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Reversibly crosslinked pH-sensitive polymersomes for efficient intracellular protein delivery
e104
Abstracts / Journal of Controlled Release 172 (2013) e98–e124
Reversibly crosslinked pH-sensitive polymersomes for efficient intracellular protein delivery Huanli Sun, Fenghua Meng, Ru Cheng, Chao Deng, Zhiyuan Zhong⁎ Biomedical Polymers Laboratory, Jiangsu Key Laboratory of Advanced Functional Polymer Design and Application, College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou 215123, China E-mail addresses: [email protected] (H. Sun), [email protected] (Z. Zhong). In recent years, polymersomes with large aqueous compartments as well as robust hydrophobic membranes have emerged as one of the most ideal nanocarriers for encapsulation and controlled delivery of therapeutic proteins and peptides [1]. However, polymersomes often encounter issues of low protein loading contents, slow drug release at the site of action and premature release following i.v. administration. Herein, we report on reversibly crosslinked pHsensitive polymersomes (CLPs) based on poly(ethylene glycol)-bpoly(acrylic acid)(thiol)-b-poly(2-(diethyl amino)ethyl methacrylate) (PEG-PAA(SH)-PDEA) triblock copolymers for the efficient loading of proteins under mild conditions as well as rapid intracellular protein release (Scheme 1). By adjusting the pH to 7.8 PEG-PAA(SH)-PDEA readily formed mono-disperse polymersomes (ca. 35 nm), which could facilely be crosslinked by forming disulfide bonds when exposed in the air. The CLPs, while showing remarkable stability, were rapidly dissociated under reductive conditions. These polymersomes could efficiently load proteins. Notably, proteinloaded CLPs demonstrated fast and quantitative release of proteins in the presence of 10 mM GSH at pH 7.4 and 5.0. The cell experiments showed that protein-loaded CLPs accomplished efficient delivery and release of proteins into MCF-7 cells. Importantly, the intracellularly released proteins retained their biological activity. These reversibly crosslinked pH-sensitive polymersomes are promising for intracellular delivery of peptide and protein drugs.
Reference [1] F.H. Meng, Z.Y. Zhong, Polymersomes spanning from nano- to microscales: advanced vehicles for controlled drug delivery and robust vesicles for virus and cell mimicking, J. Phys. Chem. Lett. 2 (2011) 1533–1539.
doi:10.1016/j.jconrel.2013.08.255
Quaternized poly(glycerol methacrylate)s for enhanced pDNA delivery Hui Gaoa, Zhixiang Lianga, Xinshi Wua, Cui Lia, Ying-Wei Yangb, Guolin Wuc, Jianbiao Maa a School of Chemistry and Chemical Engineering, Tianjin University of Technology, Tianjin 300384, China b State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, China c Key Laboratory of Functional Polymer Materials (Ministry of Education), Institute of Polymer Chemistry, Nankai University, Tianjin 300071, China E-mail address: [email protected] (H. Gao). Low transfection efficiency and undesirable cytotoxicity remain the most challenging aspects for cationic polymers as gene carrier. To overcome the disadvantages, various modifications have been made to improve their gene delivery efficacy. Among them, quaternary ammonium salt-functionalized amino groups of cationic polymers displayed promising results. In this study both linear and 8-arm amino poly (glycerol methacrylate)s (PGOHMAs) were synthesized from poly (glycidyl methacrylate)s (PGMA)s via ring opening reactions with methylethylamine (MEA) [1], diethylamine (DEA) and dipropylamine (DPA), respectively. Further modification of tertiary amine groups on amino PGMAs with quaternization reaction using methyl iodide resulted in partially quaternized PGMAs, namely, QPGMAs. QPGMA/pDNA complexes at different nitrogen-to-phosphate (N/P) ratios were then prepared (Fig. 1). In addition, eight-arm star-shaped MEA modified PGMA (S8-MEA) was also used to complex with pDNA as a positive control to compare with QPGMA/pDNA complexes. QPGMA/pDNA complexes are spherical in shape as observed by scanning electron microscopy (SEM), and are smaller in size than S8-MEA/pDNA complexes as suggested by dynamic light scattering (DLS) measurement. Interestingly, the gel electrophoresis and ethidium bromide displacement assay indicated that QPGMA polymers can condense pDNA at lower N/P ratios than S8-MEA. In vitro experiments on Huh-7cells showed that QPGMAs displayed lower cytotoxicity and better transfection efficacy, compared to S8-PGMA. The cellular uptake analysis results revealed that QPGMA enhanced the cellular uptake level of pDNA. Though the endosomal escape efficiency of QS8-MEA was a little bit lower than that of S8-MEA, on the whole, amino PGOHMAs modified by quaternization were proved to be effective systemic gene vectors for gene therapy.
Scheme 1. Illustration of reversibly crosslinked pH-sensitive polymersomes based on PEG-PAA(SH)-PDEA triblock copolymers for triggered intracellular release of proteins.
Keywords: Polymersomes, Reversibly crosslinking, pH-sensitive, Protein delivery Acknowledgments This work was supported by the National Natural Science Foundation of China (NSFC 20974073, 50973078 and 51173126), the Innovative Graduate Research Program of Jiangsu Province (Grant Nos. CXZZ11_0094), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
Fig. 1. Schematic illustration of QPGMA/pDNA complex formation.
Keywords: Quaternization, Poly(glycerol methacrylate)s, pDNA, Transfection efficacy Acknowledgments Financial support from NSFC (21074092) and Program for New Century Excellent Talents in University (NCET-11-1063) is highly acknowledged.
Abstracts / Journal of Controlled Release 172 (2013) e98–e124
Reversibly crosslinked pH-sensitive polymersomes for efficient intracellular protein delivery Huanli Sun, Fenghua Meng, Ru Cheng, Chao Deng, Zhiyuan Zhong⁎ Biomedical Polymers Laboratory, Jiangsu Key Laboratory of Advanced Functional Polymer Design and Application, College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou 215123, China E-mail addresses: [email protected] (H. Sun), [email protected] (Z. Zhong). In recent years, polymersomes with large aqueous compartments as well as robust hydrophobic membranes have emerged as one of the most ideal nanocarriers for encapsulation and controlled delivery of therapeutic proteins and peptides [1]. However, polymersomes often encounter issues of low protein loading contents, slow drug release at the site of action and premature release following i.v. administration. Herein, we report on reversibly crosslinked pHsensitive polymersomes (CLPs) based on poly(ethylene glycol)-bpoly(acrylic acid)(thiol)-b-poly(2-(diethyl amino)ethyl methacrylate) (PEG-PAA(SH)-PDEA) triblock copolymers for the efficient loading of proteins under mild conditions as well as rapid intracellular protein release (Scheme 1). By adjusting the pH to 7.8 PEG-PAA(SH)-PDEA readily formed mono-disperse polymersomes (ca. 35 nm), which could facilely be crosslinked by forming disulfide bonds when exposed in the air. The CLPs, while showing remarkable stability, were rapidly dissociated under reductive conditions. These polymersomes could efficiently load proteins. Notably, proteinloaded CLPs demonstrated fast and quantitative release of proteins in the presence of 10 mM GSH at pH 7.4 and 5.0. The cell experiments showed that protein-loaded CLPs accomplished efficient delivery and release of proteins into MCF-7 cells. Importantly, the intracellularly released proteins retained their biological activity. These reversibly crosslinked pH-sensitive polymersomes are promising for intracellular delivery of peptide and protein drugs.
Reference [1] F.H. Meng, Z.Y. Zhong, Polymersomes spanning from nano- to microscales: advanced vehicles for controlled drug delivery and robust vesicles for virus and cell mimicking, J. Phys. Chem. Lett. 2 (2011) 1533–1539.
doi:10.1016/j.jconrel.2013.08.255
Quaternized poly(glycerol methacrylate)s for enhanced pDNA delivery Hui Gaoa, Zhixiang Lianga, Xinshi Wua, Cui Lia, Ying-Wei Yangb, Guolin Wuc, Jianbiao Maa a School of Chemistry and Chemical Engineering, Tianjin University of Technology, Tianjin 300384, China b State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, China c Key Laboratory of Functional Polymer Materials (Ministry of Education), Institute of Polymer Chemistry, Nankai University, Tianjin 300071, China E-mail address: [email protected] (H. Gao). Low transfection efficiency and undesirable cytotoxicity remain the most challenging aspects for cationic polymers as gene carrier. To overcome the disadvantages, various modifications have been made to improve their gene delivery efficacy. Among them, quaternary ammonium salt-functionalized amino groups of cationic polymers displayed promising results. In this study both linear and 8-arm amino poly (glycerol methacrylate)s (PGOHMAs) were synthesized from poly (glycidyl methacrylate)s (PGMA)s via ring opening reactions with methylethylamine (MEA) [1], diethylamine (DEA) and dipropylamine (DPA), respectively. Further modification of tertiary amine groups on amino PGMAs with quaternization reaction using methyl iodide resulted in partially quaternized PGMAs, namely, QPGMAs. QPGMA/pDNA complexes at different nitrogen-to-phosphate (N/P) ratios were then prepared (Fig. 1). In addition, eight-arm star-shaped MEA modified PGMA (S8-MEA) was also used to complex with pDNA as a positive control to compare with QPGMA/pDNA complexes. QPGMA/pDNA complexes are spherical in shape as observed by scanning electron microscopy (SEM), and are smaller in size than S8-MEA/pDNA complexes as suggested by dynamic light scattering (DLS) measurement. Interestingly, the gel electrophoresis and ethidium bromide displacement assay indicated that QPGMA polymers can condense pDNA at lower N/P ratios than S8-MEA. In vitro experiments on Huh-7cells showed that QPGMAs displayed lower cytotoxicity and better transfection efficacy, compared to S8-PGMA. The cellular uptake analysis results revealed that QPGMA enhanced the cellular uptake level of pDNA. Though the endosomal escape efficiency of QS8-MEA was a little bit lower than that of S8-MEA, on the whole, amino PGOHMAs modified by quaternization were proved to be effective systemic gene vectors for gene therapy.
Scheme 1. Illustration of reversibly crosslinked pH-sensitive polymersomes based on PEG-PAA(SH)-PDEA triblock copolymers for triggered intracellular release of proteins.
Keywords: Polymersomes, Reversibly crosslinking, pH-sensitive, Protein delivery Acknowledgments This work was supported by the National Natural Science Foundation of China (NSFC 20974073, 50973078 and 51173126), the Innovative Graduate Research Program of Jiangsu Province (Grant Nos. CXZZ11_0094), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
Fig. 1. Schematic illustration of QPGMA/pDNA complex formation.
Keywords: Quaternization, Poly(glycerol methacrylate)s, pDNA, Transfection efficacy Acknowledgments Financial support from NSFC (21074092) and Program for New Century Excellent Talents in University (NCET-11-1063) is highly acknowledged.