叶酸修饰聚酰胺-胺负载5-氟尿嘧啶纳米给药体系的制备及其体外性能研究
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摘要
使用化学键键合的方式分别将叶酸(FA)和5-氟尿嘧啶-1-基乙酸(FUA)偶联于树状大分子聚酰胺-胺(PAMAM)制备纳米载药体系FA-PAMAM-FUA,并结合单因素分析法筛选出最佳制备条件为FA∶PAMAM∶FUA的物质的量的比为10∶1∶450,p H值为6.用紫外可见光谱、红外光谱和纳米粒度仪对FA-PAMAM-FUA的光学特性、粒径大小和体外释药性能进行了表征.结果表明:优化条件下制备的FAPAMAM-FUA粒径呈正态分布,平均粒径为90.28±20.00 nm,分散度为0.258±0.002(n=3),体系载药量达28.45%,FA-PAMAM-FUA给药体系对药物具有一定的p H敏感特性以及缓释功能.采用MTT法考察了5-Fu、FA-PAMAM-FUA对乳腺癌细胞MCF-7的增殖抑制作用,结果表明两者对MCF-7细胞的增殖具有明显地抑制作用,剂量依赖性强,随浓度的增高,FA-PAMAM-FUA对癌细胞增殖抑制作用较5-Fu明显增强.
Drug delivery nanosystem FA-PAMAM-FUA was prepared through covalently coupling the amino group of polyamide amine( PAMAM) with the carboxylic group of folicacid( FA) and 5-fluorouracil acetic acid( FUA),respectively. The optimized mole ratio of FA ∶ PAMAM ∶ FUA was 10 ∶ 1 ∶ 450 and the optimized p H value was 6.The optical and release properties of as-prepared FA-PAMAM-FUA were characterized using UV-Visible spectra,infrared spectra,particle size and zeta potential. The average particle size and size dispersion of FA-PAMAM-FUA was 90. 28 ± 20. 00 nm and 0. 258 ± 0. 002( n = 3),respectively.The drug-loading content of FA-PAMAM-FUA was 28.45%.The in vitro release profile of FUA from FA-PAMAM-FUA demonstrated a low and sustained release behaviour.The MTT assay was used to assess the proliferation inhibition of 5-Fu and FA-PAMAMFUA on breast cancer cell MCF-7.The MTT results showed that both of them had a significant proliferation-inhibitory effect and a dose-dependent effect on the MCF-7 cells.And the results demonstrated that the resulting FA-PAMAM-FUA nanosystem were more potent in killing cancer cells as the spiked free 5-Fu or FA-PAMAM-FUA concentration were increased,compared to free 5-Fu.
Drug delivery nanosystem FA-PAMAM-FUA was prepared through covalently coupling the amino group of polyamide amine( PAMAM) with the carboxylic group of folicacid( FA) and 5-fluorouracil acetic acid( FUA),respectively. The optimized mole ratio of FA ∶ PAMAM ∶ FUA was 10 ∶ 1 ∶ 450 and the optimized p H value was 6.The optical and release properties of as-prepared FA-PAMAM-FUA were characterized using UV-Visible spectra,infrared spectra,particle size and zeta potential. The average particle size and size dispersion of FA-PAMAM-FUA was 90. 28 ± 20. 00 nm and 0. 258 ± 0. 002( n = 3),respectively.The drug-loading content of FA-PAMAM-FUA was 28.45%.The in vitro release profile of FUA from FA-PAMAM-FUA demonstrated a low and sustained release behaviour.The MTT assay was used to assess the proliferation inhibition of 5-Fu and FA-PAMAMFUA on breast cancer cell MCF-7.The MTT results showed that both of them had a significant proliferation-inhibitory effect and a dose-dependent effect on the MCF-7 cells.And the results demonstrated that the resulting FA-PAMAM-FUA nanosystem were more potent in killing cancer cells as the spiked free 5-Fu or FA-PAMAM-FUA concentration were increased,compared to free 5-Fu.
引文
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[26]唐国涛,曹轩,贺冬秀,等.聚酰胺-胺树枝状高分子/喜树碱复合物的制备及体外释放研究[J].南华大学学报(自然科学版),2011,25(2):79-83.
[27]KESHARWANI P,BANERJEE S,GUPTA U,et al.PAMAM dendrimers as promising nanocarriers for RNAi therapeutics[J].Materials today,2015,18(10):565-572.
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[2]徐鹏程.纳米靶向给药系统的抗肿瘤应用前景分析[J].内蒙古科技与经济,2016(8):86-89.
[3]章伟,赵林林,陈立伟,等.恶性肿瘤化疗后骨髓抑制的中医药防治进展[J].四川中医,2014,32(6):179-182.
[4]WILSON B,AMBIKA T V,PATEL R D,et al.Nanoparticles based on albumin:preparation,characterization and the use for 5-flurouracil delivery[J].International journal of biological macromolecules,2012,51(5):874-878.
[5]CHENG M,HE B,WAN T,et al.5-fluorouracil nanoparticles inhibit hepatocellular carcinoma via activation of the p53 pathway in the orthotopic transplant mouse model[J].Plo S One,2012,7(10):47115.
[6]FANG W.Chemotherapy in patient with colon cancer after renal transplantation:a case report with literature review[J].Medicine(Baltimore),2018,97(5):9678.
[7]GAO Z,LI Z,YAN J,et al.Irinotecan and 5-fluorouracil-coloaded,hyaluronic acid-modified layer-by-layer nanoparticles for targeted gastric carcinoma therapy[J].Drug design,development and therapy,2017,11:2595-2604.
[8]BWATANGLANG I B,MOHAMMAD F,YUSOF N A,et al.In vivo tumor targeting and anti-tumor effects of 5-fluororacil loaded,folic acid targeted quantum dot system[J].Journal of colloid and interface science,2016,480:146-158.
[9]WETTERGREN Y,CARLSSON G,ODIN E,et al.Pretherapeutic uracil and dihydrouracil levels of colorectal cancer patients are associated with sex and toxic side effects during adjuvant 5-Fluorouracil-based chemotherapy[J].Cancer,2012,118(11):2935-2943.
[10]孙维彤,陈倩倩,张娜,等.5-氟尿嘧啶前体药物研究进展[J].生命的化学,2014,34(2):269-274.
[11]赵晶晶,沈灵佳,周建平,等.纳米靶向给药系统载体材料的研究进展[J].药学进展,2015,39(3):161-169.
[12]李劲草,孙岚,姜爽,等.纳米药物释放系统在肿瘤组织中增强的透过与滞留效应及其影响因素[J].药学进展,2015,29(1):164-169.
[13]MAEDA H,BHARATE G Y,DARUWALLA J.Polymeric drugs for efficient tumor-targeted drug delivery based on EPR-effect[J].European journal of pharmaceutics and biopharma ceutics,2009,71(3):409-419.
[14]LEI X,LI K,LIU Y,et al.Co-delivery nanocarriers targeting folate receptor and encapsulating 2-deoxyglucose andα-tocopheryl succinate enhance anti-tumor effect in vivo[J].International journal of nanomedicine,2017,12:5701-5715.
[15]HU Y.Targeted nanosystems:advances in targeted dendrimers for cancer therapy[J].Nanomedicine,2016,12:309-316.
[16]李娜梅,杨飒,宁倩,等.半乳糖化壳聚糖-氟尿嘧啶偶合物纳米给药体系的制备、表征及体外性能研究[J].南华大学学报(自然科学版),2015,29(3):82-86;91.
[17]KANG T,LI F,BAIK S,et al.Surface design of magnetic nanoparticles for stimuli-responsive cancer imaging and therapy[J].Biomaterials,2017,136:98-114.
[18]SANTIAGO T,DEVAUX R S,KURZATKOWSKA K,et al.Surface-enhanced raman scattering investigation of targeted delivery and controlled release of gemcitabine[J].International journal of nanomedicine,2017,12:7763-7776.
[19]FRACZYK J,WALCZAK M,SZYMANSKI L,et al.Carbon nanotubes functionalized with folic acid attached via biomimetic peptide linker[J].Nanomedicine,2017,12(18):2161-2182.
[20]ALVAREZ-BERRIOS M P,VIVERO-ESCOTO J L.In vitro evaluation of folic acid-conjugated redox-responsive mesoporous silica nanoparticles for the delivery of cisplatin[J].International journal of nanomedicine,2016,11:6251-6265.
[21]MARCHETTI C,PALAIA I,GIORGINI M,et al.Targeted drug delivery via folate receptors in recurrent ovarian cancer:a review[J].Onco Targets and therapy,2014,7:1223-1236.
[22]MEKEDA C.Folate targeted agents for diagnostic and terapeutic uses[D].Chicago:Governors state university,2014.
[23]JI Z,LIN G,LU Q,et al.Targeted therapy of SMMC-7721 liver cancer in vitro and in vivo with carbon nanotubes based drug delivery system[J].Journal of colloid and interface science,2012,365:143-149.
[24]THAPA R K,CHOI J Y,POUDEL B K,et al.Receptor-targeted,drug-loaded,functionalized graphene oxides for chemotherapy and photothermal therapy[J].International journal of nanomedicine,2016,11:2799-2813.
[25]LI H,PIAO L,YU B,et al.Delivery of calf thymus DNA to tumor by folate receptor targeted cationic liposomes[J].Biomaterials,2011,32:6614-6620.
[26]唐国涛,曹轩,贺冬秀,等.聚酰胺-胺树枝状高分子/喜树碱复合物的制备及体外释放研究[J].南华大学学报(自然科学版),2011,25(2):79-83.
[27]KESHARWANI P,BANERJEE S,GUPTA U,et al.PAMAM dendrimers as promising nanocarriers for RNAi therapeutics[J].Materials today,2015,18(10):565-572.
[28]LI N M,YANG S,NING Q,et al.Fabrication of galactosylated chitosan-5-fluorouracil acetic acid based nanoparticles for controlled drug delivery[J].Journal of applied polymer science,2015,132(40):42625-42631.