卡铂@葡聚糖纳米载体的自组装/聚合一步法制备及生物应用
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摘要
在接枝共聚辅助自组装(GISA)制备葡聚糖纳米载体的过程中,利用丙烯酸单体与卡铂之间的非共价键作用,使得卡铂参与到葡聚糖纳米载体的形成中,从而一步实现了卡铂@葡聚糖纳米载体的制备,并使用肿瘤还原性环境敏感的二硫键来交联纳米载体,得到了对肿瘤还原环境响应的纳米药物载体.对纳米药物载体的结构、粒径及形貌进行表征,结果显示,纳米药物载体粒径为(92±0. 2) nm,Zeta电位为(-8±0. 3) e V.通过体外药物释放研究发现,在还原性环境中,载体可持续72 h释放药物,最大释放量达80%.细胞摄取实验表明负载卡铂的纳米药物载体可在4 h内高效地进入细胞核;其半抑制浓度(IC_(50))为25. 32μg/mL,达到和相同浓度游离卡铂相仿的促肿瘤细胞凋亡效果.此一步法所制备的卡铂@葡聚糖纳米载体具有良好的生物应用前景.
We introduced disulfide bond in the crosslinking points of the carboplatin@dextran to endow the reducing environmental sensitivity to the nanocarriers. Its structure,particle size and morphology of the nanocarrier were characterized by means of dynamic light scattering( DLS),1 H NMR and FTIR. The results showed that the size of the nanocarrier was( 92 ± 0. 2) nm. In vitro drug release studies disclosed that the carboplatin inside the nanocarrier can be released for 80% in 72 h in a reducing environment. The cell uptake experiments showed that the nanocarriers loaded with carboplatin could efficiently enter the cell nuclei within4 h. The half maximal inhibitory concentration( IC_(50)) of the carboplatin @ dextran nanocarrier was 25. 32μg/m L,which had similar cytotoxicity to the free carboplatin at the same concentration. The above results demonstrate that carboplatin @ dextran nanocarrier prepared by the one-step method has a good biomedical application prospect.
We introduced disulfide bond in the crosslinking points of the carboplatin@dextran to endow the reducing environmental sensitivity to the nanocarriers. Its structure,particle size and morphology of the nanocarrier were characterized by means of dynamic light scattering( DLS),1 H NMR and FTIR. The results showed that the size of the nanocarrier was( 92 ± 0. 2) nm. In vitro drug release studies disclosed that the carboplatin inside the nanocarrier can be released for 80% in 72 h in a reducing environment. The cell uptake experiments showed that the nanocarriers loaded with carboplatin could efficiently enter the cell nuclei within4 h. The half maximal inhibitory concentration( IC_(50)) of the carboplatin @ dextran nanocarrier was 25. 32μg/m L,which had similar cytotoxicity to the free carboplatin at the same concentration. The above results demonstrate that carboplatin @ dextran nanocarrier prepared by the one-step method has a good biomedical application prospect.
引文
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[2]Cleare M.J.,Developments in Pharmacology,Springer Netherlands,Berlin,1983,59-91
[3]Fuertes M.A.,Alonso C.,Pérez.,JoséM.,Chemical Reviews,2003,103(3),645-662
[4]Knox R.J.,Friedlos F.,Lydall D.A.,Roberts J.J.,Cancer Research,1986,46(4),1972-1979
[5]Ho G.Y.,Woodward N.,Coward J.I.,Critical Reviews in Oncology/hematology,2016,102,37-46
[6]Rosenberg B.,Camp L.V.,Krigas T.,Nature,1965,205(4972),698-699
[7]Zorbas H.,Keppler B.K.,Chembiochem.,2005,6(7),1157-1166
[8]Sousa G.F.D.,Wlodarczyk S.R.,Monteiro G.,Brazilian Journal of Pharmaceutical Science,2014,50(4),693-701
[9]Yonezawa A.,Masuda S.,Yokoo S.,Katsura T.,Inui K.,Journal of Pharmacology&Experimental Therapeutics,2006,319(2),879-886
[10]Shahzad M.M.K.,Berestein G.L.,Sood A.K.,Drug Resist.Update.,2009,12(6),148-152
[11]Fang J.,Nakamura H.,Maeda H.,Advanced Drug Delivery Reviews,2011,63(3),136-151
[12]Hatakeyama H.,Akita H.,Harashima H.,Advanced Drug Delivery Reviews,2011,63(3),152-160
[13]Ganta S.,Devalapally H.,Shahiwala A.,Amiji M.,Journal of Controlled Release,2008,126(3),187-204
[14]Los G.,Verdegaal E.,Noteborn H.P.,Ruevekamp M.,de Graeff A.,Meesters E.W.,ten BokkelHuinink D.,Mc Vie J.G.,Biochemical Pharmacology,1991,42(2),357-363
[15]Wang F.,Wang Y.C.,Dou S.,Xiong M.H.,Sun T.M.,Wang J.,ACS Nano,2011,5(5),3679-3692
[16]Sprowl J.A.,Ness R.A.,Sparreboom A.,Drug Metabolism&Pharmacokinetics,2013,28(1),19-27
[17]Hampel S.,Kunze D.,Haase D.,Krmer K.,Rauschenbach M.,Ritschel M.,Leonhardt A.,Thomas J.,Oswald S.,Hoffmann V.,Büchner B.,Nanomedicine,2008,3(2),175-182
[18]Balas M.,Constanda S.,Duma A.,Prodana M.,Hermenean A.,Pop S.,Demetrescu I.,Dinischiotu A.,Toxicology in Vitro,2016,37,189-200
[19]Arshad A.,Yang B.,Bienemann A.S.,Barua N.U.,Wyatt M.J.,Woolley M.,Johnson D.E.,Edler K.J.,Gill S.S.,PloS One,2015,10(7),1-16
[20]Alex A.T.,Joseph A.,Shavi G.,Rao J.V.,Udupa N.,Drug Delivery,2016,23(7),21-44
[21]Chaudhury A.,Das S.,Bunte R.M.,Chiu G.N.,International Journal of Nanomedicine,2012,7,739-751
[22]Ebrahimifar M.,Nili-Ahmadabadi A.,Akbarzadeh A.,Shahemabadi H.E.,Hasanzadegan M.,Moradi-Sardareh H.,Madadizadeh H.,Rezaee-Diyan J.,Indian Journal of Clinical Biochemistry,2017,32(2),230-234
[23]Zhang X.,Liu Y.,Kim Y.J.,Mac J.,Zhuang R.,Wang P.,RSC Advances,2017,7(32),19685-19693
[24]Malik N.,Wiwattanapatapee R.,Klopsch R.,Lorenz K.,Frey H.,Weener J.W.,Meijer E.W.,Paulus W.,Duncan R.,Journal of Controlled Release,2000,65(1),133-148
[25]Malik N.,Evagorou E.R.,Anticancer Drugs,1999,10(8),767-776
[26]Wang Y.,Wang L.,Chen G.,Gong S.,Macromolecular Bioscience,2017,17,1600292-1600301
[27]Zhu X.,Peng Y.,Qiu L.,Colloids and Surfaces B:Biointerfaces,2017,157,56-64
[28]Yu S.,Gan M.C.,Journal of Materials Chemistry,2004,14(18),2781-2786
[29]Potara M.,Nagy-Simon T.,Craciun A.M.,Suarasan S.,Licarete E.,Imre-Lucaci F.,Astilean S.,ACS Applied Materials&Interfaces,2017,9(38),32565-32576
[30]Tang M.H.,Dou H.J.,Sun K.,Polymer,2006,47(2),728-734
[31]Bachelder E.M.,Beaudette T.T.,Broaders K.E.,Dashe J.,Fréchet J.M.,Journal of the American Chemical Society,2008,130(32),10494-10495
[32]Liu W.Y.,Hao Y.Y.,Hu Y.,Chen W.C.,Dai J.,Journal of Cellulose Science and Technology,2015,23(1),60-65
[33]Du Z.,Zhang Y.,Li Z.J.,Chen H.,Wang Y.,Wang G.T.,Zou P.,Chen H.P.,Zhang Y.S.,Applied Surface Science,2017,392,312-320
[34]Cui L.L.,Liu J.M.,Fan H.L.,Xu X.W.,Yang M.,Xiao J.P.,Rare Metals,2007,26(3),276-279
[35]Zhang C.,Ping Q.N.,Zhang H.J.,Shen J.,Carbohydrate Polymers,2003,54(2),137-141
[36]Dou H.J.,Jiang M.,Peng H.S.,Chen D.Y.,Hong Y.,Angew.Chem.Int.Ed.,2003,42,1516-1519
[37]Kuppusamy P.,Li H.,Ilangovan G.,Cardounel A.J.,Zweier J.L.,Yamada K.,Krishna M.C.,Mitchell J.B.,Cancer Research,2002,62(1),307-312
[38]Meng F.H.,Hennink W.E.,Zhong Z.Y.,Biomaterials,2009,30(12),2180-2198