双重pH敏感的聚合物-阿霉素偶联物胶束的制备与表征（英文）
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摘要
为了有效抑制肿瘤生长并降低抗肿瘤药物的毒副作用,本研究设计并构建了基于阿霉素(DOX)与聚(2-乙基-2-噁唑啉)-聚乳酸(PEOz-PLA)偶联物(PEOz-PLA-imi-DOX)的双重pH敏感聚合物胶束。成功合成了DOX与pH敏感的PEOz-PLA经酸敏感的安息香亚胺键偶联的PEOz-PLA-imi-DOX,并用1H NMR和薄层色谱进行了确证,芘荧光探针法测定其临界胶束浓度为(14.84±3.85)mg/L。由PEOz-PLA-imi-DOX经薄膜水化法制备的偶联物胶束(PP-DOX-PM)的粒径约为21 nm,载药量为1.67%。体外释放实验表明,PP-DOX-PM具有随着pH的降低而释放逐渐加快的pH依赖性释放行为,说明该胶束能识别正常生理环境和胞内的pH差异,预示该胶束具有胞内快速释药的特征。此外,PP-DOX-PM保持了DOX对MDA-MB-231细胞的毒性。因此,PP-DOX-PM胶束在抗肿瘤方面具有潜在的优势。
In the present study, we designed and fabricated pH-sensitive polymeric micelles based on the conjugate of poly(2-ethyl-2-oxazoline)-poly(D,L-lactide)(PEOz-PLA) with doxorubicin(PEOz-PLA-imi-DOX) to efficiently inhibit tumor cell growth. Hence, PEOz-PLA-imi-DOX was successfully synthesized by connecting DOX to the hydrophobic end of pH-sensitive PEOz-PLA via acid cleavable benzoic imine linker and characterized by 1 H NMR spectrum and thin layer chromatography. The critical micelle concentration of PEOz-PLA-imi-DOX was determined to be(14.84±3.85) mg/L. The conjugate micelles(denoted as PP-DOX-PM) formed by PEOz-PLA-imi-DOX using film-hydration method were characterized to have a nano-scaled size of about 21 nm in diameter, and the drug loading content was 1.67%. PP-DOX-PM showed pH-dependent drug release behavior with gradually accelerated release of DOX with decrease of pH value, illustrating the micelles' distinguishing feature of endo/lysosomal pH from physiological pH by accelerating drug release. As anticipated, PP-DOX-PM maintained the cytotoxicity of DOX against MDA-MB-231 cells. Collectively, PP-DOX-PM might have great potential for effective suppression of tumor growth.
In the present study, we designed and fabricated pH-sensitive polymeric micelles based on the conjugate of poly(2-ethyl-2-oxazoline)-poly(D,L-lactide)(PEOz-PLA) with doxorubicin(PEOz-PLA-imi-DOX) to efficiently inhibit tumor cell growth. Hence, PEOz-PLA-imi-DOX was successfully synthesized by connecting DOX to the hydrophobic end of pH-sensitive PEOz-PLA via acid cleavable benzoic imine linker and characterized by 1 H NMR spectrum and thin layer chromatography. The critical micelle concentration of PEOz-PLA-imi-DOX was determined to be(14.84±3.85) mg/L. The conjugate micelles(denoted as PP-DOX-PM) formed by PEOz-PLA-imi-DOX using film-hydration method were characterized to have a nano-scaled size of about 21 nm in diameter, and the drug loading content was 1.67%. PP-DOX-PM showed pH-dependent drug release behavior with gradually accelerated release of DOX with decrease of pH value, illustrating the micelles' distinguishing feature of endo/lysosomal pH from physiological pH by accelerating drug release. As anticipated, PP-DOX-PM maintained the cytotoxicity of DOX against MDA-MB-231 cells. Collectively, PP-DOX-PM might have great potential for effective suppression of tumor growth.
引文
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[2]Bae,Y.;Jang,W.D.;Nishiyama,N.;Fukushima,S.;Kataoka,K.Multifunctional Polymeric Micelles with Folate-Mediated Cancer Cell Targeting and p H-Triggered Drug Releasing Properties for Active Intracellular Drug Delivery.Mol.Biosyst.2005,1,242-250.
[3]Guo,X.;Shi,C.;Wang,J.;Di,S.;Zhou,S.p H-Triggered Intracellular Release from Actively Targeting Polymer Micelles.Biomaterials.2013,34,4544-4554.
[4]Yang,X.;Grailer,J.J.;Pilla,S.;Steeber,D.A.;Gong,S.Tumor-Targeting,p H-Responsive,and Stable Unimolecular Micelles as Drug Nanocarriers for Targeted Cancer Therapy.Bioconjug.Chem.2010,21,496-504.
[5]Wang,Y.;Luo,Q.;Sun,R.;Zha,G.;Li,X.;Shen,Z.;Zhu,W.Acid-Triggered Drug Release from Micelles Based on Amphiphilic Oligo(Ethylene Glycol)-Doxorubicin Alternative Copolymers.J.Mater.Chem.B.2014,2,7612-7619.
[6]Requejo,A.R.;Alastrue,A.A.;Cases,V.M.;Lopez,M.E.;England,R.;Vicent,M.J.;Moreno,M.V.Combined Polymer-Curcumin Conjugate and Ependymal Progenitor/Stem Cell Treatment Enhances Spinal Cord Injury Functional Recovery.Biomaterials.2017,113,18-30.
[7]Kyluik Price,D.L.;Li,L.;Scott,M.D.Comparative Efficacy of Blood Cell Immunocamouflage by Membrane Grafting of Methoxypoly(Ethylene Glycol)and Polyethyloxazoline.Biomaterials.2014,35,412-422.
[8]Wang,C.H.;Hsiue,G.H.Synthesis and Characterization of Temperature-and p H-Sensitive Hydrogels Based on Poly(2-Ethyl-2-Oxazoline)and Poly(D,L-Lactide).J.Pol.Sci.Poly.Chem.2002,40,1112-1121.
[9]Wang,Z.;Li,X.;Wang,D.;Zou,Y.;Qu,X.;He,C.;Deng,Y.;Jin,Y.;Zhou,Y.;Zhou,Y.;Liu,Y.Concurrently Suppressing Multidrug Resistance and Metastasis of Breast Cancer by Co-Delivery of Paclitaxel and Honokiol with p H-Sensitive Polymeric Micelles.Acta Biomater.2017,62,114-156.
[10]Li,J.;Zhou,Y.;Li,C.;Wang,D.;Gao,Y.;Zhang,C.;Zhao,L.;Li,Y.;Liu,Y.;Li,X.Poly(2-Ethyl-2-Oxazoline)-Doxorubicin Conjugate-Based Dual Endosomal p H-Sensitive Micelles with Enhanced Antitumor Efficacy.Bioconjug.Chem.2015,26,110-119.
[11]Qu,X.;Yang,Z.Benzoic-Imine-Based Physiologicalp H-Responsive Materials for Biomedical Applications.Chem.Asian.J.2016,11,2633-2641.
[12]Wang,D.;Zhou,Y.;Li,X.;Qu,X.;Deng,Y.;Wang,Z.;He,C.;Zou,Y.;Jin,Y.;Liu,Y.Mechanisms of PhSensitivity and Cellular Internalization of Peoz-B-Pla Micelles with Varied Hydrophilic/Hydrophobic Ratios and Intracellular Trafficking Routes and Fate of the Copolymer.ACS Appl.Mater.Interfaces.2017,9,6916-6930.
[13]Gao,Y.;Li,Y.;Li,Y.;Yuan,L.;Zhou,Y.;Li,J.;Zhao,L.;Zhang,C.;Li,X.;Liu,Y.Psma-Mediated Endosome Escape-Accelerating Polymeric Micelles for Targeted Therapy of Prostate Cancer and the Real Time Tracing of Their Intracellular Trafficking.Nanoscale.2014,7,597-612.
[14]Zhu,L.;Zhao,L.;Qu,X.;Yang,Z.p H-Sensitive Polymeric Vesicles from Coassembly of Amphiphilic Cholate Grafted Poly(L-Lysine)and Acid-Cleavable Polymer-Drug Conjugate.Langmuir.2012,28,11988-11996.
[15]Perrault,S.D.;Walkey,C.;Jennings,T.;Fischer,H.C.;Chan,W.C.W.Mediating Tumor Targeting Efficiency of Nanoparticles through Design.Nano Letters.2009,9,1909-1915.
[16]Kim,D.;Gao,Z.G.;Lee,E.S.;Bae,Y.H.In Vivo Evaluation of Doxorubicin-Loaded Polymeric Micelles Targeting Folate Receptors and Early Endosomal p H in Drug-Resistant Ovarian Cancer.Mol.Pharm.2009,6,1353-1362.