pH响应型苯硼酸酯连接嵌段聚合物的合成及药物控释
|
摘要
以含苯硼酸酯(PBE)的聚乙二醇单甲醚(mPEG)大分子(mPEG-PBE-OH)为引发剂,引发ε-己内酯(ε-CL)开环聚合,制备了以硼酸酯结构连接的pH敏感两亲性聚合物(mPEG-PBEPCL)。然后,使该聚合物在水相环境中自组装形成"核-壳"结构纳米胶束,并将阿霉素(DOX)负载在胶束内核中,形成载药胶束(DOX@mPEG-PBE-PCL)。通过核磁共振氢谱(~1 H-NMR)、红外光谱(FT-IR)和凝胶渗透色谱(GPC)对聚合物结构进行了表征,通过透射电镜(TEM)和动态光散射(DLS)等对胶束的形貌和粒径进行了表征,通过紫外吸收光谱分析了胶束载药量和载药效率,并对胶束的pH敏感释药性能与体外细胞毒性进行了验证。结果表明:聚合物自组装形成粒径约127nm的球形胶束,对DOX具有较高的负载能力;聚合物具有良好的pH响应性和生物相容性,DOX@mPEG-PBE-PCL能在肿瘤细胞弱酸性环境中释放DOX,有效递送至细胞核;与游离的DOX·HCl相比,DOX@mPEG-PBE-PCL对鼠源黑色素瘤B16F10细胞具有相近的抗肿瘤活性。
A pH-sensitive amphiphilic polymer Poly(ethylene glycol)-phenylborate-poly(ε-caprolactone)(mPEG-PBE-PCL)linked by a borate structure was prepared by ring-opening polymerization ofε-caprolactone(ε-CL)initiated by a benzene-borate-containing polyethylene glycol macromolecule(mPEGPBE-OH).The polymer was capable of self-assembling into a "core-shell"nanomicelles structure in aqueous environment,which can load doxorubicin(DOX)in their cores.The structure of polymer was characterized by ~1 H-NMR,IR spectra and gel permeation chromatography(GPC).The morphology and particle size of the prepared micelles were determined by transmission electron microscopy(TEM)and dynamic light scattering(DLS).The drug loading and loading efficiency were analyzed by ultraviolet absorption detection.The investigations on pH-sensitive release properties of the micelles and the in vitro cytotoxicity were carried out as well.The results showed that the polymer micelles had a low critical micelle concentration,showed spherical micelles with a diameter of about 127 nm,and had a high loading capacity for DOX.Furthermore,the polymer presented good pH responsiveness and biocompatibility.The loaded DOX was released in weak acidic environment of tumor cells and was delivered effectively to nucleus.DOX@mPEG-PBE-PCL had a similar antitumor activity with free DOX·HCl against murine melanoma B16 F10 cells.Therefore,such smart micelles containing pH-responsive boronate bonds present great potential in efficient intracellular delivery of antitumor drugs.
A pH-sensitive amphiphilic polymer Poly(ethylene glycol)-phenylborate-poly(ε-caprolactone)(mPEG-PBE-PCL)linked by a borate structure was prepared by ring-opening polymerization ofε-caprolactone(ε-CL)initiated by a benzene-borate-containing polyethylene glycol macromolecule(mPEGPBE-OH).The polymer was capable of self-assembling into a "core-shell"nanomicelles structure in aqueous environment,which can load doxorubicin(DOX)in their cores.The structure of polymer was characterized by ~1 H-NMR,IR spectra and gel permeation chromatography(GPC).The morphology and particle size of the prepared micelles were determined by transmission electron microscopy(TEM)and dynamic light scattering(DLS).The drug loading and loading efficiency were analyzed by ultraviolet absorption detection.The investigations on pH-sensitive release properties of the micelles and the in vitro cytotoxicity were carried out as well.The results showed that the polymer micelles had a low critical micelle concentration,showed spherical micelles with a diameter of about 127 nm,and had a high loading capacity for DOX.Furthermore,the polymer presented good pH responsiveness and biocompatibility.The loaded DOX was released in weak acidic environment of tumor cells and was delivered effectively to nucleus.DOX@mPEG-PBE-PCL had a similar antitumor activity with free DOX·HCl against murine melanoma B16 F10 cells.Therefore,such smart micelles containing pH-responsive boronate bonds present great potential in efficient intracellular delivery of antitumor drugs.
引文
[1] ZHAO D,XU J Q,YI X Q,et al.pH-activated targeting drug delivery system based on the selective binding of phenylboronic acid[J].ACS Applied Materials&Interfaces,2016,8(23):14845-14854.
[2] BURGESS D J,DOLES J,ZENDER L,et al.Topoisomerase levels determine chemotherapy response in vitro and in vivo[J].Proceedings of the National Academy of Sciences,2008,105(26):9053-9058.
[3] WANG J,XU W,LI S,et al.Polylactide-cholesterol stereocomplex micelle encapsulating chemotherapeutic agent for improved antitumor efficacy and safety[J].Journal of Biomedical Nanotechnology,2018,14(12):2102-2113.
[4] MENG F,ZHONG Y,CHENG R,et al.pH-sensitive polymeric nanoparticles for tumor-targeting doxorubicin delivery:Concept and recent advances[J].Nanomedicine,2014,9(3):487-499.
[5]王曦阳,王灏,郭和泽,等.兼具MRI显影增强及pH敏感释药性能的载药Fe3O4纳米颗粒[J].功能高分子学报,2016,29(2):193-199.
[6] PRABHAKAR U,MAEDA H,JAIN R K,et al.Challenges and key considerations of the enhanced permeability and retention effect for nanomedicine drug delivery in oncology[J].Cancer Research,2013,73(8):2412-2417.
[7] MENG F,ZHONG Y,CHENG R,et al.pH-sensitive polymeric nanoparticles for tumor-targeting doxorubicin delivery:Concept and recent advances[J].Nanomedicine,2014,9(3):487-499.
[8] LIU J,HUANG Y,KUMAR A,et al.pH-sensitive nano-systems for drug delivery in cancer therapy[J].Biotechnology Advances,2014,32(4):693-710.
[9] XU W,DING J,CHEN X.Reduction-responsive polypeptide micelles for intracellular delivery of antineoplastic agent[J].Biomacromolecules,2017,18(10):3291-3301.
[10] HE X,DING M,LI J,et al.Biodegradable multiblock polyurethane micelles with tunable reduction-sensitivity for on-demand intracellular drug delivery[J].Rsc Advances,2014,4(47):24736-24746.
[11] MANNA U,PATIC S.Glucose-triggered drug delivery from borate mediated layer-by-layer self-assembly[J].ACS Applied Materials&Interfaces,2010,2(5):1521-1527.
[12] KANNAGI R,SAKUMA K,MIYAZAKI K,et al.Altered expression of glycan genes in cancers induced by epigenetic silencing and tumor hypoxia:Clues in the ongoing search for new tumor markers[J].Cancer Science,2010,101(3):586-593.
[13] DOHI T,HASHIGUCH M,YAMAMOTO S,et al.Fucosyltransferase-producing sialyl Le(a)and sialyl Le(x)carbohydrate antigen in benign and malignant gastrointestinal mucosa[J].Cancer,2015,73(6):1552-1561.
[14] ULBRICH K,HOLA K,SUBR V,et al.Targeted drug delivery with polymers and magnetic nanoparticles:Covalent and noncovalent approaches,release control,and clinical studies[J].Chemical Reviews,2016,116(9):5338-5431.
[15] MACEWAN S R,CALLAHAN D J,CHILKOTI A.Stimulus-responsive macromolecules and nanoparticles for cancer drug delivery[J].Nanomedicine,2010,5(5):793-806.
[16] TORCHILIN V.Tumor delivery of macromolecular drugs based on the EPR effect[J].Advanced Drug Delivery Reviews,2011,63(3):131-135.
[17] ANZAI J.Recent progress in electrochemical biosensors based on phenylboronic acid and derivatives[J].Materials Science and Engineering:C,2016,67:737-746.
[18] YAO Y,ZHAO L,YANG J,et al.Glucose-responsive vehicles containing phenylborate ester for controlled insulin release at neutral pH[J].Biomacromolecules,2012,13(6):1837-1844.
[19] ZHANG C,LOSEGO M D,BRAUN P V.Hydrogel-based glucose sensors:Effects of phenylboronic acid chemical structure on response[J].Chemistry of Materials,2013,25(15):3239-3250.
[20] HWANG S J,FERNANDEZ C,AMOUREUX J P,et al.Structural study of x Na2S(+1-x)B2S3glasses and polycrystals by multiple-quantum MAS NMR of 11 B and 23 Na[J].Journal of the American Chemical Society,1998,120(29):7337-7346.
[21] MATSUMOTO A,IKEDA S,HARADA A,et al.Glucose-responsive polymer bearing a novel phenylborate derivative as a glucose-sensing moiety operating at physiological pH conditions[J].Biomacromolecules,2003,4(5):1410-1416.
[22] MULLA H R,AGARD N J,BASU A.3-Methoxycarbonyl-5-nitrophenyl boronic acid:High affinity diol recognition at neutral Ph[J].Bioorganic&Medicinal Chemistry Letters,2004,14(1):25-27.
[23] MATSUMOTO A,ISHII T,NISHIDA J,et al.A synthetic approach toward a self-regulated insulin delivery system[J].Angewandte Chemie International Edition,2012,51(9):2124-2128.
[24] KIM K T,CORNELISSEN J J L M,NOLTE R J M,et al.Polymeric monosaccharide receptors responsive at neutral pH[J].Journal of the American Chemical Society,2009,131(39):13908-13909.
[25] ZHAO L,HUANG Q,LIU Y,et al.Boronic acid as glucose-sensitive agent regulates drug delivery for diabetes treatment[J].Materials,2017,10(2):170-184.
[26] BROOKS W L A,SUMERLIN B S.Synthesis and applications of boronic acid-containing polymers:From materials to medicine[J].Chemical Reviews,2015,116(3):1375-1397.
[27] LIU H,LI Y,SUN K,et al.Dual-responsive surfaces modified with phenylboronic acid-containing polymer brush to reversibly capture and release cancer cells[J].Journal of the American Chemical Society,2013,135(20):7603-7609.
[28] CHEN J,DING J,ZHANG Y,et al.Polyion complex micelles with gradient pH-sensitivity for adjustable intracellular drug delivery[J].Polymer Chemistry,2015,6(3):397-405.
[29] WANG M,LIU T, HAN L,et al.Functionalized o-carboxymethyl-chitosan/polyethylenimine based novel dual pH-responsive nanocarriers for controlled co-delivery of DOX and genes[J].Polymer Chemistry,2015,6(17):3324-3335.
[30] ARNOLD A E,CZUPIEL P,SHOICHET M.Engineered polymeric nanoparticles to guide the cellular internalization and trafficking of small interfering ribonucleic acids[J].Journal of Controlled Release,2017,259(8):3-15.
[31] CASEY J R,GRINSTEIN S,ORLOWSKI J.Sensors and regulators of intracellular pH[J].Nature Reviews Molecular Cell Biology,2010,11(1):50-61.
[32] KIM J S,CHOI D K,SHIN J Y,et al.Endosomal acidic pH-induced conformational changes of a cytosol-penetrating antibody mediate endosomal escape[J].Journal of Controlled Release,2016,235(8):165-175.
[33] PARK K.To PEGylate or not to PEGylate,that is not the question[J].Journal of Controlled Release:Official Journal of the Controlled Release Society,2010,142(2):147-148.
[34] WU H,ZHU L,TORCHILIN V P.pH-sensitive poly(histidine)-PEG/DSPE-PEG co-polymer micelles for cytosolic drug delivery[J].Biomaterials,2013,34(4):1213-1222.
[35] IBUSUKI S,FUJII Y,IWAMOTO Y,et al.Tissue-engineered cartilage using an injectable and in situ gelable thermoresponsive gelatin:Fabrication and in vitro performance[J].Tissue Engineering,2003,9(2):371-384.
[36] JIA H,ZHU J,WANG X,et al.A boronate-linked linear-hyperbranched polymeric nanovehicle for pH-dependent tumor-targeted drug delivery[J].Biomaterials,2014,35(19):5240-5249.
[37] ZHU Y,ZHANG J,MENG F,et al.cRGD-functionalized reduction-sensitive shell-sheddable biodegradable micelles mediate enhanced doxorubicin delivery to human glioma xenografts in vivo[J].Journal of Controlled Release,2016,233(7):29-38.
[38] SALIC A,MITCHISON T J.A chemical method for fast and sensitive detection of DNA synthesis in vivo[J].Proc Natl Acad Sci USA,2008,105(7):2415-2420.
[39] YAMAKOSHI H,DODO K,OKADA M,et al.Imaging of EdU,an alkyne-tagged cell proliferation probe,by Raman microscopy[J].Journal of the American Chemical Society,2011,133(16):6102-6105.
[2] BURGESS D J,DOLES J,ZENDER L,et al.Topoisomerase levels determine chemotherapy response in vitro and in vivo[J].Proceedings of the National Academy of Sciences,2008,105(26):9053-9058.
[3] WANG J,XU W,LI S,et al.Polylactide-cholesterol stereocomplex micelle encapsulating chemotherapeutic agent for improved antitumor efficacy and safety[J].Journal of Biomedical Nanotechnology,2018,14(12):2102-2113.
[4] MENG F,ZHONG Y,CHENG R,et al.pH-sensitive polymeric nanoparticles for tumor-targeting doxorubicin delivery:Concept and recent advances[J].Nanomedicine,2014,9(3):487-499.
[5]王曦阳,王灏,郭和泽,等.兼具MRI显影增强及pH敏感释药性能的载药Fe3O4纳米颗粒[J].功能高分子学报,2016,29(2):193-199.
[6] PRABHAKAR U,MAEDA H,JAIN R K,et al.Challenges and key considerations of the enhanced permeability and retention effect for nanomedicine drug delivery in oncology[J].Cancer Research,2013,73(8):2412-2417.
[7] MENG F,ZHONG Y,CHENG R,et al.pH-sensitive polymeric nanoparticles for tumor-targeting doxorubicin delivery:Concept and recent advances[J].Nanomedicine,2014,9(3):487-499.
[8] LIU J,HUANG Y,KUMAR A,et al.pH-sensitive nano-systems for drug delivery in cancer therapy[J].Biotechnology Advances,2014,32(4):693-710.
[9] XU W,DING J,CHEN X.Reduction-responsive polypeptide micelles for intracellular delivery of antineoplastic agent[J].Biomacromolecules,2017,18(10):3291-3301.
[10] HE X,DING M,LI J,et al.Biodegradable multiblock polyurethane micelles with tunable reduction-sensitivity for on-demand intracellular drug delivery[J].Rsc Advances,2014,4(47):24736-24746.
[11] MANNA U,PATIC S.Glucose-triggered drug delivery from borate mediated layer-by-layer self-assembly[J].ACS Applied Materials&Interfaces,2010,2(5):1521-1527.
[12] KANNAGI R,SAKUMA K,MIYAZAKI K,et al.Altered expression of glycan genes in cancers induced by epigenetic silencing and tumor hypoxia:Clues in the ongoing search for new tumor markers[J].Cancer Science,2010,101(3):586-593.
[13] DOHI T,HASHIGUCH M,YAMAMOTO S,et al.Fucosyltransferase-producing sialyl Le(a)and sialyl Le(x)carbohydrate antigen in benign and malignant gastrointestinal mucosa[J].Cancer,2015,73(6):1552-1561.
[14] ULBRICH K,HOLA K,SUBR V,et al.Targeted drug delivery with polymers and magnetic nanoparticles:Covalent and noncovalent approaches,release control,and clinical studies[J].Chemical Reviews,2016,116(9):5338-5431.
[15] MACEWAN S R,CALLAHAN D J,CHILKOTI A.Stimulus-responsive macromolecules and nanoparticles for cancer drug delivery[J].Nanomedicine,2010,5(5):793-806.
[16] TORCHILIN V.Tumor delivery of macromolecular drugs based on the EPR effect[J].Advanced Drug Delivery Reviews,2011,63(3):131-135.
[17] ANZAI J.Recent progress in electrochemical biosensors based on phenylboronic acid and derivatives[J].Materials Science and Engineering:C,2016,67:737-746.
[18] YAO Y,ZHAO L,YANG J,et al.Glucose-responsive vehicles containing phenylborate ester for controlled insulin release at neutral pH[J].Biomacromolecules,2012,13(6):1837-1844.
[19] ZHANG C,LOSEGO M D,BRAUN P V.Hydrogel-based glucose sensors:Effects of phenylboronic acid chemical structure on response[J].Chemistry of Materials,2013,25(15):3239-3250.
[20] HWANG S J,FERNANDEZ C,AMOUREUX J P,et al.Structural study of x Na2S(+1-x)B2S3glasses and polycrystals by multiple-quantum MAS NMR of 11 B and 23 Na[J].Journal of the American Chemical Society,1998,120(29):7337-7346.
[21] MATSUMOTO A,IKEDA S,HARADA A,et al.Glucose-responsive polymer bearing a novel phenylborate derivative as a glucose-sensing moiety operating at physiological pH conditions[J].Biomacromolecules,2003,4(5):1410-1416.
[22] MULLA H R,AGARD N J,BASU A.3-Methoxycarbonyl-5-nitrophenyl boronic acid:High affinity diol recognition at neutral Ph[J].Bioorganic&Medicinal Chemistry Letters,2004,14(1):25-27.
[23] MATSUMOTO A,ISHII T,NISHIDA J,et al.A synthetic approach toward a self-regulated insulin delivery system[J].Angewandte Chemie International Edition,2012,51(9):2124-2128.
[24] KIM K T,CORNELISSEN J J L M,NOLTE R J M,et al.Polymeric monosaccharide receptors responsive at neutral pH[J].Journal of the American Chemical Society,2009,131(39):13908-13909.
[25] ZHAO L,HUANG Q,LIU Y,et al.Boronic acid as glucose-sensitive agent regulates drug delivery for diabetes treatment[J].Materials,2017,10(2):170-184.
[26] BROOKS W L A,SUMERLIN B S.Synthesis and applications of boronic acid-containing polymers:From materials to medicine[J].Chemical Reviews,2015,116(3):1375-1397.
[27] LIU H,LI Y,SUN K,et al.Dual-responsive surfaces modified with phenylboronic acid-containing polymer brush to reversibly capture and release cancer cells[J].Journal of the American Chemical Society,2013,135(20):7603-7609.
[28] CHEN J,DING J,ZHANG Y,et al.Polyion complex micelles with gradient pH-sensitivity for adjustable intracellular drug delivery[J].Polymer Chemistry,2015,6(3):397-405.
[29] WANG M,LIU T, HAN L,et al.Functionalized o-carboxymethyl-chitosan/polyethylenimine based novel dual pH-responsive nanocarriers for controlled co-delivery of DOX and genes[J].Polymer Chemistry,2015,6(17):3324-3335.
[30] ARNOLD A E,CZUPIEL P,SHOICHET M.Engineered polymeric nanoparticles to guide the cellular internalization and trafficking of small interfering ribonucleic acids[J].Journal of Controlled Release,2017,259(8):3-15.
[31] CASEY J R,GRINSTEIN S,ORLOWSKI J.Sensors and regulators of intracellular pH[J].Nature Reviews Molecular Cell Biology,2010,11(1):50-61.
[32] KIM J S,CHOI D K,SHIN J Y,et al.Endosomal acidic pH-induced conformational changes of a cytosol-penetrating antibody mediate endosomal escape[J].Journal of Controlled Release,2016,235(8):165-175.
[33] PARK K.To PEGylate or not to PEGylate,that is not the question[J].Journal of Controlled Release:Official Journal of the Controlled Release Society,2010,142(2):147-148.
[34] WU H,ZHU L,TORCHILIN V P.pH-sensitive poly(histidine)-PEG/DSPE-PEG co-polymer micelles for cytosolic drug delivery[J].Biomaterials,2013,34(4):1213-1222.
[35] IBUSUKI S,FUJII Y,IWAMOTO Y,et al.Tissue-engineered cartilage using an injectable and in situ gelable thermoresponsive gelatin:Fabrication and in vitro performance[J].Tissue Engineering,2003,9(2):371-384.
[36] JIA H,ZHU J,WANG X,et al.A boronate-linked linear-hyperbranched polymeric nanovehicle for pH-dependent tumor-targeted drug delivery[J].Biomaterials,2014,35(19):5240-5249.
[37] ZHU Y,ZHANG J,MENG F,et al.cRGD-functionalized reduction-sensitive shell-sheddable biodegradable micelles mediate enhanced doxorubicin delivery to human glioma xenografts in vivo[J].Journal of Controlled Release,2016,233(7):29-38.
[38] SALIC A,MITCHISON T J.A chemical method for fast and sensitive detection of DNA synthesis in vivo[J].Proc Natl Acad Sci USA,2008,105(7):2415-2420.
[39] YAMAKOSHI H,DODO K,OKADA M,et al.Imaging of EdU,an alkyne-tagged cell proliferation probe,by Raman microscopy[J].Journal of the American Chemical Society,2011,133(16):6102-6105.