Water-Soluble Cationic Polyphosphazenes Grafted with Cyclic Polyamine and Imidazole as an Effective Gene Delivery Vector.

Gene therapy holds immense potential as a future therapeutic strategy for the treatment of numerous genetic diseases which are incurable to date. Nevertheless, safe and efficient gene delivery remains the most challenging aspects of gene therapy. In this study, a series of polyphosphazenes (PPZ) bearing cyclic polyamine and imidazole groups were synthesized and investigated for gene delivery. Agarose gel electrophoresis assays showed that poly(imidazole/1,4,7,10-tetraazyclodocane)phosphazene (Im-PPZ-cyclen) had good binding ability with plasmid DNA (pDNA), yielding positively charged particles with a size around 120-140 nm from a ratio of 10:1 to 5:1 (Im-PPZ-cyclen/pDNA, w/w). The cytotoxicity of Im-PPZ-cyclen assayed by MTT was lower than that of PEI 25 kDa, and was similar to that reported for poly(di-2-dimethylaminoethylamine)phosphazene (poly(di-DMAEA)phosphazene) to some degree. The maximum transfection efficiency of Im-PPZ-cyclen/pDNA complexes against 293 T cells at the ratio of 5:1 (Im-PPZ-cyclen/pDNA, w/w) is close to that of Lipofectamine 2000. The present work may provide a strategy for the design of new cationic polymers with reduced cytotoxicity and be applied to gene delivery as an efficient nonviral vector.

[1]  C. Li,et al.  Chitosan Grafted with Phosphorylcholine and Macrocyclic Polyamine as an Effective Gene Delivery Vector: Preparation, Characterization and In Vitro Transfection. , 2015, Macromolecular bioscience.

[2]  Roberta Cavalli,et al.  Micro- and nanobubbles: a versatile non-viral platform for gene delivery. , 2013, International journal of pharmaceutics.

[3]  M. Kamihira,et al.  Development of hybrid viral vectors for gene therapy. , 2013, Biotechnology advances.

[4]  N. Zhang,et al.  The targeted co-delivery of DNA and doxorubicin to tumor cells via multifunctional PEI-PEG based nanoparticles. , 2013, Biomaterials.

[5]  R. Samulski,et al.  Viral vectors for gene delivery to the central nervous system , 2012, Neurobiology of Disease.

[6]  Bingqing Wang,et al.  Effects of hydrophobic and hydrophilic modifications on gene delivery of amphiphilic chitosan based nanocarriers. , 2011, Biomaterials.

[7]  Yuyang Jiang,et al.  Chitosan grafted with macrocyclic polyamines on C-2 and C-6 positions as nonviral gene vectors: preparation, characterization, and in vitro transfection studies. , 2011, Biomacromolecules.

[8]  Xiang Gao,et al.  Nonviral Gene Delivery: Principle, Limitations, and Recent Progress , 2009, The AAPS Journal.

[9]  M. Ramezani,et al.  Gene transfer efficiency of high primary amine content, hydrophobic, alkyl-oligoamine derivatives of polyethylenimine. , 2009, Biomaterials.

[10]  Bijan Ranjbar,et al.  Circular Dichroism Techniques: Biomolecular and Nanostructural Analyses‐ A Review , 2009, Chemical biology & drug design.

[11]  Meredith A Mintzer,et al.  Nonviral vectors for gene delivery. , 2009, Chemical reviews.

[12]  V. Vlassov,et al.  Transfection Efficiency of 25-kDa PEI–Cholesterol Conjugates with Different Levels of Modification , 2009, Journal of biomaterials science. Polymer edition.

[13]  M. Ogris,et al.  Oligoethylenimine-grafted polypropylenimine dendrimers as degradable and biocompatible synthetic vectors for gene delivery. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[14]  E. Junquera,et al.  A physicochemical characterization of the interaction between DC-Chol/DOPE cationic liposomes and DNA. , 2008, The journal of physical chemistry. B.

[15]  Guoyuan Lu,et al.  DNA binding and cleaving activity of the new cleft molecule N,N'-Bis(guanidinoethyl)-2,6-pyridinedicarboxamide in the absence or in the presence of copper(II). , 2008, Bioconjugate chemistry.

[16]  Yaping Li,et al.  Poly(imidazole/DMAEA)phosphazene/DNA self-assembled nanoparticles for gene delivery: synthesis and in vitro transfection. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[17]  J. Nah,et al.  Chemical modification of chitosan as a gene carrier in vitro and in vivo , 2007 .

[18]  H. Lee,et al.  A tetra(L-lysine)-grafted poly(organophosphazene) for gene delivery. , 2007, Bioorganic & medicinal chemistry letters.

[19]  Mark E. Davis,et al.  Imidazole groups on a linear, cyclodextrin-containing polycation produce enhanced gene delivery via multiple processes. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[20]  Shubiao Zhang,et al.  Toxicity of cationic lipids and cationic polymers in gene delivery. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[21]  W. Hennink,et al.  In vivo tumor transfection mediated by polyplexes based on biodegradable poly(DMAEA)-phosphazene. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[22]  K. Yoshikawa,et al.  The effect of backbone structure on polycation comb-type copolymer/DNA interactions and the molecular assembly of DNA. , 2005, Biomaterials.

[23]  Naoki Kanayama,et al.  Lactose-conjugated polyion complex micelles incorporating plasmid DNA as a targetable gene vector system: their preparation and gene transfecting efficiency against cultured HepG2 cells. , 2004, Journal of Controlled Release.

[24]  F. J. García Alonso,et al.  A simplified and convenient laboratory-scale preparation of 14N or 15N high molecular weight poly(dichlorophosphazene) directly from PCl5. , 2003, Chemistry.

[25]  C. van Nostrum,et al.  Water-soluble biodegradable cationic polyphosphazenes for gene delivery. , 2003, Journal of controlled release : official journal of the Controlled Release Society.

[26]  Thomas Kissel,et al.  In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis. , 2003, Biomaterials.

[27]  Jindrich Kopecek,et al.  Prospects for cationic polymers in gene and oligonucleotide therapy against cancer. , 2002, Advanced drug delivery reviews.

[28]  A. Gewirtz,et al.  Nucleic-acid therapeutics: basic principles and recent applications , 2002, Nature Reviews Drug Discovery.

[29]  M. Ogris,et al.  Polylysine-based transfection systems utilizing receptor-mediated delivery. , 1998, Advanced drug delivery reviews.

[30]  K. Kataoka,et al.  Remarkable increase in nuclease resistance of plasmid DNA through supramolecular assembly with poly(ethylene glycol)-poly(L-lysine) block copolymer. , 1998, Journal of pharmaceutical sciences.

[31]  M. Hashida,et al.  In vitro cytotoxicity of macromolecules in different cell culture systems , 1995 .

[32]  J. Behr,et al.  Gene transfer with synthetic cationic amphiphiles: prospects for gene therapy. , 1994, Bioconjugate chemistry.

[33]  B. Baguley,et al.  Quenching of DNA-ethidium fluorescence by amsacrine and other antitumor agents: a possible electron-transfer effect. , 1984, Biochemistry.

[34]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[35]  H. Allcock,et al.  Hydrolysis pathways for aminophosphazenes , 1982 .

[36]  H. Allcock,et al.  Phosphazene High Polymers with Steroidal Side Groups , 1980 .

[37]  J. Lepecq,et al.  A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization. , 1967, Journal of molecular biology.

[38]  J. Nah,et al.  Target gene delivery from targeting ligand conjugated chitosan-PEI copolymer for cancer therapy. , 2016, Carbohydrate polymers.

[39]  E. Wagner,et al.  Pyridylhydrazone-based PEGylation for pH-reversible lipopolyplex shielding. , 2011, Biomaterials.

[40]  P. Midoux,et al.  Histidylated polylysine as DNA vector: elevation of the imidazole protonation and reduced cellular uptake without change in the polyfection efficiency of serum stabilized negative polyplexes. , 2001, Bioconjugate chemistry.

[41]  E. Schacht,et al.  Biodegradable polymers. II. Degradation characteristics of hydrolysis-sensitive poly[(organo)phosphazenes]. , 1992, Biomaterials.

[42]  E. Schacht,et al.  Biodegradable polymers. I. Synthesis of hydrolysis-sensitive poly[(organo)phosphazenes]. , 1992, Biomaterials.

[43]  D. P. Mack,et al.  Synthesis of Poly[(amino acid alkyl ester)phosphazenes] , 1977 .