,131.4,83.4,80.31,58.0,57.1,51.7,45.2,43.6,41.0,35.3,30.2,27.0,24.2,23.2.
化合物24的制备:
将化合物1(628mg,12.5mmol)溶于35ml二氯甲烷中,然后加入m-CPBA(680mg,4.0mmol)。反应体系放置于室温搅拌,并用TCL监测反应。原料消失以后,将反应混合液倾倒在5%NaHCO3(60mL)中,有机相再用水洗(20ml),收集有机相,并用Na2SO4干燥,过滤,减压旋蒸,得到粗产品用硅胶柱纯化得到化合物24。1H NMR(CDCl3,400MHz)δ6.17(d,J=3.2Hz,1H),5.47(d,J=2.8Hz,1H),4.04(t,J=10.8Hz,1H),2.36-2.20(m,4H),2.03-1.08(m,4H),1.68-1.62(m,1H),1.46(s,3H),1.46(d,J=12.8Hz,1H),1.29(s,3H);13C NMR(CDCl3,100MHz)δ169.5,137.9,119.5,81.7,79.5,69.7,62.1,55.3,49.2,37.2,33.2,29.3,23.1,23.0,21.8。
化合物25的制备:
根据文献尹宏权,齐秀兰,华会明,裴月湖,云木香化学成分研究,中国药物化学杂志,2005年15卷4期,217-220。从云木香中分离纯化得到。
化合物26和30的制备:
化合物25(150mg,0.65mmol),甲醇钠(20mg,0.37mmol)的甲醇溶液在30℃下搅拌10小时。TLC检测完成后,将反应液倒入冰水中,用乙酸乙酯萃取四次,合并乙酸乙酯层,依次用5%盐酸,饱和碳酸氢钠水溶液,饱和食盐水洗涤,无水硫酸镁干燥,过滤后浓缩过柱纯化得到甲氧基化产物(136mg,80%)。
1H-NMR(400MHz,CDCl3)δ5.19(d,J=1.6Hz,1H),5.03(d,J=1.6Hz,1H),4.87(s,1H),4.76(s,1H),3.93(t,J=9.2Hz,1H),3.70(dd,J=4.4,9.8Hz,1H),3.63(dd,J=3.2,9.8Hz,1H),3.37(s,3H),2.90(m,1H),2.83(m,1H),2.51(m,2H),2.49(m,1H),2.44(m,1H),2.38(m,1H),2.19(m,1H),2.07(m,1H),1.95(m,1H),1.86(m,1H),1.32(m,1H);13C-NMR(100MHz,CDCl3)δ175.7,151.8,149.9,111.5,108.8,85.3,68.9,59.0,51.7,47.7,46.9,43.9,37.7,32.5,32.4,30.1.
在连冷凝管及干燥管的两口瓶中加入Zn-Cu齐(812mg)及无水乙醚(4mL),加入一小粒碘,搅拌直至碘的颜色消失。加入甲氧基化产物(262mg,1mmol)和二碘甲烷(0.8mL.10mmol)的乙醚(2mL)溶液,回流72小时。TLC显示反应完全后,倾出乙醚溶液,残留的固体用乙醚洗涤(3mL)两次,合并乙醚层,分别用饱和氯化铵水溶液,水(10mL)洗涤两次,用无水硫酸钠干燥,过滤浓缩,过柱纯化得到产物为一白色固体化合物30(206mg,71%)。1H-NMR(400MHz,CDCl3)δ4.20(m,1H),3.65(dd,J=3.9,9.9Hz,1H),3.59(dd,J=3.0,9.9Hz,1H),3.34(s,1H),2.30-2.40(m,3H),1.92-2.07(m,2H),1.15-1.70(m,7H),0.85(m,1H),0.70(m,1H),0.50(m,1H),0.40(m,1H),0.16-0.36(m,4H);13C-NMR(100MHz,CDCl3)δ176.4,83.9,69.2,59.2,51.5,47.9,47.3,44.3,36.2,35.9,29.0,27.3,26.7,18.5,12.5,10.4,10.0,8.8;ESI-HRMS m/z:291.1964[M+H]。
化合物30(0.023g,0.080mmol)溶于乙腈(0.52mL),加入4M NaOH水溶液(0.11mL),回流5小时,TLC检测完全,冷却至室温,慢慢加入10%盐酸至pH为3。加入乙酸乙酯(20mL),用水洗涤两次(2×20mL),水相再用乙酸乙酯萃取两次(2×20mL),合并有机层,无水硫酸钠干燥,过滤,浓缩过柱得到产物为一白色固体26(0.019g,94%)。
分子式:C17H22O2
分子量:258
性状:白色无定型粉末
谱图数据:
1H-NMR(400MHz,CDCl3)δ6.17(d,J=3.2Hz,1H),5.45(d,J=3.2Hz,1H),4.24(dd,J=8.8,10.8Hz,1H),2.87(m,1H),2.20(m,1H),2.07(m,1H),1.95(dd,J=8.8,10.4Hz,1H),1.71(m,2H),1.37-1.61(m,5H),0.98(m,1H),0.64(m,1H),0.49(m,1H),0.43(m,1H),0.27-0.37(m,4H);13C NMR(100MHz,CDCl3)δ169.4,139.4,118.5,82.5,51.6,47.4,44.2,34.2,32.8,26.4,26.2,25.3,17.4,11.1,10.7,10.5,7.8;ESI-HRMS m/z:259.1692[M+H]。
化合物27的制备
二甲胺盐酸盐(245mg)悬浮于11mL二氯甲烷溶液中,加入碳酸钾(380mg)搅拌半小时,加入化合物26(42mg),回流4小时,硅胶柱层析(石油醚∶乙酸乙酯:=70∶30),得油状化合物27,收率:78%。
分子式:C19H29NO2
分子量:303
性状:油状液体
谱图数据:
1H-NMR(400MHz,CDCl3)δ4.19(dd,J=9.5,10.2Hz,1H),2.66(dd,J=12.8,4.8Hz,1H),2.49(dd,J=12.8,6.8Hz,1H),2.25-2.38(m,2H),2.21(s,6H),1.98-2.12(m,2H),1.80-1.84(m,1H),1.68(m,1H),1.27-1.48(m,5H),1.11(m,1H),1.01(m,1H),0.69(m,1H),0.46(m,1H),0.18-1.37(m,4H),0.11(m,2H);13C NMR(100MHz,CDCl3)δ181.3,82.8,58.0,50.8,46.4,46.0,45.0,44.6,35.2,34.9,28.2,26.3,25.7,17.5,11.5,9.6,9.2,7.9;ESI-HRMS m/z:304.2273[M+H]。
化合物28的合成方法:
将化合物27(38mg)溶于2mL二氯甲烷中,搅拌下缓慢加入0.05M稀盐酸直至pH为4,分出水相用二氯甲烷洗涤一次,水相冷冻干燥得化合物28(31mg),收率:73%。
分子式:C19H30NO2Cl
分子量:339
性状:白色固体
谱图数据:
1H-NMR(400MHz,D2O)δ4.42(m,1H),3.31-3.37(m,1H),3.21-3.25(m,1H),2.92(m,1H),2.82(s,3H),2.80(s,3H),2.11-2.26(m,2H),1.94(m,1H),1.79(m,1H),1.39-1.58(m,5H),1.23(m,1H),1.13(m,1H),0.69(m,1H),0.46(m,1H),0.18-1.33(m,4H),0.11(m,2H);13C NMR(100MHz,D2O)δ178.2,85.8,56.0,51.0,47.2,45.9,44.8,42.5,42.2,35.7,35.2,27.5,27.2,26.6,18.3,12.2,10.3,9.8,8.6;ESI-HRMSm/z:304.2271[M+H]
化合物29的合成:
向反应瓶中加入20mg化合物26,用1mL THF溶解,随后加入脯氨酸乙酯盐酸盐86mg,DBU 0.2mL,搅拌24小时,过柱纯化得化合物29(12mg).收率:39%。
分子式:C24H35NO4
分子量:401
性状:无色油状化合物
谱图数据:
1H NMR(400MHz,CDCl3):δ4.10-4.22(m,3H),3.31(dd,J=8.8,5.2Hz,1H),3.05(dd,J=13.2,5.2Hz,1H),2.91-2.96(m,1H),2.86(dd,J=13.2,3.6Hz,1H),2.37-2.49(m,3H),2.23-2.29(m,1H),1.96-2.11(m,3H),1.73-1.94(m,3H),1.57-1.69(m,3H),1.40-1.47(m,1H),1.17-1.36(m,6H),0.83-0.89(m,1H),0.70-0.75(m,1H),0.48-0.53(m,1H),0.40-0.45(m,1H),0.26-0.36(m,2H),0.17-0.24(m,2H);13C NMR(100MHz,CDCl3):δ178.3,174.4,84.3,66.6,60.6,53.9,51.9,51.7,47.5,47.2,44.7,36.6,36.2,29.3,29.1,27.6,27.0,24.0,18.8,14.5,12.8,10.6,10.1,9.1;ESI-HRMS m/z:402.2644[M+H]。
化合物(31)的合成方法:
取化合物26(51.6mg),二甲氨基吡啶(4mg)溶于2mL二氯甲烷,加入66mg苯硫酚,搅拌12小时,旋干,硅胶柱层析纯化得化合物31(68mg)。
1H NMR(400MHz,CDCl3)δ7.44-7.34(m,2H),7.30(dd,J=10.3,4.8Hz,2H),7.20(t,J=7.3Hz,1H),4.24-4.14(m,1H),3.47(dd,J=13.8,4.0Hz,1H),3.11(dd,J=13.8,7.0Hz,1H),2.52(ddd,J=11.2,7.0,4.1Hz,1H),2.30(ddd,J=20.7,10.6,4.2Hz,1H),2.21(dd,J=18.2,8.9Hz,1H),2.13-2.03(m,1H),1.96-1.86(m,1H),1.73-1.63(m,1H),1.61-1.47(m,2H),1.43(ddd,J=12.3,8.5,3.5Hz,1H),1.39-1.25(m,3H),0.89(dt,J=9.8,4.8Hz,1H),0.65(ddd,J=9.2,5.4,3.8Hz,1H),0.42(tt,J=13.4,4.6Hz,2H),0.36-0.30(m,1H),0.29-0.21(m,2H),0.18-0.08(m,1H);13C NMR(100MHz,CDCl3)δ177.1,136.3,130.0,129.6,127.0,84.2,77.92,52.6,48.3,47.4,47.1,35.9,35.4,33.8,29.7,27.7,27.0,18.8,12.8,11.4,9.3.
化合物(32)的合成方法:
化合物26(51.6mg),1,8-二氮杂二环[5.4.0]十一碳-7-烯(15.2mg)溶于0.5mL乙腈,加入37.3mg苄硫醇,搅拌24小时,旋干,硅胶柱层析纯化得化合物32(75mg)。1H NMR(400MHz,CDCl3)δ7.30-7.23(m,4H),7.22-7.17(m,1H),4.13(t,J=9.9Hz,1H),3.77-3.63(m,2H),2.80-2.66(m,2H),2.35(dt,J=10.5,5.1Hz,1H),2.28-2.14(m,2H),1.89(ddd,J=14.0,10.1,5.2Hz,2H),1.67-1.49(m,2H),1.48-1.38(m,2H),1.38-1.30(m,1H),1.30-1.17(m,2H),0.83(dt,J=9.7,4.9Hz,1H),0.63(ddd,J=9.4,5.4,4.0Hz,1H),0.48-0.40(m,1H),0.36(dt,J=8.9,4.5Hz,1H),0.33-0.25(m,1H),0.24-0.13(m,3H);13C NMR(100MHz,CDCl3)δ177.4,138.6,129.5,129.0,127.6,84.3,52.2,48.0,47.7,46.8,38.1,36.1,36.0,30.7,29.6,27.8,27.1,18.9,13.0,11.2,11.0,9.3.
化合物(33)的合成方法:
取化合物26(51.6mg)溶于1mL甲醇,加入321mg苄胺,搅拌24小时,旋干,硅胶柱层析纯化得中间体(76mg),将其溶于5mL甲醇,加入21mg甲基磺酸,搅拌十分钟后旋干得到化合物33(97mg)。
1H NMR(400MHz,MeOD)δ7.47(d,J=38.9Hz,5H),4.41(t,J=9.9Hz,1H),4.27(q,J=13.1Hz,2H),3.26(s,1H),2.93-2.79(m,1H),2.67(s,3H),2.39-2.28(m,1H),2.17(t,J=16.3Hz,1H),2.04(t,J=9.7Hz,1H),1.86(d,J=8.2Hz,1H),1.60(dd,J=17.1,7.0Hz,2H),1.45(d,J=35.5Hz,3H),1.30(dt,J=28.7,13.8Hz,3H),0.83(d,J=4.0Hz,1H),0.62(s,1H),0.48(d,J=3.8Hz,1H),0.41(d,J=3.9Hz,1H),0.30(d,J=4.0Hz,2H),0.21(s,2H);13C NMR(100MHz,MeOD)δ178.38,132.13,131.14,130.72,130.28,86.43,52.59,52.52,48.62,46.81,46.39,44.39,39.60,36.78,36.68,29.12,28.21,27.69,19.31,13.06,11.09,10.81,9.44.
化合物(34)的合成方法:
化合物26(103mg)溶于甲醇2mL,加入366mg氨基乙醇,搅拌24小时,硅胶柱层析纯化得胺加成中间体118mg,溶于5mL甲醇,加入甲基磺酸35.6mg,搅拌十分钟后旋干得到化合物34(153mg)。
1H NMR(400MHz,D2O)δ4.60(t,J=10.1Hz,1H),3.85(t,J=5.0Hz,2H),3.41-3.31(m,2H),3.29-3.18(m,2H),2.97-2.84(m,1H),2.78(s,3H),2.33(q,J=8.8Hz,2H),2.03(t,J=9.8Hz,1H),1.96-1.86(m,1H),1.67(dd,J=11.8,7.7Hz,1H),1.59-1.37(m,5H),1.35-1.23(m,1H),0.83-0.74(m,1H),0.55-0.38(m,3H),0.38-0.25(m,3H),0.24-0.19(m,1H);13C NMR(100MHz,D2O)δ178.39,85.67,56.28,50.62,49.68,46.96,45.74,45.67,43.11,38.58,35.95,35.62,27.66,27.22,26.69,18.27,12.28,10.14,9.49,8.72.
化合物(35)的合成方法:
取化合物26(103mg),二甲胺盐酸盐(489mg)溶于20mL二氯甲烷,加入1.66g碳酸钾,回流6小时,过滤,旋干,硅胶柱层析得到化合物27(110mg),取49mg化合物27溶于1mL甲醇,加入20.9mg二氯乙酸,搅拌10分钟,旋干得化合物35(69mg)。
1H NMR(400MHz,MeOD)δ5.91(s,1H),4.41(t,J=10.1Hz,1H),3.37(dd,J=13.3,9.2Hz,1H),3.22(dd,J=13.3,4.0Hz,1H),2.98-2.91(m,1H),2.85(s,6H),2.35(dd,J=17.4,9.1Hz,1H),2.25-2.14(m,1H),2.08-2.00(m,1H),1.98-1.88(m,1H),1.69-1.56(m,2H),1.56-1.44(m,3H),1.40-1.22(m,2H),0.85(dt,J=9.6,4.8Hz,1H),0.63(ddd,J=9.3,5.4,3.9Hz,1H),0.52-0.46(m,1H),0.45-0.39(m,1H),0.35-0.29(m,2H),0.21(td,J=9.5,5.2Hz,2H);13C NMR(100MHz,MeOD)δ178.45,170.46,86.32,70.47,57.60,52.72,48.77,47.52,44.52,43.69,36.93,36.85,29.11,28.36,27.85,19.45,13.22,11.23,10.93,9.58.
化合物(36)的合成方法:
取化合物26(50.0mg),溶于1mL无水甲醇,加入0.27mL哌啶,反应过夜,浓缩,硅胶柱层析纯化得胺加成中间体化合物50mg,将其溶于10mL甲醇,按等当量加入甲磺酸,旋干,得化合物36(63mg)。1H NMR(CD3OD,400MHz)δ0.13-0.18(m,2H),0.24-0.30(m,2H),0.33-0.38(m,1H),0.41-0.45(m,1H),0.55-0.59(m,1H),0.76-0.81(m,1H),0.92(d,J=6.5Hz,3H),1.19-1.66(m,9H),1.81-1.92(m,3H),1.96-2.01(m,1H),2.12-2.18(m,1H),2.26-2.31(m,1H),2.61(s,3H),2.92-3.02(m,3H),3.18-3.36(m,3H),3.53(t,J=13.6Hz,2H),4.37(t,J=10.1Hz,1H);13C NMR(CD3OD,100MHz)δ9.4,10.8,11.1,13.1,19.3,21.4,27.7,28.2,28.9,29.5,32.4,32.4,36.7,36.8,39.7,43.1,47.7,52.4,54.2,55.4,56.6,86.0,178.
化合物(37)的合成方法:
取化合物26(50.0mg),溶于1mL无水甲醇,加入0.30mL哌嗪,反应过夜,浓缩,硅胶柱层析纯化得胺加成中间体化合物51mg,将其溶于10mL甲醇,按等当量加入富马酸,旋干,得化合物37(62mg)。
1H NMR(CD3OD,400MHz)δ0.07-0.13(m,2H),0.17-0.22(m,2H),0.28-0.32(m,1H),0.36-0.40(m,1H),0.49-0.53(m,1H),0.72-0.76(m,1H),1.09-1.58(m,7H),1.84-1.95(m,2H),2.06-2.12(m,1H),2.18-2.25(m,1H),2.37-2.43(m,1H),2.52-2.69(m,6H),2.69(s,3H),3.10(brs,3H),3.16-3.17(m,1H),4.19(t,J=10.0Hz,1H),6.56(s,2H);13C NMR(CD3OD,100MHz)δ9.5,11.1,11.2,13.0,19.4,27.7,28.3,29.7,36.8,36.8,43.4,46.4,47.9,48.9,51.6,53.1,54.5,57.2,85.6,136,170.6,179.9.
化合物(38)的合成方法:
将化合物26(100mg,0.39mmol)溶于无水甲醇(20mL),加入吗啡啉(508mg,5.85mmol),室温搅拌过夜。硅胶柱层析法分离纯化得到产物(100mg,0.29mmol),将产物溶于无水甲醇(5mL),加入甲基磺酸(27.8mg,0.29mmol),将甲醇旋干得化合物38(127.8mg)。
1H NMR(400MHz,D2O)δ4.56(t,J=10.1Hz,1H),4.10(s,2H),3.92(m,J=17.5,12.5Hz,2H),3.67-3.46(m,3H),3.39(dd,J=13.8,3.3Hz,1H),3.35-3.25(m,2H),3.12-3.01(m,1H),2.77(s,3H),2.34(dt,J=15.8,7.9Hz,2H),2.05(t,J=9.9Hz,1H),1.93(s,1H),1.76-1.44(m,5H),1.43-1.33(m,1H),1.34-1.22(m,1H),0.88-0.74(m,1H),0.65-0.52(m,1H),0.52-0.18(m,6H);13C NMR(100MHz,D2O)δ178.38,85.98,63.53,55.30,50.93,48.80,47.00,45.84,41.70,38.45,35.16,34.68,27.34,26.77,26.31,17.94,11.76,9.99,9.71,8.11.
化合物(39)的合成方法:
将化合物26(50mg,0.195mmol)溶于无水甲醇(5mL),加入吡咯烷(208mg,2.93mmol),室温搅拌过夜。硅胶柱层析法分离纯化得到产物(52mg,0.16mmol),将产物溶于无水甲醇(5mL),加入甲基磺酸(15.2mg,0.16mmol),将甲醇旋干得化合物39(57.2mg)。
1H NMR(400MHz,D2O)δ4.44(t,J=10.0Hz,1H),3.79-3.58(m,2H),3.50(dd,J=13.4,9.5Hz,1H),3.35(dd,J=13.5,3.6Hz,1H),3.21-3.03(m,2H),2.90(dd,J=5.9,3.2Hz,1H),2.73(s,3H),2.33(d,J=8.5Hz,1H),2.24-2.08(m,3H),2.06-1.95(m,3H),1.92(s,1H),1.52(dt,J=24.8,8.8Hz,5H),1.33-1.15(m,2H),0.87-0.71(m,1H),0.65-0.53(m,1H),0.46-0.08(m,6H);13C NMR(100MHz,D2O)δ178.08,85.50,55.92,54.06,52.97,50.74,46.95,45.81,43.22,38.44,35.58,35.18,27.47,26.99,26.50,22.66,22.54,18.10,12.03,10.04,9.54,8.43.
化合物(40)的合成方法:
将化合物26(100mg,0.39mmol)溶于无水甲醇(2mL),加入N-甲基乙醇胺(438mg,5.85mmol),室温搅拌过夜。硅胶柱层析法分离纯化得到产物(100mg,0.30mmol),将产物溶于二氯甲烷(5mL),加入甲基磺酸(28.8mg,0.30mmol),将二氯甲烷旋干得化合物40(128.8mg)。
1H NMR(400MHz,CDCl3)δ4.36(t,J=9.9Hz,1H),3.92(s,2H),3.65-3.43(m,2H),3.34(s,2H),3.01(s,3H),2.98-2.83(m,1H),2.71(s,J=2.8Hz,3H),2.37-2.22(m,1H),2.19-2.10(m,2H),2.05-1.88(m,2H),1.57(m,J=29.9,18.2Hz,5H),1.31(d,J=10.9Hz,1H),1.21(s,1H),0.78(m,1H),0.59(m,1H),0.54-0.06(m,6H).
化合物(41)的合成方法:
将化合物26(50mg,0.195mmol)溶于无水甲醇(2mL),加入N-甲基丁胺(254mg,2.93mmol),室温搅拌过夜。硅胶柱层析法分离纯化得到胺加成中间体(50mg,0.14mmol),将其溶于二氯甲烷(5mL),加入甲基磺酸(13.9mg,0.14mmol),将二氯甲烷旋干得化合物41(63.9mg)。
1H NMR(400MHz,D2O)δ4.60(s,1H),3.59-3.07(m,4H),3.06-2.96(m,1H),2.90(s,3H),2.77(s,3H),2.32(dd,J=18.0,8.9Hz,2H),2.07-1.97(m,1H),1.94-1.81(m,1H),1.76-1.62(m,3H),1.61-1.51(m,2H),1.48(d,J=8.1Hz,1H),1.45-1.24(m,5H),0.91(t,J=7.4Hz,3H),0.83-0.76(m,1H),0.57-0.49(m,1H),0.45(ddd,J=17.8,8.7,4.7Hz,2H),0.38-0.24(m,3H),0.23-0.17(m,1H).
化合物42-45的合成方法:
在50mL圆底烧瓶中加入叠氮化三甲基硅烷(575mg,5.0mmol)和醋酸(300mg,5.0mmol),室温搅拌20分钟后,向该反应液中加入化合物26(258mg,1.0mmol)和催化量三乙胺(20.2mg,0.2mmol),升温至60℃搅拌直至反应完全(TLC检测)。把反应混合液倾入适量冰水中,用乙酸乙酯(50mL×3)萃取,接着依次用饱和碳酸氢钠溶液(20mL×3)和饱和食盐水(20mL×3)分别洗涤,无水硫酸钠干燥。抽滤浓缩后得叠氮化中间体。
取叠氮化中间体(0.25mmol),抗坏血酸钠(10mg,0.05mmol),五水硫酸铜(12mg,0.05mmol)于预先干燥的反应瓶中,抽真空,充氮气并封口。再加入混合溶剂(叔丁醇∶水=1∶1)5mL,5-己炔-1-醇(49mg,0.5mmol),室温搅拌直至原料反应完全(TLC检测)。把反应混合液倾入适量冰水中,用乙酸乙酯(10mL×3)萃取,接着用饱和食盐水洗涤三次,无水硫酸钠干燥,减压浓缩,硅胶柱层析纯化得化合物42(71mg)。
1H NMR(400MHz,CDCl3)δ7.40(s,1H),4.73-4.58(m,2H),4.23(t,J=10.0Hz,1H),3.65(t,J=6.3Hz,2H),2.73(t,J=7.4Hz,2H),2.63(dt,J=11.7,4.2Hz,1H),2.21(dd,J=18.0,9.0Hz,1H),1.96-1.89(m,1H),1.88-1.80(m,2H),1.79-1.69(m,3H),1.59(dt,J=16.7,7.6Hz,4H),1.52-1.44(m,2H),1.38-1.29(m,2H),1.27-1.19(m,1H),0.81(dd,J=9.2,4.8Hz,1H),0.65-0.54(m,1H),0.47-0.40(m,1H),0.40-0.35(m,1H),0.34-0.27(m,1H),0.23(m,3H);13C NMR(CDCl3,100MHz):176.1,149.0,122.3,84.9,62.4,52.0,48.6,47.9,47.5,44.6,36.0,35.5,32.4,28.6,27.6,27.0,25.9,25.6,18.6,12.8,11.1,10.9,9.1.
如上方法,制备得化合物43(70mg)。
1H NMR(400MHz,CDCl3)δ7.90(s,1H),7.81(d,J=7.6Hz,2H),7.39(t,J=7.4Hz,2H),7.29(dd,J=14.9,7.7Hz,1H),4.79-4.65(m,2H),4.20(t,J=9.9Hz,1H),2.68(dt,J=8.7,4.1Hz,1H),2.17(dd,J=17.9,8.9Hz,1H),2.07-1.92(m,1H),1.84(d,J=6.1Hz,1H),1.74(t,J=9.8Hz,1H),1.63-1.49(m,2H),1.42(dd,J=13.1,7.5Hz,2H),1.30(dt,J=20.0,9.9Hz,2H),1.24-1.15(m,1H),0.79(dd,J=17.1,12.5Hz,1H),0.61-0.51(m,1H),0.37(dd,J=20.4,4.1Hz,2H),0.29-0.13(m,4H);13C NMR(100MHz,CDCl3)δ175.54,148.13,130.37,128.88,128.29,125.73,120.84,84.53,51.46,48.24,47.47,47.20,44.21,35.62,35.21,28.27,27.24,26.61,18.29,12.51,10.69,10.42,8.70.
如上方法得化合物44(63mg)。
1H NMR(400MHz,CDCl3)δ8.21(s,1H),4.81-4.68(m,2H),4.28(dd,J=19.8,9.6Hz,1H),3.95(s,3H),2.76-2.65(m,1H),2.20(q,J=9.1Hz,1H),2.04-1.85(m,2H),1.74(t,J=9.9Hz,1H),1.65(dd,J=19.0,10.4Hz,1H),1.60-1.43(m,3H),1.42-1.32(m,2H),1.32-1.19(m,1H),0.81(dt,J=9.5,4.7Hz,1H),0.59(dt,J=9.2,4.5Hz,1H),0.49-0.36(m,2H),0.36-0.29(m,1H),0.30-0.20(m,3H);13C NMR(100MHz,CDCl3)δ175.09,160.87,140.15,128.87,84.49,52.19,51.58,48.00,47.55,47.51,44.20,35.51,34.96,28.18,27.18,26.54,18.23,12.42,10.76,10.53,8.64.
如上方法制备得化合物45(65mg)。
1H NMR(400MHz,CDCl3)δ7.58(d,J=33.3Hz,1H),5.07(d,J=4.9Hz,1H),4.78-4.60(m,2H),4.26(t,J=10.0Hz,1H),2.68(dt,J=9.5,4.4Hz,1H),2.23(dd,J=17.7,8.7Hz,1H),2.03-1.92(m,1H),1.89-1.73(m,2H),1.69-1.62(m,1H),1.62-1.54(m,4H),1.47(t,J=12.5Hz,2H),1.35(dd,J=20.3,10.3Hz,2H),1.30-1.18(m,2H),0.87-0.77(m,1H),0.67-0.57(m,1H),0.49-0.36(m,2H),0.36-0.29(m,1H),0.25(m,3H).
化合物(46)的合成方法:
取化合物26(167mg),氘代二甲胺盐酸盐(530mg)溶于15mL二氯甲烷,加入1.8g碳酸钾,搅拌24小时,过滤,旋干,过柱得到二甲胺基中间体(150mg),取49mg中间体,将其溶于10mL甲醇,加入18.7mg富马酸,搅拌10分钟,旋干得化合物46(67mg)。
1H NMR(400MHz,MeOD)δ6.70(s,2H),4.46(dd,J=18.9,8.7Hz,1H),3.46(dd,J=13.4,6.3Hz,1H),3.35-3.28(m,1H),2.95(d,J=14.0Hz,6H),2.38(dd,J=17.4,9.1Hz,1H),2.27-2.19(m,1H),2.11-2.03(m,1H),2.01-1.92(m,1H),1.67(ddd,J=18.7,11.2,5.0Hz,2H),1.60-1.48(m,3H),1.45-1.27(m,2H),0.88(dt,J=9.6,4.8Hz,1H),0.66(ddd,J=9.3,5.4,3.8Hz,1H),0.56-0.50(m,1H),0.45(dt,J=9.3,4.7Hz,1H),0.36(dt,J=11.9,4.7Hz,2H),0.29-0.20(m,2H);13C NMR(101MHz,MeOD)δ178.31,171.09,136.26,86.31,57.22,52.69,48.74,47.41,44.23,36.90,36.82,30.84,29.03,28.35,27.83,19.43,13.21,11.20,10.90,9.56.
化合物(47)的合成方法:
将小白菊内酯(566.2mg,1.99mmol)溶解于甲醇,加入对甲苯磺酸(687.5mg,4.0mmol)。反应体系放置于室温,搅拌过夜。反应混合液用Na2HPO4(568mg,4.0mmol)中和,然后用二氯甲烷萃取,收集有机层,并用Na2SO4干燥,过滤,再用旋转蒸发仪减压蒸馏有机溶剂,剩余的产物用硅胶柱纯化,得到化合物47。1H NMR(CDCl3,400MHz)δ6.12(d,J=2.8Hz,1H),5.44(s,1H),4.13(t,J=9.6Hz,1H),3.12(d,J=2.8Hz,3H),2.79(t,J=11.8Hz,2H),2.26(t,J=9.6Hz,1H),2.11-2.08(m,1H),1.98-1.62(m,6H),1.52-1.45(m,1H),1.31-1.37(m,4H),1.09(s,3H);13CNMR(CDCl3,100MHz)δ169.5,138.9,119.8,82.5,80.3,77.9,55.3,48.1,46.4,45.7,35.3,38.7,25.2,24.1,22.5,14.0。
化合物(48和49)的合成方法:
将小白菊内酯(1.03g,4.1mmol)溶液与丙酮∶水(95∶5)溶液中,加入amberlyst 15(12.88g),反应体系置于室温下反应,用TLC监测反应直至原料消失。待反应完全后,旋蒸除掉溶剂,剩余的物质溶于二氯甲烷中,并用Na2SO4干燥。过滤,旋蒸除掉溶剂,再用硅胶柱纯化得到产物48,49。
化合物48:1H NMR(CDCl3,400MHz)δ6.12(d,J=3.2Hz,1H),5.44(d,J=3.2Hz,1H),4.92(s,1H),4.88(s,1H),4.03-3.94(m,1H),2.93(dd,J=11.6,8.8Hz,1H),2.71-2.65(m,1H),2.61-2.56(m,1H),2.29(t,J=12Hz,1H),2.22-2.16(m,1H),1.85-1.74(m,3H),1.72-1.69(m,3H),1.34-1.26(m,3H),1.23(s,3H);13C NMR(CDCl3,100MHz)δ169.6,147.8,138.5,120.5,112.4,83.8,79.5,55.4,47.0,43.7,40.0,38.9,31.2,25.9,23.6。
化合物49:1H NMR(CDCl3,400MHz)δ6.24(d,J=3.6Hz,1H),5.56(d,J=3.2Hz,1H),4.26(dd,J=9.6,2.0Hz,1H),2.75-2.62(m,2H),2.40(t,J=12.4Hz,1H),2.20-2.14(m,1H),2.08(s,1H),2.03-1.96(m,2H),1.91-1.79(m,2H),1.76-1.60(m,2H),1.52-1.45(m,1H),1.36(s,3H),1.28(s,1H),1.25(s,3H);13C NMR(CDCl3,400MHz)δ169.6,138.4,120.4,82.7,80.0,74.7,55.2,49.6,47.1,39.3,25.2,24.9,24.1,23.4,20.6。
化合物50的制备方法:
将Martin’s sulfurane(382mg,0.57mmol)溶于2mL二氯甲烷中,在Ar保护的条件中,缓慢滴加化合物24的4mL二氯甲烷溶液。反应混合溶液逐渐变为黄色,搅拌24h,反应原料完全消失后,减压旋蒸除去溶剂,得到黄色油状物质,粗产品用硅胶柱纯化后,得到化合物50。1H NMR(CDCl3,400MHz)δ6.14(d,J=3.2Hz,1H),5.57(s,1H),5.41(d,J=3.2Hz,1H),4.00(t,J=10.4Hz,1H),2.93(br.d,J=10.8Hz,1H),2.77(br.d,J=17.6Hz,1H),2.27-2.12(m,3H),2.03(m,1H),1.97(br s,3H),1.84(br.d,J=14.0Hz,1H),1.48(m,1H),1.35(s,3H);13C NMR(CDCl3,400MHz)δ170.6,140.7,139.2,125.0,118.4,83.0,72.6,62.8,52.9,51.2,39.8,33.6,22.9,21.6,18.4.
实施例2:化合物1-50的抗类风湿性关节炎活性测试
在类风湿性关节炎药物领域,有很多文献报道药物成分对滑膜细胞分泌TNF-α,PGE2和IL-1β的影响,用动物实验的数据来考察药物治疗RA的效果。这些文献有:[1]鞠大宏,贾红伟,吴皓等,鹿瓜多肽注射液对C II诱导的免疫性关节炎大鼠血清TNF-α,IL-6以及C II抗体活性的影响,中国中医基础医学杂志,2003,9(11):17。[2]何金华,梁清华,张花生等,痹肿消汤对实验性关节炎大鼠血浆TNF-α的影响,湖南医科大学学报,2002,27(5):524.[3]黄清春,张声鹏,徐秋英,复方丹参对II型胶原蛋白诱导大鼠模型滑膜细胞分泌及肿瘤坏死因子的影响,现代康复,2001,5(10):54-55。[4]郑治刚,细胞因子及其检测方法和临床意义,陕西医学检验,2001,16(2):59。[5]周军,方素萍,齐云等,葛根汤对佐剂性关节炎大鼠关节液炎症介质的影响,中国实验方剂学杂志,2001,7(3):29。[6]马骥,卢秉久,朱晓明等,痛痹颗粒治疗类风湿性关节炎的药效学研究,中医药学刊,2001,19(6):734。[7]朱江,谢文利,晋玉璋等,栀子对类风湿性关节炎大鼠血清IL-1β和TNF-α的影响,中成药,2005,27(7):801。[8]黄清春,张声鹏,黄伟毅等,复方丹参注射液对CIA大鼠滑膜细胞IL-1βmRNA表达的影响,安徽中医学院学报,2002,21(5):39-41。
取本发明方法获得的化合物1-50,按文献[3]提供的方法进行模型制备、药物分组、大鼠滑膜细胞培养基上清的制备。
按文献[3]提供的方法考察测试化合物对滑膜细胞分泌TNF-α,PGE2的影响。按文献[8]提供的方法考察测试化合物对滑膜细胞分泌IL-1β的影响。
测试化合物1-50在剂量30mg/kg.2d下与正常对照、氯化钠组的实验数据如下:
表一 滑膜细胞培养上清中TNF-α的含量(x±s,ng/mL)
组别 |
TNF-α |
正常对照 |
0.38±0.027 |
氯化钠 |
0.74±0.098 |
化合物1 |
0.41±0.076 |
化合物2 |
0.45±0.052 |
化合物3 |
0.55±0.042 |
化合物4 |
0.56±0.064 |
化合物5 |
0.49±0.056 |
化合物6 |
0.61±0.072 |
化合物7 |
0.56±0.044 |
化合物8 |
0.58±0.053 |
化合物9 |
0.61±0.065 |
化合物10 |
0.54±0.055 |
化合物11 |
0.49±0.076 |
化合物12 |
0.66±0.043 |
化合物13 |
0.62±0.056 |
化合物14 |
0.48±0.055 |
化合物15 |
0.54±0.045 |
化合物16 |
0.57±0.073 |
化合物17 |
0.66±0.077 |
化合物18 |
0.64±0.072 |
化合物19 |
0.62±0.056 |
化合物20 |
0.58±0.045 |
化合物21 |
0.56±0.034 |
化合物22 |
0.61±0.082 |
化合物23 |
0.63±0.063 |
化合物24 |
0.54±0.057 |
化合物25 |
0.58±0.056 |
化合物26 |
0.59±0.051 |
化合物27 |
0.54±0.066 |
化合物28 |
0.47±0.056 |
化合物29 |
0.66±0.046 |
化合物30 |
0.65±0.057 |
化合物31 |
0.62±0.056 |
化合物32 |
0.64±0.044 |
化合物33 |
0.63±0.054 |
化合物34 |
0.54±0.067 |
化合物35 |
0.57±0.057 |
化合物36 |
0.66±0.045 |
化合物37 |
0.59±0.035 |
化合物38 |
0.67±0.045 |
化合物39 |
0.52±0.056 |
化合物40 |
0.48±0.078 |
化合物41 |
0.45±0.056 |
化合物42 |
0.58±0.055 |
化合物43 |
0.55±0.045 |
化合物44 |
0.54±0.071 |
化合物45 |
0.49±0.056 |
化合物46 |
0.56±0.034 |
化合物47 |
0.64±0.062 |
化合物48 |
0.62±0.054 |
化合物49 |
0.65±0.046 |
化合物50 |
0.52±0.064 |
表二 滑膜细胞培养上清中IL-1β的含量(x±s,ng/mL)
组别 |
IL-1β |
正常对照 |
0.16±0.034 |
氯化钠 |
0.49±0.136 |
化合物1 |
0.26±0.042 |
化合物2 |
0.28±0.045 |
化合物3 |
0.36±0.039 |
化合物4 |
0.42±0.064 |
化合物5 |
0.38±0.062 |
化合物6 |
0.36±0.054 |
化合物7 |
0.39±0.058 |
化合物8 |
0.41±0.051 |
化合物9 |
0.45±0.064 |
化合物10 |
0.42±0.069 |
化合物11 |
0.39±0.081 |
化合物12 |
0.38±0.069 |
化合物13 |
0.37±0.062 |
化合物14 |
0.36±0.054 |
化合物15 |
0.41±0.051 |
化合物16 |
0.34±0.039 |
化合物17 |
0.37±0.065 |
化合物18 |
0.39±0.029 |
化合物19 |
0.42±0.091 |
化合物20 |
0.34±0.056 |
化合物21 |
0.36±0.057 |
化合物22 |
0.34±0.038 |
化合物23 |
0.36±0.065 |
化合物24 |
0.37±0.051 |
化合物25 |
0.39±0.067 |
化合物26 |
0.42±0.052 |
化合物27 |
0.41±0.047 |
化合物28 |
0.31±0.067 |
化合物29 |
0.34±0.039 |
化合物30 |
0.38±0.049 |
化合物31 |
0.39±0.051 |
化合物32 |
0.37±0.047 |
化合物33 |
0.41±0.052 |
化合物34 |
0.35±0.059 |
化合物35 |
0.41±0.056 |
化合物36 |
0.29±0.064 |
化合物37 |
0.31±0.037 |
化合物38 |
0.35±0.062 |
化合物39 |
0.34±0.095 |
化合物40 |
0.31±0.061 |
化合物41 |
0.41±0.054 |
化合物42 |
0.36±0.059 |
化合物43 |
0.43±0.062 |
化合物44 |
0.35±0.069 |
化合物45 |
0.38±0.039 |
化合物46 |
0.36±0.067 |
化合物47 |
0.34±0.058 |
化合物48 |
0.38±0.046 |
化合物49 |
0.39±0.075 |
化合物50 |
0.34±0.061 |
表三 滑膜细胞培养上清中PGE2的含量(x±s,ng/mL)
组别 |
IL-1β |
正常对照 |
8.34±1.29 |
氯化钠 |
32.44±10.32 |
化合物1 |
15.24±1.77 |
化合物2 |
18.58±0.98 |
化合物3 |
26.96±1.59 |
化合物4 |
24.49±2.67 |
化合物5 |
25.67±4.23 |
化合物6 |
26.75±1.69 |
化合物7 |
22.82±2.67 |
化合物8 |
23.69±1.25 |
化合物9 |
27.92±2.67 |
化合物10 |
25.67±7.12 |
化合物11 |
26.38±3.15 |
化合物12 |
24.59±2.35 |
化合物13 |
25.61±0.98 |
化合物14 |
26.44±1.26 |
化合物15 |
24.39±2.38 |
化合物16 |
27.61±4.21 |
化合物17 |
28.95±3.24 |
化合物18 |
20.62±2.47 |
化合物19 |
26.45±5.21 |
化合物20 |
27.38±1.39 |
化合物21 |
19.59±3.25 |
化合物22 |
24.69±4.19 |
化合物23 |
21.71±2.32 |
化合物24 |
23.63±1.35 |
化合物25 |
26.48±2.57 |
化合物26 |
25.61±5.41 |
化合物27 |
24.75±2.66 |
化合物28 |
18.92±1.98 |
化合物29 |
27.64±2.37 |
化合物30 |
25.37±4.21 |
化合物31 |
24.41±1.65 |
化合物32 |
22.75±4.25 |
化合物33 |
26.64±2.67 |
化合物34 |
24.61±3.24 |
化合物35 |
21.54±1.32 |
化合物36 |
25.64±2.14 |
化合物37 |
27.94±2.69 |
化合物38 |
19.67±3.14 |
化合物39 |
28.35±2.15 |
化合物40 |
25.34±3.57 |
化合物41 |
26.63±1.59 |
化合物42 |
22.58±4.21 |
化合物43 |
24.64±3.95 |
化合物44 |
22.14±2.87 |
化合物45 |
23.26±3.49 |
化合物46 |
24.74±2.64 |
化合物47 |
27.95±3.67 |
化合物48 |
26.65±2.45 |
化合物49 |
24.25±1.94 |
化合物50 |
19.83±1.78 |
本发明以大鼠为人类RA动物模型,采用滑膜细胞原代培养的方法,观察了化合物1-50对滑膜细胞分泌TNF-α,PGE2和IL-1β的影响,结果表明,化合物1-50在大剂量组、小剂量组均能显著下调TNF-α,PGE2和IL-1β的含量,从而达到减轻致骨和软骨的破坏恢复关节活动功能,发挥治疗RA效果。
实施例3:化合物抑制癌症干细胞的活性测试
取新鲜或者冻存的(急性髓系白血病(AML,标记为CD34+/CD38+)、慢性髓系白血病(CML,Ph+/CD34+/CXCR4+)、慢性淋巴白血病(CLL,CD133+/CD19-/CD38-)、皮肤癌(CD34+)、乳腺癌(CD44+/CD24-/ESA+)、卵巢癌(CD44+/CD117+)、脑肿瘤(CD133+)、前列腺癌(CD44+/CD24-)、头颈鳞癌(CD44+)、喉癌(CD 133+)、胰腺癌(ESA+/CD44+/CD24+)、视网膜母细胞瘤(ABCG2/ALDH1)、儿童肝母细胞瘤(CD34+/THY1+/c-kit+)、肝癌(CD133+)、恶性黑色素瘤(CD133+)、大肠癌(EpCAMhigh/CD44+)、结肠腺癌(CD44high)、胶质瘤(ABCG-2/BCRP1)、消化道肿瘤(ABCG-2/BCRP1)、鼻咽癌(ABCG2)、脑胶质瘤(Dlk-1/Pref-1)、胃癌(CD45+)、肺腺癌(Sca-1/CD45-/Pecam-/CD34+)、肺癌(CD133+CD34+CD44+))病人临床标本。以急性髓系白血病为例,用Ficoll进行密度梯度离心,取出中间的单个核细胞群,离心后用无血清的IMDM重悬。将细胞调成1x106/mL,铺入24孔板,每孔1mL,用相应浓度的待选化合物处理18个小时,18个小时后,收细胞,1500转8分钟离心,沉淀用100微升重悬,标记白血病干细胞的相关抗体,室温避光,孵育30分钟,30分钟后1500转离心,洗掉未结合的抗体CD34+CD38+,用100微升1xBinding Buffer重悬,分别加入5微升Annexin V-FITC和PI,室温避光,孵育十五分钟,后加入200微升1xBinding Buffer,在一个小时内用流式细胞仪检测标记的细胞凋亡。
表四 加入测试化合物后,癌细胞、癌症干细胞和正常细胞的存活率(%)
实施例4:注射液
实施例1所制备化合物1-50用少量的DMSO溶解后,按常规加注射用水,精滤,灌封灭菌制成注射液。
实施例5:片剂
实施例1所制备化合物1-50与赋形剂按照重量比为5∶1的比例加入赋形剂,制粒压片,得片剂。
实施例6:胶囊
实施例1所制备化合物1-50与赋形剂按照重量比为5∶1的比例加入赋形剂,制成胶囊。
本发明的化合物、用途和方法已经通过具体的实施例进行了描述。本领域技术人员可以借鉴本发明的内容适当改变原料、工艺条件等环节来实现相应的其它目的,其相关改变都没有脱离本发明的内容,所有类似的替换和改动对于本领域技术人员来说是显而易见的,都被视为包括在本发明的范围之内。