包载表柔比星的生物素化泊洛沙姆聚合物胶束的构建与体内外初步评价
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摘要
目的:把生物素接枝到泊洛沙姆上制备包载表柔比星(EPI)的生物素-泊洛沙姆(BP)胶束。方法:分别对包载EPI的BP胶束的粒径、Zeta电位、表面形态、载药量、包封率及药物释放等方面进行考察和表征。用骨髓白血病HL-60细胞株评估包载EPI的BP胶束的体外细胞毒性。裸鼠皮下注射HL-60细胞建立肿瘤模型,考察包载EPI的BP胶束抑制肿瘤的影响。结果:这些纳米粒子的尺寸约为100 nm。荧光显微镜观察得出生物素结合的胶束有助于细胞摄取。抑制肿瘤体积增长的顺序:包载EPI的BP胶束>包载EPI的单胺封端的泊洛沙姆(MATP)胶束>包载EPI的泊洛沙姆胶束>单纯EPI。结论:与非靶向胶束相比,BP胶束表现出显著的的抗肿瘤活性和低毒性。拥有增强通透性和滞留性(EPR)效应和肿瘤靶向两大优势,BP胶束可能发展成为一个新的运载化疗药物的途径。尚需进一步进行其他细胞实验和大动物实验证明该肿瘤靶向技术。
Objective: To prepare biotin-poloxamer( BP) conjugate micelles for epirubicin through biotin conjugated on poloxamer. Methods: Epirubicin( EPI) was encapsulated in BP micelles. The EPI-loaded BP micelles were characterized by its particle size,zeta potential,surface morphology,as well as the efficiency of drug loading and drug encapsulation and drug release. Marrow leukemia HL-60 cells were used to evaluate the cell cytotoxicity of EPI-loaded BP micelles in vitro. The tumor model in nude mice was established through the subcutaneous injection of HL-60 cells,and then the inhibitory effect of EPI-loaded BP micelles on tumor volume growth was investigated. Results: It was found that the average particle size of EPI-loaded BP micelles was about 100 nm. In addition,the enhanced cellular uptake ability of EPI-loaded BP micelles was proved by fluorescence microscope observation. The efficiency order of the tumor volume growth inhibition was: EPI-loaded BP micelles > EPI-loaded MATP micelles > EPI-loaded poloxamer micelles >EPI. BP micelles showed significant antitumor activity and low toxicity when compared with the non-targeted micelles. Conclusion:With the advantages of EPR effect and tumor-targeting potential,BP conjugate micelles might be developed as a new system for chemotherapeutics. However,the tumor targeting technique should be demonstrated further by the other cell experiments and large animal experiments.
Objective: To prepare biotin-poloxamer( BP) conjugate micelles for epirubicin through biotin conjugated on poloxamer. Methods: Epirubicin( EPI) was encapsulated in BP micelles. The EPI-loaded BP micelles were characterized by its particle size,zeta potential,surface morphology,as well as the efficiency of drug loading and drug encapsulation and drug release. Marrow leukemia HL-60 cells were used to evaluate the cell cytotoxicity of EPI-loaded BP micelles in vitro. The tumor model in nude mice was established through the subcutaneous injection of HL-60 cells,and then the inhibitory effect of EPI-loaded BP micelles on tumor volume growth was investigated. Results: It was found that the average particle size of EPI-loaded BP micelles was about 100 nm. In addition,the enhanced cellular uptake ability of EPI-loaded BP micelles was proved by fluorescence microscope observation. The efficiency order of the tumor volume growth inhibition was: EPI-loaded BP micelles > EPI-loaded MATP micelles > EPI-loaded poloxamer micelles >EPI. BP micelles showed significant antitumor activity and low toxicity when compared with the non-targeted micelles. Conclusion:With the advantages of EPR effect and tumor-targeting potential,BP conjugate micelles might be developed as a new system for chemotherapeutics. However,the tumor targeting technique should be demonstrated further by the other cell experiments and large animal experiments.
引文
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27 Azade T,Rassoul D,Faranak SN,et al.Use of biotin targeted methotrexate-human serum albumin conjugated nanoparticles to enhance methotrexate antitumor efficacy[J].Int J Nanomedicine,2011,6:1863-1874
28 Yoo MK,Cho KY,Song HH,et al.Release of ciprofloxacin from chondroitin 6-sulfate-graft-poloxamer hydrogel in vitro for ophthalmic drug delivery[J].Drug Dev Ind Pharm,2005,31(4-5):455-463
29 Zhao YZ,Sun CZ,Lu CT,et al.Characterization and anti-tumor activity of chemical conjugationof doxorubicin in polymericmicelles(DOX-P)in vitro[J].Cancer Lett,2011,311(2):187-194
2 Batrakova EV,Li S,Brynskikh AM,et al.Effects ofpluronic and doxorubicin on drug uptake,cellular metabolism,apoptosisand tumor inhibition in animal models of MDR cancers[J].J Control Release,2010,143(3):290-301
3 Danquah MK,Zhang XA,Mahato RI.Extravasation of polymeric nanomedicinesacross tumor vasculature[J].Adv Drug Deliv Rev,2011,63(8):623-639
4 Kabanov A,Nazarova I,Astafieva I,et al.Micelle formation and solubilization of fluorescentprobes in poly(oxyethylene-boxypropilene-boxyethylene)[J].Macromolecules,1995,28(7):2303-2314
5 Croy SR,Kwon GS.Polymeric micelles for drug delivery[J].Curr Pharm Des,2006,12(36):4669-4684
6 Oerlemans C,Bult W,Bos M,et al.Polymericmicelles in anticancer therapy:targeting,imaging and triggered release[J].Pharm Res,2010,27(12):2569-2589
7 Schmolka IR.Poloxamers in the pharmaceutical industry.Polymers for controlled drug delivery[M].Boca Raton,FL:CRC Press,1991:189-214
8 Patil YB,Toti US,Khdair A,et al.Single-step surface functionalizationof polymeric nanoparticles for targeted drug delivery[J].Biomaterials,2009,30(5):859-866
9 Lu CT,Zhao YZ,Gao HS,et al.Comparing encapsulation efficiency andultrasound-triggered release for protein between phospholipidbasedmicrobubbles and liposomes[J].J Microencapsul,2010,27(2):115-121
10 Dinarvand R,Sepehri N,Manoochehri S,et al.Polylactideco-glycolide nanoparticles for controlled delivery of anticancer agents[J].Int J Nanomed,2011,6:877-895
11 Zhao YZ,Li X,Lu CT,et al.Experiment on the feasibility of using modified gelatin nanoparticlesas insulin pulmonary administration system for diabetes therapy[J].Acta Diabetol,2012,49(4):315-325
12 Chen S,Zhao X,Chen J,et al.Mechanism-based tumor-targeting drug delivery system validation ofefficient vitamin receptor-mediated endocytosis and drug release[J].Bioconjug Chem,2010,21(5):979-987
13 Swanson SD,Kukowska-Latallo JF,Patri AK,et al.Targeted gadoliniumloaded dendrimernanoparticles for tumor-specific magnetic resonance contrast enhancement[J].Int J Nanomed,2008,3(2):201-210
14 Zhao D,Zhao X,Zu Y,et al.Preparation,characterization,and in vitro targeted delivery of folate-decorated paclitaxelloadedbovine serum albumin nanoparticles[J].Int J Nanomed,2010,16(5):669-677
15 Li X,Tian X,Zhang J,et al.In vitro andin vivo evaluation of folate receptor-targeting amphiphilic copolymermodifiedliposomes loaded with docetaxel[J].Int J Nanomed,2011,6:1167-1184
16 Saltan N,Kutlu HM,Hür D,et al.Interaction of cancer cells withmagnetic nanoparticles modified by methacrylamido-folic acid[J].Int J Nanomed,2011,6:477-484
17 Xie J,Shen Z,Li KCP,et al.Tumor angiogenic endothelial cell targetingby a novel integrin-targeted nanoparticle[J].Int J Nanomed,2007,2(3):479-485
18 Na K.Self-assembled nanoparticles of hydrophobically-modified polysaccharidebearing vitamin H as a targeted anti-cancer drug deliverysystem[J].Eur J Pharm Sci,2003,18(2):165-173
19 Patil Y,Sadhukha T,Ma L,et al.Nanoparticle-mediated simultaneousand targeted delivery of paclitaxel and tariquidar overcomes tumor drugresistance[J].J Control Release,2009,136(1):21-29
20 Yang W,Cheng Y,Xu T,et al.Targeting cancer cellswith biotin dendrimer conjugates[J].Eur J Med Chem,2009,44(2):862–868
21 Taheri A,Dinarvand R,Nouri FS,et al.Use of biotin targeted methotrexatehumanserum albumin conjugated nanoparticles to enhancemethotrexate antitumor efficacy[J].Int J Nanomed,2011,6:1863-1874
22 Duncan R.The dawning era of polymer therapeutics[J].Nat Rev Drug Discov,2003,2(5):347-360
23 Broz P,Benito SM,Saw C,et al.Cell targeting by a generic receptortargetedpolymer nanocontainer platform[J].J Control Release,2005,102(2):475-488
24 Letchford K,Burt H.A review of the formation and classification of amphiphilicblock copolymer nanoparticulate structures:Micelles,nanospheres,nanocapsules and polymersomes[J].Eur J Pharm Biopharm,2007,65(3):259-269
25 Pulkkinen M,Pikkarainen J,Wirth T,et al.Three-step tumor targeting ofpaclitaxel using biotinylated PLA-PEG nanoparticles and avidin-biotintechnology:formulation development and in vitro anticancer activity[J].Eur J Pharm Biopharm,2008,70(1):66-74
26 Zhao YZ,Lu CT,Li XK,et al.Improving the cardio protective effect of a FGF inischemic myocardium with ultrasound-mediated cavitation of heparinmodified microbubbles:preliminary experiment[J].J Drug Target,2012,20(7):623-631
27 Azade T,Rassoul D,Faranak SN,et al.Use of biotin targeted methotrexate-human serum albumin conjugated nanoparticles to enhance methotrexate antitumor efficacy[J].Int J Nanomedicine,2011,6:1863-1874
28 Yoo MK,Cho KY,Song HH,et al.Release of ciprofloxacin from chondroitin 6-sulfate-graft-poloxamer hydrogel in vitro for ophthalmic drug delivery[J].Drug Dev Ind Pharm,2005,31(4-5):455-463
29 Zhao YZ,Sun CZ,Lu CT,et al.Characterization and anti-tumor activity of chemical conjugationof doxorubicin in polymericmicelles(DOX-P)in vitro[J].Cancer Lett,2011,311(2):187-194