目的 利用聚(2-乙基-2-噁唑啉)-胆固醇碳酸甲酯[poly(2-ethyl-2-oxazoline)-cholesteryl methyl carbonate,PEOz-CHMC,简称PC]构建多西紫杉醇(docetaxel,DOC)纳米胶束,并对其进行性质考察。方法 利用芘荧光探针法测定PC的临界胶束浓度。利用薄膜分散法制备DOC-PC胶束(DOC-PC micelles,DOC-M),对DOC-M的粒径、形态、包封率等进行表征。采用透析法考察DOC-M的药物释放,并模拟肿瘤微环境考察DOC-M的稳定性和pH敏感性。通过MTT法评价DOC-M对体外HeLa细胞的抑制作用。利用流式细胞仪定量观察胶束对HeLa细胞的摄取情况。结果 PC的临界胶束质量浓度为9.26 μg·mL-1(4.63×10-6mol·L-1)。DOC-M的粒径小于130 nm,外观呈类球形,分布均匀,Zeta电位为(-7.32±0.98)mV。X-射线粉末衍射(XRD)和红外光谱(IR)结果表明,DOC被成功包封于胶束中,DOC-M的包封率为(80.55±2.44)%。体外药物释放和胎牛血清稳定性结果实验表明,DOC-M的pH敏感性和稳定性良好。细胞抑制实验结果表明,在微酸条件下DOC-M的细胞抑制作用更强。细胞摄取实验分析,DOC-M具有较低的毒性,显著地促进药物的细胞摄取。结论 DOC-M表现出良好的稳定性和pH敏感性,以及较低毒性和较好的载药能力,有望成为药物递送的良好载体。
Abstract
OBJECTIVE To construct docetaxel(DOC) nano-micelles using poly(2-ethyl-2-oxazoline)-cholesteryl methyl carbonate(PEOz-CHMC, PC).METHODS The critical micelle concentration of PC was measured by the hydrazine fluorescence probe method, and DOC-PC micelles(DOC-micelles, DOC-M) were prepared by the thin-film dispersion method. The particle size,morphology,and encapsulation efficiency of DOC-M were characterized.Drug release of DOC-M was evaluated by the dialysis method.Stability and pH-sensitivity of micelles were investigated by simulating the tumor microenvironment.In vitro inhibitory effect of DOC-M was measured by the MTT method in Hela cells. Cell uptake experiments were performed to investigate the uptake of DOC-M in Hela cells by flow cytometry.RESULTS CMC of PC was 9.26 μg·mL-1(4.63×10-6mol·L-1).The appearance of DOC-M showed a regular spherical shape, and the particle size of DOC-M was less than 130 nm and the size distribution was uniform. The Zeta potential and the encapsulation efficiency of DOC-M were (-7.32±0.98) mV and(80.55±2.44)%, respectively.The results of XRD and IR showed that DOC was successfully encapsulated into micelles. In vitro drug release and fetal bovine serum stability results indicated that both the stability and pH-sensitivity of DOC-M were excellent. The results of cytostatic assay suggested that the cell inhibitory effect of DOC-M was stronger under slightly acidic conditions. Cellular uptake analysis showed that DOC-M had low toxicity and significantly promoted the cellular uptake of drugs.CONCLUSION DOC-M exhibits good stability and pH sensitivity, as well as low toxicity and good drug loading capacity, and is expected to become a good carrier for drug delivery.
关键词
胶束 /
pH敏感 /
聚(2-乙基-2-噁唑啉)-胆固醇碳酸甲酯 /
多西紫杉醇
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Key words
micelle /
pH-sensitivity /
poly(2-ethyl-2-oxazoline)-cholesterol methyl carbonate /
docetaxel
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中图分类号：
R944
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参考文献
[1] ENGELS F K, MATHOT R A A, VERWEI J. Alternative drug formulations of docetaxel: a review[J]. Anti-Cancer Drugs, 2007, 18(2): 95-103.
[2] HOANG B, ERNSTING M J, MURAKAMI M, et al. Docetaxel-carboxymethylcellulose nanoparticles display enhanced anti-tumor activity in murine models of castration-resistant prostate cancer[J]. Int J Pharm, 2014, 471(1-2): 224-233.
[3] WANG C H, WANG C H, HSIUE G H. Polymeric micelles with a pH-responsive structure as intracellular drug carriers[J]. J Controlled Release, 2005, 108(1): 140-149.
[4] WANG C H, HSIUE G H. Synthesis and characterization of temperature-and pH-sensitive hydrogels based on poly(2-ethyl-2-oxazoline) and poly(D, L-lactide)[J]. J Polym Sci Part A Polym Chem, 2002, 40(8): 1112-1121.
[5] WANG C H, HSIUE G H. New amphiphilic poly(2-ethyl-2-oxazoline)/poly(L-lactide) triblock copolymers[J]. Biomacromolecules, 2003, 4(6): 1487-1490.
[6] XU H, HU M, YU X, et al. Design and evaluation of pH-sensitive liposomes constructed by poly(2-ethyl-2-oxazoline)-cholesterol hemisuccinate for doxorubicin delivery[J]. Eur J Pharm Biopharm, 2015, 91: 66-74.
[7] XU H, ZHANG W, LI Y, et al. The bifunctional liposomes constructed by poly(2-ethyl-oxazoline)-cholesteryl methyl carbonate: an effectual approach to enhance liposomal circulation time, pH-sensitivity and endosomal escape [J]. Pharm Res, 2014, 31(11): 3038-3050.
[8] OERLEMANS C, BULT W, BOS M, et al. Polymeric micelles in anticancer therapy: targeting, imaging and triggered release[J]. Pharm Res, 2010, 27(12): 2569-2589.
[9] PEER D, KARP J M, HONG S, et al. Nanocarriers as an emerging platform for cancer therapy[J]. Nat Nanotechnol, 2007, 2(12): 751-760.
[10] SINGH S. Nanomedicine-nanoscale drugs and delivery systems[J]. J Nanosci Nanotechnol, 2010, 10(12): 7906-7918.
[11] KATAOKA K, HARADA A, NAGASAKI Y. Block copolymer micelles for drug delivery: design, characterization and biological significance[J]. Adv Drug Deliv Rev, 2012, 64: 37-48.
[12] BARREIRO-IGLESIAS R, BROMBERG L, TEMCHENKO M, et al. Solubilization and stabilization of camptothecin in micellar solutions of pluronic-g-poly(acrylic acid) copolymers[J]. J Controlled Release, 2004, 97(3): 537-549.
[13] DANHIER F, FERON O, PRéAT V. To exploit the tumor microenvironment: passive and active tumor targeting of nanocarriers for anti-cancer drug delivery[J]. J Controlled Release, 2010, 148(2): 135-146.
[14] SHEU M T, JHAN H J, SU C Y, et al. Codelivery of doxorubicin-containing thermosensitive hydrogels incorporated with docetaxel-loaded mixed micelles enhances local cancer therapy[J]. Colloid Surface B, 2016, 143: 260-270.
[15] ZHANG C Y, YANG Y Q, HUANG T X, et al. Self-assembled pH-responsive MPEG-b-(PLA-co-PAE) block copolymer micelles for anticancer drug delivery[J]. Biomaterials, 2012, 33(26): 6273-6283.
[16] WANG D, ZHOU Y, LI X, et al. Mechanisms of pH-sensitivity and cellular internalization of PEOz-b-PLA micelles with varied hydrophilic/hydrophobic ratios and intracellular trafficking routes and fate of the copolymer[J]. ACS Appl Mater Interfaces, 2017, 9(8): 6916-6930.
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脚注
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基金
江西中医药大学现代中药制剂教育部重点实验室开放基金项目资助(TCM-201913);辽宁省教育厅高等学校自然科学类基本科研(面上)项目资助(LJKZ0981)
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