CN102850301A - 水溶性紫杉醇衍生物及其药物组合物及其医药用途 - Google Patents
水溶性紫杉醇衍生物及其药物组合物及其医药用途 Download PDFInfo
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Abstract
Description
技术领域
本发明涉及紫杉醇类抗肿瘤药物紫杉醇及多烯紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐。
背景技术
紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇是目前临床最常用的抗肿瘤药物,其化学结构如下所示:
紫杉醇类药物是具有微管蛋白抑制活性的抗肿瘤药物。紫杉醇及其衍生物多西紫杉醇能促使微管蛋白聚集成微管,并结合到微管上抑制微管的解聚,从而使细胞有丝分裂中止。紫杉醇和多西紫杉醇对卵巢癌、乳腺癌、非小细胞性肺癌、头颈部恶性肿瘤等有显著的疗效,并且能够与其它抗肿瘤药物顺铂联合用药以提高疗效,成为应用最广泛的抗肿瘤药物。但是紫杉醇和多西紫杉醇的水溶解度很差。
发明内容
本发明提供式I所示的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐:
式I中,n为1-4的整数;RR代表天然或非天然的L型或D型氨基酸的侧链,包括H,-CH3,-CH(CH3)2,-CH2CH(CH3)2,-CH(CH3)(CH2CH3),-CH2OH,-CH(OH)(CH3),-CH2SH,-CH2CH2SCH3,-CH2COOH,-CH2CONH2,-CH2CH2COOH,-CH2CH2CONH2,-(CH2)3NHC(NH)(NH2),-(CH2)4NH2;
优选地,本发明提供紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐,结构如式Ia所示:
式Ia中,n为1-4的整数;R代表天然或非天然的L型或D型氨基酸的侧链,包括H,-CH3,-CH(CH3)2,-CH2CH(CH3)2,-CH(CH3)(CH2CH3),-CH2OH,-CH(OH)(CH3),-CH2SH,-CH2CH2SCH3,-CH2COOH,-CH2CONH2,-CH2CH2COOH,-CH2CH2CONH2,-(CH2)3NHC(NH)(NH2),-(CH2)4NH2。
优选地,本发明还提供多烯紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐,结构如式Ib所示:
式Ib中,n为1-4的整数;R代表天然或非天然的L型或D型氨基酸的侧链,包括H,-CH3,-CH(CH3)2,-CH2CH(CH3)2,-CH(CH3)(CH2CH3),-CH2OH,-CH(OH)(CH3),-CH2SH,-CH2CH2SCH3,-CH2COOH,-CH2CONH2,-CH2CH2COOH,-CH2CH2CONH2,-(CH2)3NHC(NH)(NH2),-(CH2)4NH2。
本发明还提供式I所示的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物的非毒性药学上可接受的盐,如无机金属离子的钠盐、钾盐或有机胺的盐。
本发明还提供含有式I所示的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐作为活性成分的药物组合物。
本发明最后还提供式I所示的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐,以及含有式I所示的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐作为活性成分的药物组合物作为抗肿瘤药物的用途。
具体实施方式
以紫杉醇的水溶性衍生物Ia为例,目标化合物可以通过如下合成路线制备:
二元酸酐如丙二酸酐、琥珀酸酐或戊二酸酐与紫杉醇反应,在吡啶的催化下生成紫杉醇-2′-二元酸的单酯衍生物,然后单酯衍生物的羧基再与侧链保护的氨基酸反应,最后在10%的钯碳催化氢化下,脱去侧链及羧基保护基,得到目标产物。
通过下面的实施例可以对本发明进行进一步的描述,然而,本发明的范围并不限于下述实施例。本领域的专业人员能够理解,在不背离本发明的精神和范围的前提下,可以对本发明进行各种变化和修饰。
实施例1N-(紫杉醇-2′-氧羰基丙酰)-L-天冬氨酸(Ia-1)的制备
1.1紫杉醇-2′-琥珀酸单酯(IIa-1)的制备
将1200mg(1.404mmol)紫杉醇,168.8mg(1.688mmol)琥珀酸酐,36mg(0.28mmol)DMAP溶于40ml无水吡啶中。室温搅拌3小时。旋干溶剂,残留物用少量二氯甲烷溶解,柱层析(二氯甲烷∶甲醇=10∶1)得IIa-1的白色粉末状固体1336mg,产率99.7%。MS(ESI)m/z:954.8(M+H+),1H-NMR(CDCl3):δ1.12(s,3H),1.22(s,3H),1.67(s,3H),1.90(brs.,4H),2.15(brs.,3H),2.36-2.60(m,10H),3.79(d,J=7.0Hz,1H),4.18(d,J=8.4Hz,1H),4.28(d,J=8.2Hz,1H),4.42(dd,J1=12.4Hz,J 2=6.8Hz,1H),4.96(d,J=8.6Hz,1H),5.50(d,J=3.1Hz,1H),5.68(d,J=6.9Hz,1H),5.98(dd,J1=12.4Hz,J2=3Hz,1H),6.23(t,J=9.1Hz,1H),6.29(s,1H),7.20(d,J=9.2Hz,1H),7.30-7.40(m,7H),7.43-7.49(m,3H),7.59-7.62(m,1H),7.74(d,J=7.2Hz,2H),8.12(d,J=7.2Hz,2H)
1.2N-(紫杉醇-2′-氧羰基丙酰)-L-天冬氨酸苄酯(IIIa-1)的制备
25ml烧瓶中加入IIa-1 400mg,加入无水二氯甲烷10ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Asp(OBzl)-OBzl·TosOH 225mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-1的白色粉末状固体365mg。产率71.3%MS(ESI)m/z:1249.8(M+H+),1H-NMR(CDCl3):δ1.12(s,3H),1.13(s,3H),1.90(brs.3H),2.11(m,1H),2.22(s,3H),2.30(m,1H),2.41(s,3H),2.51(m,4H),2.74-2.83(m,2H),3.22(q,J=17.6Hz,1H),3.84(d,J=7.0Hz,1H),4.00(m,1H),4.21(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J2=6.8Hz,1H),4.72(m,1H),4.98(d,J=8.6Hz,1H),5.05-5.12(ss,4H),5.48(d,J=2.8Hz,1H),5.71(d,J=6.8Hz,1H),5.93(dd,J1=12.4Hz,J2=3Hz,1H),6.03(d,J=8.8Hz,1H),6.24(t,J=9.1Hz,1H),6.30(s,1H),6.51(d,J=8.0Hz,1H),7.17-7.6(m,26H),8.14(d,J=6.8Hz,2H)
1.3N-(紫杉醇-2′-氧羰基丙酰)-L-天冬氨酸(Ia-1)的制备
将IIIa-1 365mg加入30ml甲醇溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除滤剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得Ia-1的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1069.9(M-H+),1H-NMR(400M Hz DMSO-d6)δppm:0.99(3H,s,17-H),1.04(3H,s,16-H),1.498(4H,brs),1.77(4H,brs),2.10(3H,s,10-OAc),2.247(3H,s,4-OAc),2.50-2.70(6H,m,-CH2COOR),3.59(1H,d,J=7.2Hz,3-H),3.99-4.03(2H,q,J=12.4Hz,20-H),4.12(1H,m,7-H),4.48(1H,d,J=4.4Hz,6″-H),4.66(1H,s,5-H),4.93(2H,t,J=6.8Hz),5.37(1H,d,J=8.4Hz,2’-H),5.41(1H,d,J=6.8Hz,2-H),5.6(1H,q,J=5.5Hz,3’-H),5.83(1H,t,J=9.0Hz,13-H),6.29(1H,s,10-H),7.10-8.29(16H,m),9.24(1H,d,J=8.8Hz,NH).
实施例2N-(紫杉醇-2′-氧羰基丙酰)-L-谷氨酸(Ia-2)的制备
2.1N-(紫杉醇-2′-氧羰基丙酰)-L-谷氨酸苄酯(IIIa-2)的制备
25ml烧瓶中加入IIa-1 400mg,加入无水二氯甲烷溶液20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Glu(OBzl)-OBzl·TosOH 230mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-2的白色粉末状固体380mg。产率66.4%.MS(ESI)m/z:1263.8(M+H+),1H-NMR(CDCl3):δ1.13-1.25(m,6H),1.67(s,3H),1.90(brs.5H),2.1(s,3H),2.3(m,3H),2.52(brs,5H),2.74(m,2H),3.84(d,J=6.0Hz,1H),4.21(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J2=6.8Hz,1H),4.50(m,1H),4.98(d,J=8.6Hz,1H),5.05-5.18(ss,4H),5.455(d,J=2.8Hz,1H),5.67(d,J=6.8Hz,1H),5.95(dd,J1=12.4Hz,J2=3Hz,1H),6.03(d,J=8.8Hz,1H),6.24(t,J=9.1Hz,1H),6.30(s,1H),6.44(t,J=5.6Hz,1H),7.26-7.8(m,26H),8.14(d,J=6.8Hz,2H).
2.2N-(紫杉醇-2′-氧羰基丙酰)-L-谷氨酸(Ia-2)的制备
将IIIa-2380mg用甲醇30ml溶解,加入10%Pd/C 30mg,催化氢化反应5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除滤剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得Ia-2的白色粉末状固体330mg,产率100%。MS(ESI)m/z:1083.9(M-H+),1H-NMR(400M Hz,DMSO-d6)δppm:1.00(3H,s,17-H),1.02(3H,s,16-H),1.499-1.75(9H,brs),2.11(3H,s,10-OAc),2.24-2.29(5H,m,4-OAc,1″′-CH2),2.55-2.60(2H,m,2″′-CH2COOH),2.73(3H,s,2″-H),3.57(1H,d,J=7.19Hz,3-H),3.99-4.01(2H,q,J=12.4Hz,20-H2),4.12(2H,m,7-H,2″-H),4.66(1H,s,5-H),4.92(2H,d,J=11.2HZ,),5.36(1H,d,J=5.5Hz,2’-H),5.41(1H,d,J=6.8Hz,3’-H),5.47(1H,d,J=7.2Hz,2-H),5.83(1H,t,J=9.0Hz,13-H),6.29(1H,s,10-H),7.19-8.13(16H,m),9.25(1H,d,J=8.8Hz,NH),12.781(br-s,-COOH)
实施例3N-(紫杉醇-2′-氧羰基丙酰)-L-丙氨酸(Ia-3)的制备
3.1N-(紫杉醇-2′-氧羰基丙酰)-L-丙氨酸苄酯(IIIa-3)的制备
25ml烧瓶中加入IIa-1400mg,加入无水二氯甲烷20ml溶液使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Ala-OBzl·TosOH 179mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-3白色粉末状固体290mg。产率62.0%。MS(ESI)m/z:1115.9(M+H+),1H-NMR(400MHz,DMSO-d6)δppm:0.99(3H,s,17-H),1.03(3H,s,16-H),1.16(3H,d,J=6.8,1″′-H),1.49(4H,s,19-H),1.63(3H,m),2.10(3H,s,10-OAc),2.24(3H,s,4-OAc),2.42-2.50(4H,m),2.60(2H,t,J=7.2Hz,4″-CH2COO),3.59(1H,d,J=7.2Hz,3-H),3.95-4.05(2H,m,20-H2,7″-H),4.12(1H,m,7-H),4.2(1H,t,J=7.6Hz),4.65(1H,s,5-H),4.93(2H,m),5.1(2H,s,-COOCH2Ph),5.345(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H),5.53(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(21H,m),9.24(1H,d,J=8.8Hz,NH).
3.2N-(紫杉醇-2′-氧羰基丙酰)-L-丙氨酸(Ia-3)的制备
将IIIa-3290mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-3的白色粉末状固体274mg,产率100%。MS(ESI)m/z:1023.9(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:0.99(3H,s,17-H),1.03(3H,s,16-H),1.16(3H,d,J=6.8,1″′-H),2.3(1H,brs),1.49(4H,brs),1.62(2H,t,J=12.4Hz),1.75(4H,brs),2.10(3H,s,10-OAc),2.23(3H,s,4-OAc),2.42(2H,t,J=7.2Hz,2″-CH2COO),2.60(2H,t,J=7.2Hz,4″-CH2COO),3.59(1H,d,J=7.2Hz,3-H),3.95-4.05(2H,m,20-H,7″-H),4.12(1H,m,7-H),4.68(1H,s,5-H),4.93(2H,d,J=6.8Hz),5.35(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H)5.53(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(16H,m),9.31(1H,d,J=8.4Hz,NH).
实施例4N-(紫杉醇-2′-氧羰基丙酰)-L-亮氨酸(Ia-4)的制备
4.1N-(紫杉醇-2′-氧羰基丙酰)-L-亮氨酸苄酯(IIIa-4)的制备
25ml烧瓶中加入IIa-1 400mg,加入无水二氯甲烷30ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Leu-OBzl·TosOH 174mg(MW=393.5,1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-4的白色粉末状固体298mg。产率61.4%。MS(ESI)m/z:1157.9(M+H+),1H-NMR(CDCl3):δ0.82-0.84(m,6H),1.12(s,3H),1.22(s,3H),1.49-2.0(brs,12H),2.2-2.7(m,9H),3.79(d,J=7.0Hz,1H),4.19(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J2=6.8Hz,1H),4.58(m,1H),4.96(d,J=8.6Hz,1H),5.10(m,2H),5.465(d,J=3.6Hz,1H),5.68(d,J=7.6Hz,1H),5.98(dd,J1=12.4Hz,J2=3Hz,1H),6.09(d,J=8.8Hz,1H),6.23(t,J=9.1Hz,1H),6.28(s,1H),7.17-7.8(m,21H)8.14(d,J=6.8Hz,2H).
4.2N-(紫杉醇-2′-氧羰基丙酰)-L-亮氨酸(Ia-4)的制备
将IIIa-4298mg用甲醇30ml溶解,加入10%Pd/C 30mg,催化氢化反应5小时,反应完成后用微孔滤过膜(有机相)滤除Pd/C,蒸除溶剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得Ia-4的白色粉末状固体274mg,产率100%。MS(ESI)m/z:1066.1(M-H+),1H-NMR(400M Hz,DMSO-d6)δppm:0.79-0.86(6H,m,2″′-CH3,2″′-CH3),1.00(3H,s,17-H),1.03(3H,s,16-H),1.5(6H,brs),1.79(3H,s),2.10(3H,s,10-OAc),2.24(2H,m,2″-H),2.26(3H,s,4-OAc),2.41(2H,t,J=6.2Hz,4″-CH2COO),2.59(2H,t,J=6.0Hz,2″-CH2COO),3.58(1H,d,3-H),3.95-4.01(2H,q,J=12.4Hz,20-H),4.12-4.29(2H,m,7″-H,7-H),4.66(1H,s,5-H),4.91(2H,d,J=9.8Hz),5.34(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H),5.50(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(16H,m),9.31(1H,d,J=8.4Hz,NH).
实施例5N-(紫杉醇-2′-氧羰基丙酰)-L-异亮氨酸(Ia-5)的制备
5.1N-(紫杉醇-2′-氧羰基丙酰)-L-异亮氨酸苄酯(IIIa-5)的制备
25ml烧瓶中加入IIa-1 400mg,加入无水二氯甲烷溶液20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Ile-OBzl·TosOH 174mg(MW=393.5,1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-5白色粉末状固体301mg。产率61.5%。MS(ESI)m/z:1157.9(M+H+),1H-NMR(CDCl3):δ0.82-0.84(m,6H),1.12(s,3H),1.22(s,3H),1.67(s,3H),1.90(brs.7H),2.2(brs.,3H),2.4-2.7(m,8H),3.79(d,J=7.0Hz,1H),4.19(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J2=6.8Hz,1H),4.58(m,1H),4.96(d,J=8.6Hz,1H),5.05-5.17(m,2H),5.465(d,J=3.6Hz,1H),5.68(d,J=7.6Hz,1H),5.98(dd,J1=12.4Hz,J2=3Hz,1H),6.09(d,J=8.8Hz,1H),6.23(t,J=9.1Hz,1H),6.28(s,1H),7.17-7.8(m,21H)8.13(d,J=6.8Hz,2H)
5.2N-(紫杉醇-2′-氧羰基丙酰)-L-异亮氨酸(Ia-5)的制备
将IIIa-5 301mg用甲醇30ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除溶剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-5的白色粉末状固体279mg,产率100%。MS(ESI)m/z:1066.1(M-H+),1H-NMR(400M Hz,DMSO-d6)δppm:0.796-0.887(12H,m,3″′-H,2″′-CH3,17-H,16-H),1.497(3H,s,19-H),1.771(3H,s,18-H3),1.60-1.73(4H,m,2″′-H,2″-H),2.10(3H,s,10-OAc),2.23(3H,s,4-OAc),2.46(2H,t,J=7.2Hz,2″-CH2COO),2.59(2H,t,J=7.2Hz,4″-CH2COO),3.60(1H,m,3-H),4.01-4.14(4H,m,20-H,7″-H,7-H),4.675(1H,s,5-H),4.91(2H,d,J=9.8Hz),5.33(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H)5.51(1H,t,J=8.4Hz,3’-H),5.63(1H,d,7.6Hz).5.83(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(16H,m),9.28(1H,d,J=8.4Hz,NH).
实施例6N-(紫杉醇-2′-氧羰基丙酰)-D-天冬氨酸(Ia-6)的制备
6.1N-(紫杉醇-2′-氧羰基丙酰)-D-天冬氨酸苄酯(IIIa-6)的制备
25ml烧瓶中加入IIa-1 400mg,加入无水二氯甲烷溶液30ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-D-Asp(OBzl)-OBzl·TosOH 225mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-6的白色粉末状固体370mg。产率71.5%,MS(ESI)m/z:1249.9(M+H+),1H-NMR(CDCl3):δ1.12(s,3H),1.25(s,3H),1.69(brs,3H),1.90(brs.4H),2.22(s,3H),2.41(s,3H),2.51(m,2H),2.74-2.83(m,3H),3.84(d,J=7.0Hz,1H),4.00(m,1H),4.21(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J2=6.8Hz,1H),4.72(m,1H),4.98(d,J=8.6Hz,1H),5.05-5.12(ss,4H),5.48(d,J=2.8Hz,1H),5.71(d,J=6.8Hz,1H),5.93(dd,J1=12.4Hz,J2=3Hz,1H),6.03(d,J=8.8Hz,1H),6.24(t,J=9.1Hz,1H),6.30(s,1H),6.51(d,J=8.0Hz,1H),7.17-7.6(m,26H),8.14(d,J=6.8Hz,2H).
6.2N-(紫杉醇-2′-氧羰基丙酰)-D-天冬氨酸(Ia-6)的制备
将IIIa-6365mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-6白色粉末状固体315mg,产率100%。MS(ESI)m/z:1067.9(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:0.99(3H,s,17-H),1.04(3H,s,16-H),1.498(4H,brs),1.77(3H,s),2.10(3H,s,10-OAc),2.25(3H,s,4-OAc),2.41-2.66(6H,m,-CH2COOR),3.16(1H,s),3.59(1H,d,J=7.2Hz,3-H),3.99-4.03(2H,q,J=12.4Hz,20-H),4.11(1H,m,7-H),4.41(1H,d,J=4.4Hz,6″-H),4.67(1H,s,5-H),4.93(2H,m),5.36(1H,d,J=8.4Hz,2’-H),5.42(1H,d,J=6.8Hz,2-H),5.50(1H,q,J=5.5Hz,3’-H),5.83(1H,t,J=9.0Hz,13-H),6.29(1H,s,10-H),7.10-8.29(16H,m),9.25(1H,d,J=8.8Hz,NH).
实施例7N-(紫杉醇-2′-氧羰基丁酰)-L-天冬氨酸(Ia-7)的制备
7.1紫杉醇-2′-戊二酸单酯(IIa-2)的制备
将2000mg(2.35mmol)紫杉醇,308mg(2.7mmol)戊二酸酐,60mg(0.47mmol)DMAP溶于40ml无水吡啶中。室温搅拌反应3小时。旋干溶剂,残留物用少量二氯甲烷溶解,柱层析(二氯甲烷∶甲醇=10∶1)得IIa-2的白色粉末状固体2202mg,产率99.1%。MS(ESI)m/z:968.7(M+H+),1H-NMR(400Mz,CDCl3):δppm 1.13(s,3H),1.24(s,3H),1.66(s,3H),1.90(brs.,5H),2.22-2.60(m,13H),3.81(d,J=7.0Hz,1H),4.18(d,J=8.4Hz,1H),4.28(d,J=8.2Hz,1H),4.42(dd,J1=12.4Hz,J2=6.8Hz,1H),4.98(d,J=8.6Hz,1H),5.50(d,J=3.1Hz,1H),5.68(d,J=6.9Hz,1H),6.0(dd,J1=12.4Hz,J2=3Hz,1H),6.24(t,J=9.2Hz,1H),6.29(m,1H),7.20-8.12(m,16H).
7.2N-(紫杉醇-2′-氧羰基丁酰基)-L-天冬氨酸苄酯(IIIa-7)的制备
25ml烧瓶中加入IIa-2 400mg,加入无水二氯甲烷溶液20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Asp(OBzl)-OBzl·TosOH 200mg(MW=485.6,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-7的白色粉末状固体355mg。产率68.1%。MS(ESI)m/z:1263.8(M+H+),1H-NMR(400Mz,CDCl3):δppm 1.10-1.23(m,6H),1.69(brs,4H),1.90(brs.5H),2.1-2.22(m,5H),2.52(brs,4H),3.47(m,1H),3.84(d,J=7.0Hz,1H),3.86(s,1H),4.21(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J 2=6.8Hz,1H),4.75(m,1H),4.98(d,J=8.6Hz,1H),5.05-5.12(ss,4H),5.52(d,J=2.8Hz,1H),5.69(d,J=6.8Hz,1H),6.01(dd,J1=12.4Hz,J2=3Hz,1H),6.03(d,J=8.8Hz,1H),6.26(t,J=9.1Hz,1H),6.30(s,1H),6.4(t,J=5.6Hz,1H),6.57(d,J=8.0Hz,1H),7.23-7.74(m,26H),8.00(s,1H),8.14(d,J=6.8Hz,2H).
7.3N-(紫杉醇-2′-氧羰基丁酰)-L-天冬氨酸(Ia-7)的制备
将IIIa-7365mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-7的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1084.0(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:1.0(3H,s,17-H),1.04(3H,s,16-H),1.50-1.78(9H,m),2.10-2.17(5H,m),2.25(3H,s,4-OAc),2.39-2.60(6H,m,-CH2COO),3.58(1H,d,J=7.2Hz,3-H),3.99-4.01(2H,q,J=12.4Hz,20-H),4.12(1H,m,7-H),4.35(1H,d,J=4.4Hz,6″-H),4.66(1H,s,5-H),4.93(2H,t,J=6.8Hz),5.37(1H,d,J=8.4Hz,2’-H),5.41(1H,d,J=6.8Hz,2-H),5.5(1H,q,J=5.5Hz,3’-H),5.62(1H,d,J=7.6Hz),5.81(1H,t,J=9.0Hz,13-H),6.30(1H,s,10-H),7.10-8.29(16H,m),9.265(1H,d,J=8.8Hz,NH).
实施例8N-(紫杉醇-2′-氧羰基丁酰)-L-谷氨酸(Ia-8)的制备
8.1N-(紫杉醇-2′-氧羰基丁酰)-L-谷氨酸苄酯(IIIa-8))的制备
25ml烧瓶中加入IIa-2400mg,加入无水二氯甲烷溶液20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Glu(OBzl)-OBzl·TosOH 207mg(MW=499.6,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-8的白色粉末状固体410mg。产率77.8%。MS(ESI)m/z:1277.8(M+H+),1H-NMR(400Mz,CDCl3);δ1.13-1.25(m,6H),1.69(s,4H),1.90(brs.8H),2.19-2.56(m,15H),3.84(d,J=7.0Hz,1H),4.22(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.48(dd,J1=12.4Hz,J2=6.8Hz,1H),4.98(d,J=8.6Hz,1H),5.05-5.12(ss,4H),5.52(d,J=2.8Hz,1H),5.69(d,J=6.8Hz,1H),6.02(dd,J1=12.4Hz,J2=3Hz,1H),6.26(t,J=9.1Hz,1H),6.30(s,1H),7.17-7.6(m,26H),8.14(d,J=6.8Hz,2H).
8.2N-(紫杉醇-2′-氧羰基丁酰)-L-谷氨酸(Ia-8)的制备
将IIIa-8410mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-8的白色粉末状固体348mg,产率100%。MS(ESI)m/z:1097.9(M+H+),1H-NMR(400M Hz DMSO-d6)δppm:0.99(3H,s,17-H),1.02(3H,s,16-H),1.49(4H,s,19-H),1.78(10H,brs),2.11(3H,s,10-OAc),2.27(3H,s,4-OAc),2.28-2.509(6H,m,-CH2COOR),3.58(1H,d,J=7.2Hz,3-H),4.03-4.17(4H,q,J=12.4Hz,20-H,7-H,6″-H),4.66(1H,s,5-H),4.91(2H,t,J=6.8Hz),5.34(1H,d,J=8.4Hz,2’-H),5.41(1H,d,J=6.8Hz,2-H),5.52(1H,q,J=5.5Hz,3’-H),5.81(1H,t,J=9.0Hz,13-H),6.29(1H,s,10-H),7.10-8.29(16H,m),9.25(1H,d,J=8.8Hz,NH),12.810(br-s,-COOH).
实施例9N-(紫杉醇-2′-氧羰基丁酰)-L-丙氨酸(Ia-9)的制备
9.1N-(紫杉醇-2′-氧羰基丁酰)-L-丙氨酸苄酯(IIIa-9)的制备
25ml烧瓶中加入IIa-2400mg,加入无水二氯甲烷溶液20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Ala(OBzl)-OBzl·HCl 160mg(MW=351.4,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml ×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-9的白色粉末状固体370mg。产率79.3%。MS(ESI)m/z:1130.0(M+H+),1H-NMR(400MHz,DMSO-d6δppm:0.99(3H,s,17-H),1.03(3H,s,16-H),1.26(3H,d,J=7.6Hz),1.47-1.83(10H,m,19-H,18-H),2.10(3H,s,10-OAc),2.15(2H,t,J=7.2Hz),2.245(3H,s,4-OAc),2.42(2H,m,-CH2COOR),3.59(1H,d,J=7.2Hz,3-H),4.01(2H,q,J=12.4Hz,20-H),4.10(1H,m,7-H),4.27(1H,t,J=6.8Hz),4.62(1H,s,5-H),4.92(2H,d,J=6.8Hz),5.35(1H,d,J=8.4Hz,2’-H),5.40(1H,d,J=6.8Hz,2-H),5.50(1H,t,J=9.6Hz,3’-H),5.82(1H,t,J=8.4Hz,13-H),6.30(1H,s,10-H),7.18-7.99(21H,m),9.21(1H,d,J=8.8Hz,NH).
9.2N-(紫杉醇-2′-氧羰基丁酰)-L-丙氨酸(Ia-9)的制备
将IIIa-9370mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得白色粉末状固体Ia-9340mg,产率100%。MS(ESI)m/z:1038.1(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:0.99(3H,s,17-H),1.04(3H,s,16-H),1.496-1.84(14H,m),2.10(3H,s,10-OAc),2.25(3H,s,4-OAc),2.40-2.50(6H,m,-CH2COOR),3.59(1H,d,J=7.2Hz,3-H),3.95-4.01(2H,q,J=12.4Hz,20-H),4.12(1H,m,7-H),4.68(1H,s,5-H),4.92(2H,d,J=6.8Hz),5.35(1H,d,J=8.4Hz,2’-H),5.40(1H,d,J=6.8Hz,2-H),5.53(1H,q,J=5.5Hz,3’-H),5.82(1H,t,J=9.0Hz,13-H),6.30(1H,s,10-H),7.18-7.99(16H,m),9.31(1H,d,J=8.8Hz,NH).
实施例10N-(紫杉醇-2′-氧羰基丁酰)-L-甘氨酸(Ia-10)的制备
10.1N-(紫杉醇-2′-氧羰基丁酰)-L-甘氨酸苄酯(IIIa-10)的制备
25ml烧瓶中加入IIa-2 400mg,加入无水二氯甲烷溶液20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Gly(OBzl)-OBzl·TosOH 154mg(MW=337.4,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-10白色粉末状固体340mg。产率73.9%。MS(ESI)m/z:1115.9(M+H+),1H-NMR(CDCl3):δ1.15(s,3H),1.17(s,3H),1.69(brs,4H),1.90(brs.6H),2.2-2.6(m,8H),3.22(q,J=17.6Hz,1H),3.84(d,J=7.0Hz,1H),3.96-4.01(m,1H),4.21(d,J=8.4Hz,1H),4.31(d,J=8.2Hz,1H),4.44(dd,J1=12.4Hz,J 2=6.8Hz,1H),4.58(m,1H),4.98(d,J=8.6Hz,1H),5.09(d,2H),5.44(d,J=2.8Hz,1H),5.71(d,J=6.8Hz,1H),6.01(dd,J1=12.4Hz,J2=3Hz,1H),6.03(d,J=8.8Hz,1H),6.24(t,J=9.1Hz,1H),6.30(s,1H),6.4(t,J=5.6Hz,1H),7.17-7.6(m,21H),8.14(d,J=6.8Hz,2H).
10.2N-(紫杉醇-2′-氧羰基丁酰)-L-甘氨酸(Ia-10)的制备
将IIIa-10 340mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-10的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1023.9(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:0.99(3H,s,17-H),1.04(3H,s,16-H),1.498(3H,s,19-H),1.63(1H,t,J=8.8Hz),1.78(6H,m),2.10(3H,s,10-OAc),2.16(2H,t,J=7.2Hz,-CH2COOR),2.257(3H,s,4-OAc),2.43(6H,t,J=7.2Hz,-CH2COOR),3.60(3H,m,6″-H,3-H),3.99-4.03(2H,q,J=12.4Hz,20-H),4.12(1H,m,7-H),),4.67(1H,s,5-H),4.92(2H,t,J=6.8Hz),5.35(1H,d,J=8.4Hz,2’-H),5.41(1H,d,J=6.8Hz,2-H),5.53(1H,q,J=5.5Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.30(1H,s,10-H),7.18-8.00(16H,m),9.30(1H,d,J=8.8Hz,NH).
实施例11N-(紫杉醇-2′-氧羰基丁酰)-L-异亮氨酸(Ia-11)的制备
11.1N-(紫杉醇-2′-氧羰基丁酰基)-L-异亮氨酸苄酯(IIIa-11)的制备
25ml烧瓶中加入IIa-2 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Ile(OBzl)-OBzl·TosOH 179mg(MW=393.5,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-11白色粉末状固体351mg。产率73.9%。MS(ESI)m/z:1171.7(M+H+),1H-NMR(400M Hz,DMSO-d6)δppm:0.777-0.803(6H,m,3″′-H3,2″′-CH3),1.0(3H,s,17-H),1.03(3H,s,16-H),1.5(3H,s,19-H),1.78(4H,brs),2.10(3H,s,10-OAc),2.20(6H,m),2.246(3H,s,4-OAc),2.40(2H,t,J=7.2Hz,4″-CH2COO),3.60(1H,d,J=7.2,3-H),4.01(2H,q,J=12.4Hz,20-H2),4.12-4.23(2H,m,7″-H,7-H),4.62(1H,s,5-H),4.91(2H,d,J=9.8),5.12(2H,d,J=8.4Hz,-COOCH2Ph),5.34(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H),5.51(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-8.03(21H,m),9.21(1H,d,J=8.4Hz,NH).
11.2N-(紫杉醇-2′-氧羰基丁酰)-L-异亮氨酸(Ia-11)的制备
将IIIa-11 351mg用甲醇20ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,真空干燥,得Ia-11的白色粉末状固体324mg,产率100%。MS(ESI)m/z:1080.0(M-H+),1H-NMR(400M Hz,DMSO-d6)δppm:0.788-1.3(14H,m,3″′-H3,2″′-CH3,17-H3,16-H3,2″′-H,1″′-H),1.497-1.785(11H,m),2.10(3H,s,10-OAc),2.16(2H,t,J=7.2,2″′-H),2.20(2H,m,2″-H),2.20(2H,t,J=7.2Hz,2″-CH2COO),2.246(3H,s,4-OAc),2.416(2H,t,J=7.2Hz,4″-CH2COO),3.60(3H,m,7″-H,3-H),4.01(2H,q,J=12.4Hz,20-H2),4.12(2H,m,7″-H,7-H),4.66(1H,s,5-H),4.94(2H,d,J=9.8),5.34(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H)5.51(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-8.03(16H,m),9.278(1H,d,J=8.4Hz,NH).
实施例12N-(紫杉醇-2′-氧羰基丁酰)-L-异亮氨酸(Ia-12)的制备
12.1N-(紫杉醇-2′-氧羰基丁酰)-L-亮氨酸苄酯(IIIa-12)的制备
25ml烧瓶中加入IIa-2 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Leu(OBzl)-OBzl·TosOH 179mg(MW=393.5,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-12的白色粉末状固体332mg。产率71.2%。MS(ESI)m/z:1171.7(M+H+),1H-NMR(400MHz,DMSO-d6)δppm:0.80(3H,d,J=6.4Hz,2″′-CH3),0.840(3H,t,J=6.0Hz,2″′-CH3),0.99(3H,s,17-H),1.03(3H,s,16-H),1.46(4H,m),1.74(3H,d,J=6.4Hz,2″-H),1.78(3H,s,18-H),2.10(3H,s,10-OAc),2.16(2H,t,J=7.2,2″′-H),2.26(3H,s,4-OAc),2.43(2H,t,J=7.2Hz,4″-CH2COO),3.58(1H,d,J=7.2Hz,3-H),4.00-4.13(4H,m,20-H,7″-H,7-H),4.67(1H,s,5-H),4.91(2H,d,J=9.8Hz),5.06(2H,s,-COOCH2Ph),5.34(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H)5.50(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(21H,m),9.31(1H,d,J=8.4Hz,NH).
12.2N-(紫杉醇-2′-氧羰基丁酰)-L-亮氨酸(Ia-12)的制备
将IIIa-12351mg用甲醇30ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-12的白色粉末状固体306mg,产率100%。MS(ESI)m/z:1080.0(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:0.79(15H,m,2″′-CH3,2″′-CH3,17-H,16-H),1.46(1H,m,1″′-H),1.458(4H,brs,1.74(3H,d,J=6.4Hz,2″-H),1.78(3H,s,18-H),2.10(3H,s,10-OAc),2.16(2H,t,J=7.2,2″′-H),2.20(2H,m,2″-H),2.25(3H,s,4-OAc),2.42(2H,t,J=7.2Hz,4″-CH2COO),3.168(1H,s),3.60(3H,m,7″-H,3-H),4.05(2H,q,J=12.4Hz,20-H2),4.12(2H,m,7″-H,7-H),4.67(1H,s,5-H),4.91(2H,d,J=9.8Hz),5.35(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H),5.52(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(16H,m),9.31(1H,d,J=8.4Hz,N H).
实施例13N-(紫杉醇-2′-氧羰基丁酰)-L-缬氨酸(Ia-13)的制备
13.1N-(紫杉醇-2′-氧羰基丁酰)-L-缬氨酸苄酯(IIIa-13)的制备
25ml烧瓶中加入IIa-2400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Val(OBzl)-OBzl·TosOH 179mg(MW=393.5,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIa-13的白色粉末状固体344mg。产率72%。MS(ESI)m/z:1157.9(M+H+),1H-NMR(400M Hz,DMSO-d6)δppm:0.83(3H,d,J=2.0Hz,1″′-CH3),0.84(3H,d,J=2.0Hz,1″′-CH3),0.99(3H,s,17-H),1.03(3H,s,16-H),1.04(2H,s),1.498-1.78(11H,m,19-H,3″-H,18-H),2.02(1H,m,1″′-H),2.10(3H,s,10-OAc),2.20-2.41(8H,m,2″-H,4-OAc,4″-CH2COO),358(1H,d,J=7.6Hz,3-H),3.95-4.01(2H,q,J=12.4Hz,20-H),4.06-4.19(2H,m,7″-H,7-H),4.62(1H,s,5-H),4.9(2H,d,J=6.8Hz),5.12(2H,s,-COOCH2Ph),5.38(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H),5.53(1H,t,J=8.4Hz,3’-H),5.82(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.16-8.12(16H,m),9.21(1H,d,J=8.4Hz,NH).
13.2N-(紫杉醇-2′-氧羰基丁酰)-L-缬氨酸(Ia-13)的制备
将N IIIa-13334mg用甲醇30ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-13白色粉末状固体317mg,产率100%。MS(ESI)m/z:1065.9(M-H+),1H-NMR(400M Hz,DMSO-d6)δppm:0.81(3H,d,J=2.0Hz,1″′-CH3),0.82(3H,d,J=2.0Hz,1″′-CH3),0.99(3H,s,17-H),1.03(3H,s,16-H),1.498(3H,s,19-H),1.75(2H,m,3″-H),1.78(3H,s,18-H),2.02(1H,m,1″′-H),2.10(3H,s,10-OAc),2.20(2H,m,2″-H),2.26(3H,s,4-OAc),2.44(2H,t,J=7.2Hz,4″-CH2COO),3.60(3H,m,7″-H,3-H),3.95-4.01(2H,q,J=12.4Hz,20-H),4.06-4.12(2H,m,7″-H,7-H),4.66(1H,s,5-H),4.92(2H,d,J=9.8Hz),5.34(1H,d,J=7.2Hz,2’-H),5.40(1H,d,J=7.2Hz,2-H)5.50(1H,t,J=8.4Hz,3’-H),5.81(1H,t,J=8.4Hz,13-H),6.3(1H,s,10-H),7.18-7.99(16H,m),9.28(1H,d,J=8.4Hz,NH).
实施例14N-(多烯紫杉醇-2′-氧羰基丁酰)-D-天冬氨酸(Ib-1)的制备
14.1N-(紫杉醇-2′-氧羰基丁酰)-D-天冬氨酸苄酯(IIIa-14)的制备
25ml烧瓶中加入IIa-2 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq),溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-D-Asp(OBzl)-OBzl·TosOH 200mg(MW=485.6,1.1eq)及N-甲基吗啡啉(NMM)64μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出得的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶异丙醇=35∶1)。得IIIa-14的白色粉末状固体350mg。产率68%。MS(ESI)m/z:1263(M+H+),1H-NMR(400M Hz,DMSO-d6)δppm:1.13-1.25(m,6H),1.69(s,3H),1.90(brs.4H),2.1-2.22(m,5H),2.52(brs,4H),2.74-2.97(m,3H),3.82(d,J=7.0Hz,1H),3.84(m,1H),4.22(d,J=8.4Hz,1H),4.30(d,J=8.2Hz,1H),4.46(dd,J1=12.4Hz,J2=6.8Hz,1H),4.71(m,1H),4.98(d,J=8.6Hz,1H),5.05-5.12(ss,4H),5.54(d,J=2.8Hz,1H),5.69(d,J=6.8Hz,1H),6.01(dd,J1=12.4Hz,J2=3Hz,1H),6.29(t,J=9.1Hz,1H),6.30(s,1H),6.78(t,J=5.6Hz,1H),7.22-7.73(m,26H),8.15(d,J=6.8Hz,2H).
14.2N-(紫杉醇-2′-氧羰基丁酰)-D-天冬氨酸(Ia-14)的制备
将IIIa-14 350mg用甲醇30ml溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,用真空泵蒸干滤液,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),真空干燥,得Ia-14白色粉末状固体305mg,产率100%。MS(ESI)m/z:1081.7(M-H+),1H-NMR(400MHz,DMSO-d6)δppm:0.99(3H,s,17-H),1.04(3H,s,16-H),1.50-1.85(12H,m,19-H,18-H),2.10-2.15(5H,m,10-OAc),2.26(3H,s,4-OAc),2.42-2.60(6H,m,-CH2COOR),3.59(1H,d,J=7.2Hz,3-H),3.99-4.01(2H,q,J=12.4Hz,20-H),4.12(1H,m,7-H),4.4(1H,q,J=4.4Hz,6″-H),4.66(1H,s,5-H),4.93(2H,t,J=6.8Hz),5.34(1H,d,J=8.4Hz,2’-H),5.41(1H,d,J=6.8Hz,2-H),5.52(1H,q,J=5.5Hz,3’-H),5.81(1H,t,J=9.0Hz,13-H),6.30(1H,s,10-H),7.18-7.99(16H,m),9.26(1H,d,J=8.8Hz,N H),13.423(br-s,-COOH).
实施例15N-(多西紫杉醇-2′-氧羰基丙酰)-L-天冬氨酸(Ib-1)的制备
15.1多西紫杉醇-2′-琥珀酸单酯(IIb-1)的制备
将1200mg(1.404mmol)多西紫杉醇,168.8mg(1.688mmol)琥珀酸酐,36mg(0.28mmol)DMAP溶于40ml无水吡啶中。室温搅拌3小时。旋干溶剂,残留物用少量二氯甲烷溶解,柱层析(二氯甲烷∶甲醇=10∶1)得IIb-1的白色粉末状固体1330mg,产率99.3%。MS(ESI)m/z:908.9(M+H+),1H-NMR(CDCl3):δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);6.38(s,1H);5.80(t,1H);5.57(t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);2.36-2.60(m,10H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
15.2N-(多西紫杉醇-2′-氧羰基丙酰)-L-天冬氨酸苄酯(IIIb-1)的制备
25ml烧瓶中加入IIb-1 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Asp(OBzl)-OBzl·TosOH 225mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIb-1的白色粉末状固体365mg。产率71.3%MS(ESI)m/z:1203.3(M+H+),1H-NMR(CDCl3):δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);7.18-7.99(10H,m),6.38(s,1H);5.80(t,1H);5.57(t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,4H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
15.3N-(多西紫杉醇-2′-氧羰基丙酰)-L-天冬氨酸(Ib-1)的制备
将IIIb-1 365mg加入30ml甲醇溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除滤剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得Ib-1的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1023.7(M-H+),1H-NMR(400M Hz DMSO-d6)δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);6.38s,1H);5.80t,1H);5.57t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,4H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
实施例16N-(多西紫杉醇-2′-氧羰基丙酰)-L-谷氨酸(Ib-2)的制备
16.1N-(多西紫杉醇-2′-氧羰基丙酰)-L-谷氨酸苄酯(IIIb-2)的制备
25ml烧瓶中加入IIb-1 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Glu(OBzl)-OBzl·TosOH 225mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIb-2的白色粉末状固体365mg。产率71.3%MS(ESI)m/z:1217.3(M+H+),1H-NMR(CDCl3):δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);7.18-7.99(10H,m),6.38(s,1H);5.80(t,1H);5.57(t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,6H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
16.2N-(多西紫杉醇-2′-氧羰基丙酰)-L-谷氨酸(Ib-2)的制备
将IIIb-2365mg加入30ml甲醇溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除滤剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得Ib-2的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1037.7(M-H+),1H-NMR(400M Hz DMSO-d6)δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);6.38s,1H);5.80t,1H);5.57t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,4H),2.41(s,3H),2.30(m,3H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
实施例17N-(多西紫杉醇-2′-氧羰基丁酰)-L-天冬氨酸(Ib-3)的制备
17.1多西紫杉醇-2′-戊二酸单酯(Ib-2)的制备
将1200mg(1.404mmol)多西紫杉醇,168.8mg(1.688mmol)戊二酸酐,36mg(0.28mmol)DMAP溶于40ml无水吡啶中。室温搅拌3小时。旋干溶剂,残留物用少量二氯甲烷溶解,柱层析(二氯甲烷∶甲醇=10∶1)得Ib-2的白色粉末状固体1330mg,产率99.3%。MS(ESI)m/z:922.4(M+H+),1H-NMR(CDCl3):δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);6.38(s,1H);5.80(t,1H);5.57(t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);2.36-2.60(m,12H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
17.2N-(多西紫杉醇-2′-氧羰基丁酰)-L-天冬氨酸苄酯(IIIb-3)的制备25ml IIb-2 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Asp(OBzl)-OBzl·TosOH 225mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIb-3的白色粉末状固体365mg。产率71.3%MS(ESI)m/z:1217.6(M+H+),1H-NMR(CDCl3):δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);7.18-7.99(10H,m),6.38(s,1H);5.80(t,1H);5.57(t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,6H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
17.3N-(多西紫杉醇-2′-氧羰基丁酰)-L-天冬氨酸(Ib-3)的制备
将IIIb-3 365mg加入30ml甲醇溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除滤剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得I b-3的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1037.9(M-H+),1H-NMR(400M Hz DMSO-d6)δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);6.38s,1H);5.80t,1H);5.57t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,6H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
实施例18N-(多西紫杉醇-2′-氧羰基丁酰)-L-谷氨酸(Ib-4)的制备
18.1N-(多西紫杉醇-2′-氧羰基丁酰)-L-谷氨酸苄酯(IIIb-4)的制备
25ml烧瓶中加入IIb-2 400mg,加入无水二氯甲烷20ml使溶,冰浴,电磁搅拌。再加入HOBT 68mg(1.2eq)溶于5ml DMF的溶液。冷却到0℃后,加入DCC 86mg(1.2eq)溶于无水二氯甲烷的溶液。冰浴反应2小时。再加入H-Glu(OBzl)-OBzl·TosOH 225mg(1.1eq)及N-甲基吗啡啉(NMM)66μl溶于10ml无水二氯甲烷的溶液,撤冰浴,室温反应8小时。反应完成。滤除析出的DCU,减压蒸除溶剂,加EtAc 50ml溶解,再次滤除析出的DCU,EtAc层分别用5%的碳酸氢钠洗30ml×3次,0.5N HCl洗25ml×2次,5%的碳酸氢钠洗30ml×3次,饱和NaCl洗×3次,加无水硫酸镁干燥4-6小时。然后滤去干燥剂,蒸除溶剂。柱层析(二氯甲烷∶甲醇=10∶1)。得IIIb-4的白色粉末状固体365mg。产率71.3%MS(ESI)m/z:1231.7(M+H+),1H-NMR(CDCl3):δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);7.18-7.99(10H,m),6.38(s,1H);5.80(t,1H);5.57(t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,8H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
18.2N-(多西紫杉醇-2′-氧羰基丁酰)-L-谷氨酸(Ib-4)的制备
将IIIb-4 365mg加入30ml甲醇溶解,加入10%Pd/C 30mg,催化氢化5小时,反应完成。用微孔滤过膜(有机相)滤出Pd/C,蒸除滤剂,柱层析(甲醇∶二氯甲烷∶甲酸=10∶1∶0.1),得Ib-4的白色粉末状固体310mg,产率100%。MS(ESI)m/z:1051.9(M-H+),1H-NMR(400M Hz DMSO-d6)δppm:9.25(d,1H);8.48(brs,2H);8.05-7.32(m,10H);6.38s,1H);5.80t,1H);5.57t,1H);5.42(d,1H);5.36(d,1H);4.94(dd,1H);4.82(brs,1H);4.61(s,1H);4.14(dd,1H);3.95-4.06(m,3H);3.57(d,1H);3.22(q,J=17.6Hz,1H),2.74-2.83(m,2H),2.51(m,8H),2.41(s,3H),2.30(m,1H),2.22(s,3H),2.11(m,1H),1.86(s,3H);1.80(dd,1H);1.64(t,1H);1.52(s,3H);1.40(s,9H);1.06(s,3H);1.02(s,3H)。
实施例19目标化合物体外释放度评价
19.1仪器与试剂
高效液相色谱仪岛津LC-15C;紫外检测器 岛津SPD-15C;色谱柱 不锈钢反相C18柱(250×4.6mm);流动相(低压洗脱):三氟乙酸-超纯水-乙腈(1∶500∶500);检测波长:227nm;pH=7.4缓冲液自制;人体血浆;进样量20μl。
通过紫外吸收光谱扫描,测定目标化合物最大吸收波长为227nm,因此选择波长227nm为HPLC检测波长。
19.2目标化合物在PBS缓冲溶液(PH值=7.4)中的释放度试验
将一定质量的偶合物置于少量水中,用1N碳酸氢钠调pH至7左右,加缓冲液至2ml(药物浓度约为0.2mmol/l),按照体积0.2ml分成10等份,于37℃孵育,在不同的时间(30min,1h,2h,4h,12h,24h等)取样,HPLC检测,用水和乙腈稀释到0.5ml,用4ml甲基叔丁基醚萃取两次,涡流30s混匀,离心(16000rms约10min)取上层清液,合并有机相,氮气吹干。所得固体用100μl流动相溶解,HPLC监测,观察偶合物与Paclitaxel处峰面积,以游离药物与偶合物峰面积之和为100%,计算药物释放度。
19.3目标化合物在人血浆中释放度实验
将样品与碳酸氢钠以摩尔比1∶1.2溶于0.1N碳酸氢钠溶液中,加入100μl生理盐水稀释,然后加入到1ml血浆中,配成0.2mmol/l的溶液。涡流混合,隔定量时间取样50μl,加入100μl冷甲醇稀释,沉淀蛋白,涡流混合30s,离心(13000rms约15min),取上清液,HPLC进样监测。以内标的峰面积为100%计算偶合物释放的药物度。
表1目标化合物在缓冲液和人血浆中48小时内药物释放度
(释放的游离药物在目标化合物中的百分比,%)
编号 | PBS 7.4(48h) | 人血浆(48h) |
Ia-1 | 0.05 | 54.7 |
Ia-2 | 0.2 | 37 |
Ia-3 | 1.3 | 43 |
Ia-4 | 1.2 | 35 |
Ia-5 | 2.8 | 38.2 |
Ia-6 | 5.9 | 28.9 |
Ia-7 | 0 | 34.9 |
Ia-8 | 0.1 | - |
Ia-9 | 1.8 | 29 |
Ia-10 | 0.8 | - |
Ia-11 | 1.3 | 25.3 |
Ia-12 | 4.0 | - |
Ia-13 | 0.3 | 35.2 |
实施例20目标化合物体外细胞毒性活性评价
用SRB(磺基罗丹明B)染色法测定细胞活力检测目标化合物体外细胞毒性。
将人非小细胞肺癌A549细胞株以5000个/孔接种至96孔板,过夜孵育后给予不同浓度受试化合物,继续培养72h后弃去培养基,每孔轻柔加入100μL冰冷的10%三氯醋酸,置4℃固定1h,用蒸馏水洗涤4次,空气干燥。加入70μL 0.4%(w/v)磺基罗丹明B(于1%乙酸中),室温染色30min,弃染色液用1%乙酸冲洗4次,空气彻底干燥。每孔加入10mM Tris碱溶液(pH10.5)100μL完全水解,用酶标仪测定560nM波长吸光度值。以不加药对照孔吸光度值为100%,计算加药孔相对百分数即为细胞活力百分数。
表2目标化合物细胞毒活性IC50值
编号 | IC50(nM) | 编号 | IC50(nM) |
Ia-1 | 4.9 | Ia-9 | 4.3 |
Ia-2 | 7.6 | Ia-10 | 3.2 |
Ia-3 | 9.0 | Ia-11 | 2.8 |
Ia-4 | 3.3 | Ia-12 | 5.9 |
Ia-5 | 8.6 | Ib-1 | 2.5 |
Ia-6 | 5.9 | Ib-2 | 4.1 |
Ia-7 | 7.5 | Ib-3 | 4.7 |
Ia-8 | 6.2 | Ib-4 | 3.2 |
紫杉醇 | 3.3 |
Claims (6)
4.权利要求1-3所述的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物的非毒性药学上可接受的盐为紫杉醇及多西紫杉醇的水溶性衍生物的羧基与无机金属离子的钠盐、钾盐或与有机胺的盐。
5.权利要求1-3所述的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐作为活性成分的药物组合物。
6.权利要求1-3所述的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐,以及含有式I所示的紫杉醇类抗肿瘤药物紫杉醇及多西紫杉醇的水溶性衍生物及其非毒性药学上可接受的盐作为活性成分的药物组合物作为抗肿瘤药物的用途。
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CN103739576A (zh) * | 2014-01-24 | 2014-04-23 | 北京普瑞博思投资有限公司 | 一种新型抗病毒穿心莲内酯衍生物及其制备方法和应用 |
CN104370862A (zh) * | 2013-08-13 | 2015-02-25 | 中国人民解放军军事医学科学院毒物药物研究所 | 水溶性抗肿瘤化合物 |
CN109422759A (zh) * | 2017-08-22 | 2019-03-05 | 复旦大学 | 小分子修饰的紫杉烷类水溶性前药及其药用用途 |
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CN101028259A (zh) * | 1996-03-12 | 2007-09-05 | Pg-Txl有限公司 | 水溶性紫杉醇产品 |
US5981564A (en) * | 1998-07-01 | 1999-11-09 | Universite Laval | Water-soluble derivatives of paclitaxel, method for producing same and uses thereof |
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CN104370862A (zh) * | 2013-08-13 | 2015-02-25 | 中国人民解放军军事医学科学院毒物药物研究所 | 水溶性抗肿瘤化合物 |
CN104370862B (zh) * | 2013-08-13 | 2019-04-23 | 中国人民解放军军事医学科学院毒物药物研究所 | 水溶性抗肿瘤化合物 |
CN103739576A (zh) * | 2014-01-24 | 2014-04-23 | 北京普瑞博思投资有限公司 | 一种新型抗病毒穿心莲内酯衍生物及其制备方法和应用 |
CN109422759A (zh) * | 2017-08-22 | 2019-03-05 | 复旦大学 | 小分子修饰的紫杉烷类水溶性前药及其药用用途 |
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