Synthesis and characterization of novel water-soluble polyamide based on spermine and aspartic acid as a potential gene delivery vehicle

We developed a novel and convenient method for the synthesis of a potentially safe non-viral gene delivery vehi- cle based on the cationic block copolymer of spermine and aspartic acid (ASSP) and coupled it with polyethylene glycol (PEG). The copolymer ASSP was prepared by direct polycondensation in the ionic liquid, butylmethylimidazolium hexa- fluorophosphate, using triphenyl phosphite as the condensing agent under mild reaction conditions. The highly hydropho- bic ASSP was transformed into a water soluble hydrophilic micelle by coupling ASSP with polyethylene glycol (PEG) using the same ionic liquid and 1,1-carbonyl diimidazole as the condensing agent without harsh conditions. The polyca- tionic ASSP-PEG was then used to condense calf thymus and plasmid deoxyribonuclceic acids (DNAs) in Tris-HCl buffer (pH 7.4) to get a series of block ionomer complexes with various charge ratios. The physicochemical properties of the copolymer micelle and the DNA polyplexes were studied using fourier transform-infrared (FTIR), nuclear magnetic reso- nance (NMR) and circular dichroism (CD) spectroscopy, matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), differential scanning calorimetry (DSC), transmission electron microscopy (TEM) and particle size measurements. It was observed that the DNA was condensed to compact particles by its interaction with the copolymer. Since DNA condensation to nano/micrometer sized particles is essential for gene delivery, our results indicate a potential use of the copolymer for gene delivery applications.

[1]  F. Sánchez-Jiménez,et al.  Study by optical spectroscopy and molecular dynamics of the interaction of acridine-spermine conjugate with DNA. , 2008, Biophysical chemistry.

[2]  Tae Gwan Park,et al.  Molecular design of functional polymers for gene therapy , 2007 .

[3]  Eun Seong Lee,et al.  Polymeric nanovehicles for anticancer drugs with triggering release mechanisms , 2007 .

[4]  C. Wu,et al.  A novel potential cationic polymeric gene vector containing hydroxy groups and all grades of amino groups , 2007 .

[5]  D. Putnam,et al.  Polymer systems for gene delivery - Past, present, and future , 2007 .

[6]  G. Pasut,et al.  Polymer-drug conjugation, recent achievements and general strategies , 2007 .

[7]  J. Nah,et al.  A Poly(β-Amino Ester) of Spermine and Poly(ethylene Glycol) Diacrylate as a Gene Carrier , 2007 .

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

[9]  Jun Li,et al.  Cationic star polymers consisting of alpha-cyclodextrin core and oligoethylenimine arms as nonviral gene delivery vectors. , 2007, Biomaterials.

[10]  C. Alexander,et al.  Varying polymer architecture to deliver drugs , 2007, The AAPS Journal.

[11]  William H. Heath,et al.  Charged Polymers via Controlled Radical Polymerization and their Implications for Gene Delivery , 2007 .

[12]  Y. Barenholz,et al.  Characterization and in vivo performance of dextran-spermine polyplexes and DOTAP/cholesterol lipoplexes administered locally and systemically. , 2007, Biomaterials.

[13]  G. Damonte,et al.  The Peptide Nucleic Acid Targeted to a Regulatory Sequence of the Translocated c-myc Oncogene in Burkitt's Lymphoma Lacks Immunogenicity: Follow-Up Characterization of PNAEμ-NLS , 2007 .

[14]  K. Lee.  Chitosan and its derivatives for gene delivery , 2007 .

[15]  N. Kasyanenko,et al.  DNA interaction with synthetic polymers in solution , 2007 .

[16]  J. Nah,et al.  Chitosan-graft-polyethylenimine as a gene carrier. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[17]  Y. Tabata,et al.  Expression profile of plasmid DNA obtained using spermine derivatives of pullulan with different molecular weights , 2007, Journal of biomaterials science. Polymer edition.

[18]  David Putnam,et al.  Polymers for gene delivery across length scales , 2006, Nature materials.

[19]  K. Leong,et al.  Chitosan‐g‐PEG/DNA complexes deliver gene to the rat liver via intrabiliary and intraportal infusions , 2006, The journal of gene medicine.

[20]  Lonnie D Shea,et al.  Design of modular non-viral gene therapy vectors. , 2006, Biomaterials.

[21]  S. B. Tiwari,et al.  Long-Circulating Polymeric Nanovectors for Tumor-Selective Gene Delivery , 2005, Technology in cancer research & treatment.

[22]  Daniel W. Pack,et al.  Design and development of polymers for gene delivery , 2005, Nature Reviews Drug Discovery.

[23]  T. Thomas,et al.  Nanotechnology in Nonviral Gene Delivery , 2005 .

[24]  D. Burgess,et al.  DNA-based therapeutics and DNA delivery systems: A comprehensive review , 2005, The AAPS Journal.

[25]  H. Lipps,et al.  Towards safe, non-viral therapeutic gene expression in humans , 2005, Nature Reviews Genetics.

[26]  T. Thomas,et al.  Ionic, structural, and temperature effects on DNA nanoparticles formed by natural and synthetic polyamines. , 2005, Biomacromolecules.

[27]  C. Alexander,et al.  Synthetic polymers as drugs and therapeutics , 2005 .

[28]  Y. Barenholz,et al.  Polymers for DNA Delivery , 2005, Molecules.

[29]  M. Linial,et al.  Hydrophobized dextran-spermine conjugate as potential vector for in vitro gene transfection. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[30]  T. Thomas,et al.  Formation of DNA nanoparticles in the presence of novel polyamine analogues: a laser light scattering and atomic force microscopic study. , 2004, Nucleic acids research.

[31]  T. Thomas,et al.  DNA nanoparticles and development of DNA delivery vehicles for gene therapy. , 2002, Biochemistry.

[32]  T. Thomas,et al.  Polyamine structural effects on the induction and stabilization of liquid crystalline DNA: potential applications to DNA packaging, gene therapy and polyamine therapeutics. , 2002, Nucleic acids research.

[33]  E. I. Lozinskaya,et al.  Ionic Liquids as Novel Reaction Media for the Synthesis of Condensation Polymers , 2002 .

[34]  R. Jerome,et al.  Morphology of Water-Soluble Interpolyelectrolyte Complexes Formed by Poly(2-vinylpyridinium)-block-poly(ethylene oxide) Diblocks and Poly(4-styrenesulfonate) Polyanions , 2001 .

[35]  A. Kabanov,et al.  Recognition of DNA Topology in Reactions between Plasmid DNA and Cationic Copolymers , 2000 .

[36]  M. A. Cohen Stuart,et al.  Formation of Micelles with Complex Coacervate Cores , 1998 .

[37]  A. Kabanov,et al.  Self-assembly of polyamine-poly(ethylene glycol) copolymers with phosphorothioate oligonucleotides. , 1998, Bioconjugate chemistry.

[38]  Cheng,et al.  Airway delivery of cationic lipid: DNA complexes for cystic fibrosis. , 1998, Advanced drug delivery reviews.

[39]  Atsushi Harada,et al.  Spontaneous Formation of Polyion Complex Micelles with Narrow Distribution from Antisense Oligonucleotide and Cationic Block Copolymer in Physiological Saline , 1996 .

[40]  S. Zimmerman,et al.  Macromolecular crowding and the mandatory condensation of DNA in bacteria , 1996, FEBS letters.

[41]  S. V. Vinogradov,et al.  Water-soluble block polycations as carriers for oligonucleotide delivery. , 1995, Bioconjugate chemistry.

[42]  D. Scherman,et al.  A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[43]  G. Filipcsei.  Smart polymers for biomedical application and environmental protection , 2008 .

[44]  J. Rossi,et al.  Gene transfer : delivery and expression of DNA and RNA : a laboratory manual , 2007 .

[45]  金谷 勲.  Efficient gene transfer by pullulan-spermine occurs through both clathrin- and raft/caveolae-dependent mechanisms , 2007 .

[46]  M. Glodde,et al.  Physiochemical properties of low and high molecular weight poly(ethylene glycol)-grafted poly(ethylene imine) copolymers and their complexes with oligonucleotides. , 2006, Biomacromolecules.

[47]  Leaf Huang,et al.  Recent Advances in Non-viral Gene Delivery. , 2005, Advances in genetics.

[48]  Y. Tabata,et al.  Dextran–spermine polycation: an efficient nonviral vector for in vitro and in vivo gene transfection , 2004, Gene Therapy.

[49]  Robin D. Rogers,et al.  Room temperature ionic liquids as novel media for ‘clean’ liquid–liquid extraction , 1998 .

[50]  A. Rolland.  From genes to gene medicines: recent advances in nonviral gene delivery. , 1998, Critical reviews in therapeutic drug carrier systems.

[51]  V. Bloomfield.  DNA condensation by multivalent cations. , 1997, Biopolymers.

[52]  Atsushi Harada,et al.  Formation of Polyion Complex Micelles in an Aqueous Milieu from a Pair of Oppositely-Charged Block Copolymers with Poly(ethylene glycol) Segments , 1995 .