叶酸介导的牛血清白蛋白氟尿嘧啶前体药物的合成及靶向作用的研究
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摘要
目的:合成叶酸介导的牛血清白蛋白5-氟尿嘧啶偶联物(F-BSA-5-FU),并进行各步反应纯化、分离及合成条件的优化;制备叶酸介导的牛血清白蛋白5-氟尿嘧啶偶联物纳米粒(F-BSANP-5-FU)及牛血清白蛋白5-氟尿嘧啶偶联物纳米粒(BSANP-5-FU),通过制备包裹钙黄绿素纳米粒对偶联物体内肿瘤靶向性进行评价,并对偶联物纳米粒药效学进行评价。
方法:运用正交实验设计叶酸偶联牛血清白蛋白的合成实验,以叶酸用量、EDC与叶酸摩尔比、反应时间三因素为考察对象,以叶酸与牛血清白蛋白的偶联比值为评价指标,从中筛选出最优的合成条件。以产率为指标进行5-氟尿嘧啶-1-基乙酸(5-FUA)及其活性酯的合成,运用正交设计5-FUA活性酯的合成实验,筛选最佳合成条件,计算产率,测定产物熔点。通过单因素考察5-FUA活性酯与BSA、F-BSA的偶联条件,优化最佳合成条件。分别运用紫外、红外、差热分析、MALDI-TOF-MS监测上述合成产物,同时通过化学分析法及MALDI-TOF-MS计算F-BSA-5-FU、BSA-5-FU中药物密度,即前体药物中偶联5-FU的个数。采用去溶剂-化学交联法制备F-BSANP-5-FU、BSANP-5-FU及包裹有钙黄绿素的纳米粒,以药物浓度、溶液pH、交联剂的用量、交联时间为考察对象,以纳米粒平均粒径及多分散系数为考察指标,优化纳米粒的制备条件,通过透射电镜、激光粒度分布仪等对纳米粒进行表征;以接种7～10d的肝癌(H22 )实体瘤小鼠为模型,腹腔注射包裹有钙黄绿素的纳米粒,采用荧光分光光度法评价纳米粒的体内肿瘤靶向性情况;通过腹腔给药评价F-BSANP-5-FU、BSANP-5-FU抑制肿瘤情况,并计算抑瘤率、脾指数、胸腺指数。将原始数据输入微机,利用SPSS11.5进行统计分析。抑瘤率(%)=(对照组平均瘤重-给药组平均瘤重)/对照组平均瘤重×100% ;脾和胸腺指数计算方法如下:脾或胸腺指数=脾或胸腺重量(mg)/鼠体重(g),对各组的肿瘤进行病理切片,观察肿瘤抑制情况。
结果:运用最优化的合成条件合成了偶联比约为4的叶酸牛血清白蛋白,即一个牛血清白蛋白偶联4个叶酸;5-FU mp:283℃～284℃,5-FUA mp:276℃～278℃,产率为81.37%;5-FUA活性酯mp:255℃～257℃,产率为83.98%;紫外、红外、差热分析、MALDI-TOF-MS结果显示已成功合成上述化合物;通过化学分析法及MALDI-TOF-MS计算F-BSA-5-FU、BSA-5-FU中偶联5-FU的个数分别为11、13;成功制备粒径均小于200nm的纳米粒,所得纳米粒粒径均一,形态完整;体内肿瘤靶向性实验显示:腹腔注射同等剂量不同组别的含钙黄绿素药物在体内的蓄积不同(P<0.05);包裹有钙黄绿素的BSANP-5-FU组(A组)在肿瘤组织中的钙黄绿素量在2.0h达到峰值,而后下降,包裹有钙黄绿素的F-BSANP-5-FU组(B组)在肿瘤组织中的钙黄绿素浓度在0.5h就达到峰值,而后急剧下降,肝、肾组织中的钙黄绿素量低于肿瘤组织,却高于心、脾、肺组织,B组峰值显著高于A组;动物水平的药效学实验显示:腹腔注射F-BSANP-5-FU高剂量组的荷瘤小鼠肿瘤生长最为缓慢,抑瘤率最高,达到47.19%,F-BSANP-5-FU的抑瘤率显著高于BSANP-5-FU,在加入1 mmol·L-1外源性叶酸后,F-BSANP-5-FU的抑瘤率明显降低。5-FU阳性对照组的抑瘤率高于BSANP-5-FU组。
结论:采用温和条件下合成的F-BSA-5-FU,其纳米粒具有肿瘤靶向性,该靶向性是通过叶酸介导的,并且对肿瘤有很好的抑制作用,是一种有潜力的抗肿瘤的前体药物,可靶向于高表达叶酸受体的肿瘤。
Objective: The prodrug folate-mediated bovine serum albumin -5-fluorouracil (F-BSA-5-FU)was synthesized,then purify、separate and optimize the synthesis conditions of it;Folate-mediated bovine serum albumin 5 - fluorouracil nanoparticles (F-BSANP-5-FU) and bovine serum albumin 5 - fluorouracil nanoparticles (BSANP-5-FU) were prepared; Tumor targeting in vivo through the preparation of loaded calcein nanoparticles and the pharmacodynamics of the conjugates nanoparticles were evaluated respectively.
Methods: The synthesis of folate- bovine serum albumin was designed using orthogonal experiment;the effect factors such as the amount of folate、the molar ratio of EDC and folate、the reaction time were investigated respectively.Using the coupled rate of folate and bovine serum albumin as the evaluation of indicators, selected the optimal synthesis conditions. Using the yield as the evaluation of indicators, the 5-fluorouracil-1-acetic acid (5-FUA) and its active ester were synthesized, The synthesis of 5-FUA active ester was designed using orthogonal experiment, selected the optimal synthesis conditions、calculate yield、determine melting point of the product. The coupling of 5-FUA active ester with BSA and F-BSA were investigated,the best synthesis conditions was optimizated through the single-factor analysis.The above-mentioned products were monitored by ultraviolet、infrared、differential thermal analysis、MALDI-TOF-MS.Simultaneously the content of 5-FU in F-BSA-5-FU and BSA-5-FU were calculated by chemical methods and MALDI-TOF MS. F-BSANP-5-FU、BSANP-5-FU and loaded calcein nanoparticles were prepared by desolvation-chemical coupling method, the effect factors such as the concentration of drug、the pH of the solution、the amount of coupling agent、the coupling time were investigated. Using the average diameter and polyindex as the evaluation of indicators, selected the optimal preparation conditions, and the nanoparticles were characterized. Using tumor-bearing mice which was inoculated 7 ~ 10d for liver cancer (H22) solid tumor as animal model,the tumor targeting in vivo was determined by spectrophotofluorometer through the loaded calcein nanoparticles.The tumor inhibition information of F-BSANP-5-FU and BSANP-5-FU were evaluated by intraperitoneal injection, Original data was input to computer and was analyzed by the statistical software SPSS11.5.Inhibition rate (%)=( average tumor weight in contrast group - average tumor weight in dosage group) / average tumor weight in contrast group×100%; spleen and thymus index calculated as follows: spleen or thymus index = spleen or thymus weight (mg ) / mouse weight (g),the tumor of each group was made pathological section and the information of tumor suppression was observed .
Results: The coupling ratio of about 4 of folate-bovine serum albumin was synthesized with the most optimized synthesis conditions,that is a bovine serum albumin was coupled with 4 folates; 5-FU melting point was 283℃～284℃,5-FUA melting point was 276℃～278℃, yield was 81.37%;5-FUA active ester melting point was 255℃～257℃, yield was 83.98%;The results of ultraviolet、infrared、differential thermal analysis、MALDI-TOF MS showed that compounds were synthesized successfully;The nanoparticles whose particle size was less than 200nm were prepared successfully, and the size was uniform, the shape was integrated;The tumor targeting in vivo experiments showed that the equal doses of different groups of calcein were given by intraperitoneal injection,the accumulation of the content of calcein were different in vivo (P <0.05);The content of loaded calcein the BSANP-5-FU group (A group) reached its peak at 2.0h, and then declined in tumor tissues, The content of loaded calcein of Folate-BSANP-5-FU group (B group) reached its peak at 0.5h, and then declined sharply in tumor tissues . The content of calcein of liver and kidney tissues were less than the tumor tissue, but higher than the heart、spleen、lung tissue.The peak value of B group was significantly higher than A Group; The level of animal pharmacodynamic experiments showed that the tumor growth was the slowest in the tumor-bearing mice by intraperitoneal injection of F-BSANP-5-FU high-dose group, the highest inhibition rate reached 47.19%, the inhibition rate of F-BSANP-5-FU was significantly higher than BSANP-5-FU, after adding 1 mmol ? L-1 exogenous folate, the inhibition rate of F-BSANP-5-FU was declined significantly. The inhibition rate 5-FU positive control group was higher than BSANP-5-FU.
Conclusions: F-BSA-5-FU was synthesized under mild conditions, and its nanoparticles had tumor targeting, the targeting was likely to pass the folate-mediated, F-BSA-5-FU had good the inhibition of tumor,and its was a potential anti-tumor drugs that can be targeted at tumor with the high expression of folate receptor.
方法:运用正交实验设计叶酸偶联牛血清白蛋白的合成实验,以叶酸用量、EDC与叶酸摩尔比、反应时间三因素为考察对象,以叶酸与牛血清白蛋白的偶联比值为评价指标,从中筛选出最优的合成条件。以产率为指标进行5-氟尿嘧啶-1-基乙酸(5-FUA)及其活性酯的合成,运用正交设计5-FUA活性酯的合成实验,筛选最佳合成条件,计算产率,测定产物熔点。通过单因素考察5-FUA活性酯与BSA、F-BSA的偶联条件,优化最佳合成条件。分别运用紫外、红外、差热分析、MALDI-TOF-MS监测上述合成产物,同时通过化学分析法及MALDI-TOF-MS计算F-BSA-5-FU、BSA-5-FU中药物密度,即前体药物中偶联5-FU的个数。采用去溶剂-化学交联法制备F-BSANP-5-FU、BSANP-5-FU及包裹有钙黄绿素的纳米粒,以药物浓度、溶液pH、交联剂的用量、交联时间为考察对象,以纳米粒平均粒径及多分散系数为考察指标,优化纳米粒的制备条件,通过透射电镜、激光粒度分布仪等对纳米粒进行表征;以接种7～10d的肝癌(H22 )实体瘤小鼠为模型,腹腔注射包裹有钙黄绿素的纳米粒,采用荧光分光光度法评价纳米粒的体内肿瘤靶向性情况;通过腹腔给药评价F-BSANP-5-FU、BSANP-5-FU抑制肿瘤情况,并计算抑瘤率、脾指数、胸腺指数。将原始数据输入微机,利用SPSS11.5进行统计分析。抑瘤率(%)=(对照组平均瘤重-给药组平均瘤重)/对照组平均瘤重×100% ;脾和胸腺指数计算方法如下:脾或胸腺指数=脾或胸腺重量(mg)/鼠体重(g),对各组的肿瘤进行病理切片,观察肿瘤抑制情况。
结果:运用最优化的合成条件合成了偶联比约为4的叶酸牛血清白蛋白,即一个牛血清白蛋白偶联4个叶酸;5-FU mp:283℃～284℃,5-FUA mp:276℃～278℃,产率为81.37%;5-FUA活性酯mp:255℃～257℃,产率为83.98%;紫外、红外、差热分析、MALDI-TOF-MS结果显示已成功合成上述化合物;通过化学分析法及MALDI-TOF-MS计算F-BSA-5-FU、BSA-5-FU中偶联5-FU的个数分别为11、13;成功制备粒径均小于200nm的纳米粒,所得纳米粒粒径均一,形态完整;体内肿瘤靶向性实验显示:腹腔注射同等剂量不同组别的含钙黄绿素药物在体内的蓄积不同(P<0.05);包裹有钙黄绿素的BSANP-5-FU组(A组)在肿瘤组织中的钙黄绿素量在2.0h达到峰值,而后下降,包裹有钙黄绿素的F-BSANP-5-FU组(B组)在肿瘤组织中的钙黄绿素浓度在0.5h就达到峰值,而后急剧下降,肝、肾组织中的钙黄绿素量低于肿瘤组织,却高于心、脾、肺组织,B组峰值显著高于A组;动物水平的药效学实验显示:腹腔注射F-BSANP-5-FU高剂量组的荷瘤小鼠肿瘤生长最为缓慢,抑瘤率最高,达到47.19%,F-BSANP-5-FU的抑瘤率显著高于BSANP-5-FU,在加入1 mmol·L-1外源性叶酸后,F-BSANP-5-FU的抑瘤率明显降低。5-FU阳性对照组的抑瘤率高于BSANP-5-FU组。
结论:采用温和条件下合成的F-BSA-5-FU,其纳米粒具有肿瘤靶向性,该靶向性是通过叶酸介导的,并且对肿瘤有很好的抑制作用,是一种有潜力的抗肿瘤的前体药物,可靶向于高表达叶酸受体的肿瘤。
Objective: The prodrug folate-mediated bovine serum albumin -5-fluorouracil (F-BSA-5-FU)was synthesized,then purify、separate and optimize the synthesis conditions of it;Folate-mediated bovine serum albumin 5 - fluorouracil nanoparticles (F-BSANP-5-FU) and bovine serum albumin 5 - fluorouracil nanoparticles (BSANP-5-FU) were prepared; Tumor targeting in vivo through the preparation of loaded calcein nanoparticles and the pharmacodynamics of the conjugates nanoparticles were evaluated respectively.
Methods: The synthesis of folate- bovine serum albumin was designed using orthogonal experiment;the effect factors such as the amount of folate、the molar ratio of EDC and folate、the reaction time were investigated respectively.Using the coupled rate of folate and bovine serum albumin as the evaluation of indicators, selected the optimal synthesis conditions. Using the yield as the evaluation of indicators, the 5-fluorouracil-1-acetic acid (5-FUA) and its active ester were synthesized, The synthesis of 5-FUA active ester was designed using orthogonal experiment, selected the optimal synthesis conditions、calculate yield、determine melting point of the product. The coupling of 5-FUA active ester with BSA and F-BSA were investigated,the best synthesis conditions was optimizated through the single-factor analysis.The above-mentioned products were monitored by ultraviolet、infrared、differential thermal analysis、MALDI-TOF-MS.Simultaneously the content of 5-FU in F-BSA-5-FU and BSA-5-FU were calculated by chemical methods and MALDI-TOF MS. F-BSANP-5-FU、BSANP-5-FU and loaded calcein nanoparticles were prepared by desolvation-chemical coupling method, the effect factors such as the concentration of drug、the pH of the solution、the amount of coupling agent、the coupling time were investigated. Using the average diameter and polyindex as the evaluation of indicators, selected the optimal preparation conditions, and the nanoparticles were characterized. Using tumor-bearing mice which was inoculated 7 ~ 10d for liver cancer (H22) solid tumor as animal model,the tumor targeting in vivo was determined by spectrophotofluorometer through the loaded calcein nanoparticles.The tumor inhibition information of F-BSANP-5-FU and BSANP-5-FU were evaluated by intraperitoneal injection, Original data was input to computer and was analyzed by the statistical software SPSS11.5.Inhibition rate (%)=( average tumor weight in contrast group - average tumor weight in dosage group) / average tumor weight in contrast group×100%; spleen and thymus index calculated as follows: spleen or thymus index = spleen or thymus weight (mg ) / mouse weight (g),the tumor of each group was made pathological section and the information of tumor suppression was observed .
Results: The coupling ratio of about 4 of folate-bovine serum albumin was synthesized with the most optimized synthesis conditions,that is a bovine serum albumin was coupled with 4 folates; 5-FU melting point was 283℃～284℃,5-FUA melting point was 276℃～278℃, yield was 81.37%;5-FUA active ester melting point was 255℃～257℃, yield was 83.98%;The results of ultraviolet、infrared、differential thermal analysis、MALDI-TOF MS showed that compounds were synthesized successfully;The nanoparticles whose particle size was less than 200nm were prepared successfully, and the size was uniform, the shape was integrated;The tumor targeting in vivo experiments showed that the equal doses of different groups of calcein were given by intraperitoneal injection,the accumulation of the content of calcein were different in vivo (P <0.05);The content of loaded calcein the BSANP-5-FU group (A group) reached its peak at 2.0h, and then declined in tumor tissues, The content of loaded calcein of Folate-BSANP-5-FU group (B group) reached its peak at 0.5h, and then declined sharply in tumor tissues . The content of calcein of liver and kidney tissues were less than the tumor tissue, but higher than the heart、spleen、lung tissue.The peak value of B group was significantly higher than A Group; The level of animal pharmacodynamic experiments showed that the tumor growth was the slowest in the tumor-bearing mice by intraperitoneal injection of F-BSANP-5-FU high-dose group, the highest inhibition rate reached 47.19%, the inhibition rate of F-BSANP-5-FU was significantly higher than BSANP-5-FU, after adding 1 mmol ? L-1 exogenous folate, the inhibition rate of F-BSANP-5-FU was declined significantly. The inhibition rate 5-FU positive control group was higher than BSANP-5-FU.
Conclusions: F-BSA-5-FU was synthesized under mild conditions, and its nanoparticles had tumor targeting, the targeting was likely to pass the folate-mediated, F-BSA-5-FU had good the inhibition of tumor,and its was a potential anti-tumor drugs that can be targeted at tumor with the high expression of folate receptor.
引文
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[42]陈雪岚,许杨,吴成钢.赭曲霉毒素A的酶联免疫检测-Ⅰ.抗原的制备[J].卫生研究,2002,31(1):53-55.
[43]韩克起,顾伟,胡侠,等.小鼠肝癌原位移植模型的建立及生物学特性[J].技术与方法,2004,2(5):372-374.
[44]李晓峰,叶韵斌,陈强,等.BABL/C×C57BL/6杂交F1代小鼠H22肝癌实体瘤模型的建立[J].福建医科大学学报,2006,40(4):387-390.
[45]梁朝霞.小鼠肿瘤模型的研究进展[J].国外医学卫生学分册,2004,31(4):212-216.
[46]刘军须,刘福英,蔡月花,等.不同来源H22细胞株的生物学特性比较研究[J].中国实验动物学报,2002,10(1):43-46.
[47]张阳德,吴泽建,龚连生,等.半乳糖化白蛋白磁性阿霉素纳米粒在正常大鼠体内的分布特征[J].中国医学工程,2004,12(4):5-7.
[48]万丹晶,钟高仁,徐培康,等.99Tcm-DTPA-Folate的制备及其荷瘤裸鼠体内分布[J].核技术,2004,27(4):293-296.
[49]张阳德,李玉坤,李浩,等.荧光分光光度法检测半乳糖化白蛋白磁性阿霉素纳米粒在大鼠体内分布的研究[J].中国医学工程,2004,12(4):26-30.
[50]苏华,胡晋红,李凤前.白蛋白纳米粒的制备工艺及靶向性研究进展[J].中国药学杂志,2005,40(9):641-644.
[51]毕芳,陶文沂,陆震鸣.白首乌提取物对小鼠肝癌细胞H22的抑制作用[J].中成药,2007,29(11):1586-1590.
[52] Lu Y J,Low P S.Folate-mediated delivery of macromolecular anticancer therapeutic agents[J].Advanced Drug Delivery Reviews,2002,54(5):675-693.
[53] Masaaki H,Masahiro M.Process for producing a high purity2, 4-dihydmxydipl1enyl sulfone [P]. EP: 755920,1997-01-29.
[54]周洁,曾甫清,高翔,等.三氧化二砷白蛋白纳米微球的制备及体外释药特性[J].中国新药杂志,2005,14(1):54-57.
[1]罗毅,卓仁禧.高分子抗肿瘤药物的研究[J].药学进展,1995,19(3):135-139.
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[3]杨亚楠,尹静波,刘芳.高分子材料在药物控制释放方面的应用[J].吉林工学院学报,2001,22(3):38-40.
[4] Chistie RJ,Grainger DW.Design strategies to improve macromoleuculardelivery constructs[J].Advanced Drug Delivery Reviews,2003,55:421-437.
[5] Giammona G,Caavallaro G.Studies of macromolecular prodrugs of zidovudine[J].Advance Drug Delivery Reviews,1999,39:153-164.
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[16]李和平,阮建明,蔡红革,等.壳聚糖-5-氟尿嘧啶高分子前药的合成与表征[J].长沙理工大学学报(自然科学版),2005,2(4):84-88.
[17]张静夏,潘仕荣,王琴梅,等.5-氟尿嘧啶-[N-(2-羟乙基)-L-谷酰胺]的合成及缓释性能研究[J].广州化学,2002,27(2):1-5.
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[8] Kondo SI,Shouichi H,Masayuki K, et al. Syntheses of novel hybrid polymeric prodrugs prepared by mechanochemical polymerization and the nature of their drug release [J].Drug Delivery Syst,1997,12(6):397-402.
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[13]周念恩,陈衢生,卓仁禧.高分子药物研究Ⅷ:二羟烷基嘧啶与N ,N′-双(2-氯乙基)二氯磷酰胺缩聚物的合成及其抗肿瘤活性研究[J].高分子学报,1987,(6):417-422.
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[17]张静夏,潘仕荣,王琴梅,等.5-氟尿嘧啶-[N-(2-羟乙基)-L-谷酰胺]的合成及缓释性能研究[J].广州化学,2002,27(2):1-5.
[18]朱卡琳,汤谷平,陈启琪,等.5-氟尿嘧啶-聚α,β-(2-羟乙基)-DL-天冬酰胺的合成及体内释放的研究[J].药学学报,33(12):906-909.
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[21] Marieta N,Etienne HS,Leonard WS,et al. Polymeric prodrugs of 5-fluorouracil[J].J Control Rel,1997,48:165-178.
[22] Marieta N,Etienne HS,Leonard WS.Macromolecular prodrugs of 5-fluorouracil 2: Enzymatic degradation [J].J Control Rel,1996,39:79-92.
[23]胡喜钢,汪森明,张起兴,等.以磺胺嘧啶为载体的氟尿嘧啶导向药物的合成[J].第一军医大学学报,2002,22(11):1042-1044.
[24]常军,郑玉斌,段久芳,等.含5-氟尿嘧啶的聚乙二醇酯的制备与表征[J].现代化工,2006,26(3):40-42.
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[31] Lee RJ,Wang S,Loe PS.Measurement of endosome pH following folate receptor-mediated endocytesis[J].Biochim Biophys Acta,l996,l3(12): 237-242.
[32]张良珂,侯世祥,毛声俊,等.叶酸偶联自蛋白纳米粒肿瘤细胞靶向性研究[J].四川大学学报(医学版),2004,35(2):165-168.
[33] Lee RJ,Low PS.Delivery of Liposomes into Cultured KB Cell via Folate Receptor-mediated Endocytosis[J].J Biolo Chem,1994,269(5):3198-3204.
[34] Cho KC,Jiong JH.Folate receptor-mediated intracellular delivery of recombinant caspase-3 for inducing apoptosis[J]. Journal of Controlled Release,2005,108:121-131.
[35] Weber C,Coester C.Desolvation process and surface characterisation of protein nanoparticles[J].Int J Pharm,2000,194:91-102.
[36] Leamon CP,Low PS.Folate-mediated targeting:from diagnostics to drug and gene delivery[J].Research focus,2001,6(1):44-58
[37] Jones GR,Tavish KM,Rice J.Vitamin-mediated targeting as a potential mechanism to increase drug uptake by tumours[J].Journal of Inorganic Biochemistry,2004,98:1652-1633.
[38] Leamon CP,Reddy JA.Folate-targeted chemotherapy[J].Adv Drug DelivRev,2004,56:1127-1141.
[39]吕会田,乐加昌,陈存社.克伦特罗小分子半抗原偶联率的测定方法研究[J].饲料工业,2005,36(3):57-59.
[40]蔡春,苏敏,杨健,等.肝靶向前药半乳糖化人血清白蛋白氟尿嘧啶偶联物的合成及靶向作用[J].中国药学杂志,2006,4l(10):786-789.
[41]扈靖,刘彦钦,韩士田.5-氟尿嘧啶-1-基乙酸合成方法的改进[J].化学试剂,2005,27(8):500-509.
[42]陈雪岚,许杨,吴成钢.赭曲霉毒素A的酶联免疫检测-Ⅰ.抗原的制备[J].卫生研究,2002,31(1):53-55.
[43]韩克起,顾伟,胡侠,等.小鼠肝癌原位移植模型的建立及生物学特性[J].技术与方法,2004,2(5):372-374.
[44]李晓峰,叶韵斌,陈强,等.BABL/C×C57BL/6杂交F1代小鼠H22肝癌实体瘤模型的建立[J].福建医科大学学报,2006,40(4):387-390.
[45]梁朝霞.小鼠肿瘤模型的研究进展[J].国外医学卫生学分册,2004,31(4):212-216.
[46]刘军须,刘福英,蔡月花,等.不同来源H22细胞株的生物学特性比较研究[J].中国实验动物学报,2002,10(1):43-46.
[47]张阳德,吴泽建,龚连生,等.半乳糖化白蛋白磁性阿霉素纳米粒在正常大鼠体内的分布特征[J].中国医学工程,2004,12(4):5-7.
[48]万丹晶,钟高仁,徐培康,等.99Tcm-DTPA-Folate的制备及其荷瘤裸鼠体内分布[J].核技术,2004,27(4):293-296.
[49]张阳德,李玉坤,李浩,等.荧光分光光度法检测半乳糖化白蛋白磁性阿霉素纳米粒在大鼠体内分布的研究[J].中国医学工程,2004,12(4):26-30.
[50]苏华,胡晋红,李凤前.白蛋白纳米粒的制备工艺及靶向性研究进展[J].中国药学杂志,2005,40(9):641-644.
[51]毕芳,陶文沂,陆震鸣.白首乌提取物对小鼠肝癌细胞H22的抑制作用[J].中成药,2007,29(11):1586-1590.
[52] Lu Y J,Low P S.Folate-mediated delivery of macromolecular anticancer therapeutic agents[J].Advanced Drug Delivery Reviews,2002,54(5):675-693.
[53] Masaaki H,Masahiro M.Process for producing a high purity2, 4-dihydmxydipl1enyl sulfone [P]. EP: 755920,1997-01-29.
[54]周洁,曾甫清,高翔,等.三氧化二砷白蛋白纳米微球的制备及体外释药特性[J].中国新药杂志,2005,14(1):54-57.
[1]罗毅,卓仁禧.高分子抗肿瘤药物的研究[J].药学进展,1995,19(3):135-139.
[2] Ohya Y,Ouchi T.Macromolecular prodrug [J].Prog Polym Sci,1995,20:211-257.
[3]杨亚楠,尹静波,刘芳.高分子材料在药物控制释放方面的应用[J].吉林工学院学报,2001,22(3):38-40.
[4] Chistie RJ,Grainger DW.Design strategies to improve macromoleuculardelivery constructs[J].Advanced Drug Delivery Reviews,2003,55:421-437.
[5] Giammona G,Caavallaro G.Studies of macromolecular prodrugs of zidovudine[J].Advance Drug Delivery Reviews,1999,39:153-164.
[6] Maeda H.Smancs and polymer-conjugated macromolecular drugs:advantages in cancer chemotherapy[J].Advanced Drug Delivery Reviews,2001,46:169-185.
[7] Pitha J.Vinyl polymer analogue of nucleic acids [J].Polymer,1977, 18:425-430.
[8] Kondo SI, Shouichi H, Masayuki K, et al. Syntheses of novel hybrid polymeric prodrugs prepared by mechanochemical polymerization and the nature of their drug release [J].Drug Delivery Syst,1997,12(6):397-402.
[9] Kondo SI,Shouichi H,Masayuki K,et al.Mechanochemical solid-state polymerization.Ⅷ:novel composition polymeric prodrugs prepared by mechanochemical polymerization in the presence of pharmaceutical aids [J]. Chem Pharm Bull,1998,52(11):1302-1306.
[10] Kondo SI,Shouichi H.Mechanochemical solid-state polymerization.Ⅺ:effect of water-insoluble pharmaceutical aids on drugs from mechanically synthesized polymeric prodrugs [J].Chem Pharm Bull,2004,46(4):669-673.
[11]王尊元,马臻.含5-氟尿嘧啶的高分子抗肿瘤药物的研究进展[J].中国医药工业杂志,2000,31(4):186-190.
[12]罗毅,卓仁禧,范昌烈.含5-氟尿嘧啶生物可降解聚磷酰胺的合成极其抗肿瘤活性研究[J].高等学校化学学报,1994,15(5):767-770.
[13]周念恩,陈衢生,卓仁禧.高分子药物研究Ⅷ.二羟烷基嘧啶与N ,N′-双(2-氯乙基)二氯磷酰胺缩聚物的合成及其抗肿瘤活性研究[J].高分子学报,1987,(6):417-422.
[14]杨福顺,卓仁禧.侧链含5-氟尿嘧啶甲壳胺的合成极其抗肿瘤活性的研究[J].高分子学报,1990,3(3):332-338.
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