Synthesis and characterization of poly(amino acid methacrylate)-stabilized diblock copolymer nano-objects

Amino acids constitute one of Nature's most important building blocks. Their remarkably diverse properties (hydrophobic/hydrophilic character, charge density, chirality, reversible cross-linking etc.) dictate the structure and function of proteins. The synthesis of artificial peptides and proteins comprising main chain amino acids is of particular importance for nanomedicine. However, synthetic polymers bearing amino acid side-chains are more readily prepared and may offer desirable properties for various biomedical applications. Herein we describe an efficient route for the synthesis of poly(amino acid methacrylate)stabilized diblock copolymer nano-objects. First, either cysteine or glutathione is reacted with a commercially available methacrylate-acrylate adduct to produce the corresponding amino acid-based methacrylic monomer (CysMA or GSHMA). Well-defined water-soluble macromolecular chain transfer agents (PCysMA or PGSHMA macro-CTAs) are then prepared via RAFT polymerization, which are then chain-extended via aqueous RAFT dispersion polymerization of 2-hydroxypropyl methacrylate. In situ polymerization-induced self-assembly (PISA) occurs to produce sterically-stabilized diblock copolymer nano-objects. Although only spherical nanoparticles could be obtained when PGSHMA was used as the sole macro-CTA, either spheres, worms or vesicles can be prepared using either PCysMA macro-CTA alone or binary mixtures of poly(glycerol monomethacrylate) (PGMA) with either PCysMA or PGSHMA macro-CTAs. The worms formed soft free-standing thermo-responsive gels that undergo degelation on cooling as a result of a worm-to-sphere transition. Aqueous electrophoresis studies indicate that all three copolymer morphologies exhibit cationic character below pH 3.5 and anionic character above pH 3.5. This pH sensitivity corresponds to the known behavior of the poly(amino acid methacrylate) steric stabilizer chains

DNA-copolymers structure formation: beyond self-assembly

This thesis describes various synthetic strategies to generate DNA based amphiphilic copolymers which organized into functional structures. Scholar studies of the mechanism of structure formation were conducted considering the hydrophilic weight fraction as the key factor directing the self-assembly. Various modes of structure formation could be studied, inherent to that parameter i. e., crystallization assisted self-assembly, emulsification and nucleation polymerization. The functionality of the resulting structures as well as their potential application were eventually assessed

Synthesis and Self-Assembly of a DNA Molecular Brush

We report herein on the polymer-crystallization-assisted thiol-ene photosynthesis of an amphiphilic comb/graft DNA copolymer, or molecular brush, composed of a hydrophobic poly(2-oxazoline) backbone and hydrophilic short single-stranded nucleic acid grafts. Coupling efficiencies are above 60% and thus higher as compared with the straight solid-phase-supported synthesis of amphiphilic DNA block copolymers. The DNA molecular brushes self-assemble into sub-micron-sized spherical structures in water as evidenced by light scattering as well as atomic force and electron microscopy imaging. The nucleotide sequences remain functional, as assessed by UV and fluorescence spectroscopy subsequent to isoindol synthesis at the surface of the structures. The determination of a vesicular morphology is supported by encapsulation and subsequent spectroscopy monitoring of the release of a water-soluble dye and spectroscopic quantification of the hybridization efficiency (30% in average) of the functional nucleic acid strands engaged in structure formation: about one-half of the nucleotide sequences are available for hybridization, whereas the other half are hindered within the self-assembled structure. Because speciation between complementary and non complementary sequences in the medium could be ascertained by confocal laser scanning microscopy, the stable self-assembled molecular brushes demonstrate the potential for sensing applications

Polymer-Aptamer Hybrid Emulsion Templating Yields Bioresponsive Nanocapsules

This article describes the synthesis of a DNA–polymer, being the nucleotide sequence an aptamer selected in vitro to target specifically the immunoglobulin E (IgE) protein, an allergy biomarker. Subsequent to coupling to poly(2-alkyl-2-oxazoline) with N-Boc protected amino acid side chains, the resulting amphiphilic DNA–polymer hybrid composed of the water-soluble DNA fragment grafted to the hydrophobic polymer segment can be regarded as a high molecular weight analogue of a surfactant. It is demonstrated that the copolymer–aptamer stabilizes efficiently submicrometer size oil-in-water and water-in-oil emulsions, by dynamic light scattering, microscopy, and reflectometry. Particularly interesting is that the aptamer remains functional after coupling to a polymer backbone, stabilization of the emulsion droplets, and locking of the structure subsequent to cross-linking polymerization. The resulting nanocapsules still target specifically the IgE protein. The biological-stimulus responsiveness of the structures is of high potential for future developments of carriers for sustained and targeted delivery

Formation of DNA-Copolymer Fibrils Through an Amyloid-Like Nucleation Polymerization Mechanism

Conjugation of a hydrophobic poly(2-oxazoline) bearing tertiary amide groups along its backbone with a short single stranded nucleotide sequence results in an amphiphilic comb/graft copolymer, which organizes in fibrils upon direct dissolution in water. Supported by circular dichroism, atomic force microscopy, transmission electron microscopy, and scattering data, fibrils are formed through inter- and intramolecular hydrogen bonding between hydrogen accepting amide groups along the polymer backbone and hydrogen donating nucleic acid grafts leading to the formation of hollow tubes

DNA–Polymer Conjugates: From Synthesis, Through Complex Formation and Self-assembly to Applications

With this review, we aim at achieving a comprehensive and up to date look at the broad topic of DNA conjugates as well as summarizing the important trends in the particular field of materials based on nucleic acids

Effect of the Interaction of the Amyloid β (1–42) Peptide with Short Single-Stranded Synthetic Nucleotide Sequences: Morphological Characterization of the Inhibition of Fibrils Formation and Fibrils Disassembly

The formation of extracellular neuritic plaques in the brain of individuals who suffered from Alzheimer's disease (AD) is a major pathological hallmark. These plaques consist of filamentous aggregates of the amyloid beta (1–42) (Aβ42) proteins. Prevention or reduction of the formation of these fibrils is foreseen as a potential therapeutic approach. In this context, we investigated the interactions between the Aβ42 protein and polyions, in particular short single stranded synthetic nucleotide sequences. The experimental outcomes reported herein provide evidence of the inhibition of amyloid fibril genesis as well as disassembly of existing fibers through electrostatic interaction between the Aβ42 protein and the polyions. Since the polyions and the Aβ42 protein are oppositely charged, the formation of (micellar) inter polyelectrolyte complexes (IPECs) is likely to occur. Since the abnormal deposition of amyloid fibers is an archetype of AD, the outcomes of these investigations, supported by atomic force microscopy imaging in the dry and liquid states, as well as circular dichroism and Fourier transform infrared spectroscopy, are of high interest for the development of future strategies to cure a disease that concerns an ever aging population

Self-assembling DNA–peptide hybrids: morphological consequences of oligonucleotide grafting to a pathogenic amyloid fibrils forming dipeptide

cited By 21International audienceFor the very first time, highly efficient synthesis of DNA-peptide hybrids to scaffold self-assembled nanostructures is described. Oligonucleotide conjugation to the diphenylalanine dipeptide triggers a morphological transition from fibrillar to vesicular structures which may potentially be used as delivery vehicles, since they exhibit pH triggered release. © 2012 The Royal Society of Chemistry