title: Architecturally induced multiresponsive vesicles from well-defined polypeptides: formation of gene vehicles.
abstract: A series of novel, partially labeled amphiphilic triblock copolypeptides, PLL-b-PBLG-d7-b-PLL, has been synthesized, where PLL and PBLG-d7 are poly(L-lysine hydrochloride) and poly(gamma-benzyl-d7-L-glutamate), respectively. The synthetic approach involved the sequential ring-opening polymerization (ROP) of gamma-benzyl-L-glutamate and epsilon-Boc-L-lysine N-carboxy anhydrides by a diamino initiator using high-vacuum techniques, followed by the selective deprotection of the Boc groups. Combined characterization results showed that the copolypeptides exhibit high degrees of molecular and compositional homogeneity. The synthesized copolypeptides had similar molecular weights, while the composition of the middle block ranged between 19 and 74% with respect to the monomeric units. Due to the macromolecular architecture of the copolypeptide and the rigid nature of the middle block, the formation of monolayers was favored, and, surprisingly, vesicles were formed in water at neutral pH over the entire compositional range. The vesicular structures were extensively characterized by static and dynamic light scattering, small-angle neutron scattering, atomic force microscopy, cryo-transmission electron microscopy, scanning electron microscopy, UV and Fourier transform infrared spectroscopy, and circular dichroism. In contrast to other vesicular structures derived from conventional polymers, the formed polypeptidic vesicles possess the unique feature of being stimuli-responsive to pH and temperature. When the copolypeptides were mixed with plasmid DNA (pDNA), large vesicular structures were also formed. The molecular characterization of the vectors was performed with most of the methods mentioned above, and indicated that the pDNA is both partially condensed on the PLL phase and partially encapsulated inside the vesicle. Consequently, the synthesized vectors combine the advantages of the polylysine-DNA systems to condense large amounts of genes, as well as those of the liposome-DNA systems to better protect the encapsulated DNA. These vectors are expected to present better gene transfection efficiency to the cell nucleus.
authors.affiliation: Chemistry Department, University of Athens, Panepistimiopolis Zografou, Athens, Greece. iatrou@chem.uoa.gr
authors.forename: Hermis
authors.initials: H
authors.lastname: Iatrou
authors.forename: Henrich
authors.initials: H
authors.lastname: Frielinghaus
authors.forename: Sirkku
authors.initials: S
authors.lastname: Hanski
authors.forename: Nikos
authors.initials: N
authors.lastname: Ferderigos
authors.forename: Janne
authors.initials: J
authors.lastname: Ruokolainen
authors.forename: Olli
authors.initials: O
authors.lastname: Ikkala
authors.forename: Dieter
authors.initials: D
authors.lastname: Richter
authors.forename: Jimmy
authors.initials: J
authors.lastname: Mays
authors.forename: Nikos
authors.initials: N
authors.lastname: Hadjichristidis
mesh_terms: D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D008853:Microscopy / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D013057:Spectrum Analysis / Q000379:methods; D013696:Temperature
chemical_list: D010455:Peptides