PEGylated polymers for medicine: from conjugation to self-assembled systems.

Synthetic polymers have transformed society in many areas of science and technology, including recent breakthroughs in medicine. Synthetic polymers now offer unique and versatile platforms for drug delivery, as they can be "bio-tailored" for applications as implants, medical devices, and injectable polymer-drug conjugates. However, while several currently used therapeutic proteins and small molecule drugs have benefited from synthetic polymers, the full potential of polymer-based drug delivery platforms has not yet been realized. This review examines both general advantages and specific cases of synthetic polymers in drug delivery, focusing on PEGylation in the context of polymer architecture, self-assembly, and conjugation techniques that show considerable effectiveness and/or potential in therapeutics.

Fluorogenic 1,3-dipolar cycloaddition within the hydrophobic core of a shell cross-linked nanoparticle.

Using either nitroxide mediated polymerization (NMP) or reversible addition fragmentation transfer (RAFT) techniques, novel block copolymers that present terminal acetylenes, in the side chain of the styrenic block, were obtained with narrow polydispersities and targeted molecular weights. For the conversion of these acetylene-functionalized polymers to amphiphilic block copolymers, RAFT techniques were preferred. Mild protection/deprotection chemistries were employed which were compatible with the incorporation of the acetylene functionality in the hydrophobic segment. These acetylene-functionalized, Click-readied amphiphilic block copolymers were then self-assembled and cross-linked to afford shell cross-linked knedel-like (SCK) nanoparticles that contained acetylene groups in the core domain. The hydrodynamic diameters (D(h)) of the block copolymer micelles and nanoparticles were determined by dynamic light scattering (DLS), and the dimensions of the nanoparticles were characterized using tapping-mode atomic force microscopy (AFM) and transmission electron microscopy (TEM). The chemical availability of the Click functionality within the core domain of the SCKs was investigated using the copper(I)-catalyzed 1,3-dipolar fluorogenic cycloaddition with a non-fluorescent 3-azidocoumarin profluorophore to afford intensely fluorescent nanoparticles.

Shell click-crosslinked (SCC) nanoparticles: a new methodology for synthesis and orthogonal functionalization.

A new methodology for the preparation of well-defined core-shell nanoparticles was developed, based upon the employment of a multifunctional crosslinker to coincidently stabilize supramolecular polymer assemblies and imbed into the shell unique chemical functionalities. Amphiphilic diblock copolymers of poly(acrylic acid)(80)-b-poly(styrene)(90) that had been assembled into micelles and partially functionalized throughout the corona with alkynyl groups were utilized as Click-readied nanoscaffolds for the formation of shell Click-crosslinked nanoparticles (SCCs). Divergently grown dendrimers of the zero, first, second, and third generations having increasing numbers of azide terminating groups ((N(3))(2)-[G-0], (N(3))(4)-[G-1], (N(3))(8)-[G-2], and (N(3))(16)-[G-3], respectively) were investigated as crosslinkers via Click reactions with the alkynyl groups to form covalent linkages throughout the block copolymer micelle corona, thus forming a crosslinked shell. The crosslinking reactions were characterized by (1)H NMR and IR spectroscopies, differential scanning calorimetry (DSC), and dynamic light scattering (DLS) measurements. Only the first generation dendrimer ((N(3))(4)-[G-1]) possessed a sufficient balance of polyvalency and water solubility to achieve crosslinking and establish a robust nanostructure. The resulting SCC was further characterized with atomic force microscopy (AFM), transmission electron microscopy (TEM), and analytical ultracentrifugation (AU). The dendritic crosslinker is important as it also allows for the incorporation of excess functionality that can undergo complementary reactions. Within the shell of this SCC the remaining azide termini of the dendrimer crosslinker were then consumed in a secondary Click reaction with an alkynyl-functionalized fluorescein to yield a fluorescently labeled SCC that was characterized with DLS, AFM, TEM, AU, UV-vis, and fluorescent measurements as a function of pH.

An assessment of the effects of shell cross-linked nanoparticle size, core composition, and surface PEGylation on in vivo biodistribution.

Amphiphilic core-shell nanoparticles have drawn considerable interest in biomedical applications. The precise control over their physicochemical parameters and the ability to attach various ligands within specific domains suggest shell cross-linked (SCK) nanoparticles may be used as multi-/polyvalent scaffolds for drug delivery. In this study, the biodistribution of four SCKs, differing in size, core composition, and surface PEGylation, was evaluated. To facilitate in-vivo tracking of the SCKs, the positron-emitting radionuclide copper-64 was used. By using biodistribution and microPET imaging approaches, we found that small diameter (18 nm) SCKs possessing a polystyrene core showed the most favorable biological behavior in terms of prolonged blood retention and low liver accumulation. The data demonstrated that both core composition, which influenced the SCK flexibility and shape adaptability, and hydrodynamic diameter of the nanoparticle play important roles in the respective biodistributions. Surface modification with poly(ethylene glycol) (PEG) had no noticeable effects on SCK behavior.

Antigen-decorated shell cross-linked nanoparticles: synthesis, characterization, and antibody interactions.

Antigen-decorated shell cross-linked knedel-like nanoparticles (SCKs) were synthesized and studied as multivalent nanoscale surfaces from which antibody-binding units were presented in a manner that was designed to approach virus particle surfaces. The SCK nanostructures were fabricated with control over the number of antigenic groups, from mixed micellization of amphiphilic diblock copolymer building blocks that contained either an antigen (2,4-dinitrophenyl) or an ethylpropionate group at the hydrophilic alpha-chain terminus. Amphiphilic diblock copolymers were synthesized by atom transfer radical polymerization of tert-butyl acrylate and methyl acrylate sequentially from either a 2,4-dinitrophenyl-functionalized initiator or ethyl 2-bromopropionate, followed by selective removal of the tert-butyl groups to afford 2,4-dinitrophenyl-poly(acrylic acid)60-b-poly(methyl acrylate)60 (DNP-PAA(60)-b-PMA60) and poly(acrylic acid)70-b-poly(methyl acrylate) (PAA70-b-PMA70). Micelles were assembled via addition of water to THF solutions of the polymers in 0:1, 1:1, and 1:0 molar ratios of DNP-PAA60-b-PMA60 to PAA70-b-PMA70, followed by dialysis against water. The acrylic acid groups of the micelle coronas were partially cross-linked (nominally 50%) with 2,2'-(ethylenedioxy)bis(ethylamine), in the presence of 1-(3'-dimethylaminopropyl)-3-ethylcarbodiimide methiodide. Following extensive dialysis against water, the 0%, 50%, and 100% dinitrophenylated shell cross-linked nanoparticles (DNP-SCKs) were characterized with dynamic light scattering (DLS), transmission electron microscopy (TEM), atomic force microscopy (AFM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), infrared and UV-vis spectroscopies, and analytical ultracentrifugation (AU). The surface accessibility and bioavailability of the DNP units upon the DNP-SCKs were investigated by performing quenching titrations of fluorescein-labeled IgE antibody in solution and degranulation of IgE sensitized RBL-2H3 cells. The DNP antigens proved to be surface-available and able to form multivalent bonds with IgE antibodies, causing degranulation.

Synthesis, characterization, and bioavailability of mannosylated shell cross-linked nanoparticles.

Saccharide-functionalized shell cross-linked (SCK) polymer micelles designed as polyvalent nanoscaffolds for selective interactions with receptors on Gram negative bacteria were constructed from mixed micelles composed of poly(acrylic acid-b-methyl acrylate) and mannosylated poly(acrylic acid-b-methyl acrylate). The mannose unit was conjugated to the hydrophilic chain terminus of the amphiphilic diblock copolymer precursor, from which the SCK nanoparticles were derived, by the growth of the diblock copolymer from a mannoside functionalized atom transfer radical polymerization (ATRP) initiator. Mixed micelle formation between the amphiphilic diblock copolymer and mannosylated amphiphilic diblock copolymer was followed by condensation-based cross-linking between the acrylic acid residues present in the periphery of the polymer micelles to afford SCK nanoparticles. SCKs presenting variable numbers of mannose functionalities were prepared from mixed micelles of controlled stoichiometric ratios of mannosylated and nonmannosylated diblock copolymers. The polymer micelles and SCKs were characterized by dynamic light scattering (DLS), electrophoretic light scattering, atomic force microscopy (AFM), transmission electron microscopy (TEM), and analytical ultracentrifugation (AU). Surface availability and bioactivity of the mannose units were evaluated by interactions of the nanostructures with the model lectin Concanavalin A via DLS studies, with red blood cells (rabbit) via agglutination inhibition assays and with bacterial cells (E. coli) via TEM imaging.