Diblock Micelles Research Articles

Blended Block Polycation Micelles Enhance Antisense Oligonucleotide Delivery.

Nucleic acid-based medicines and vaccines are becoming an important part of our therapeutic toolbox. One key genetic medicine is antisense oligonucleotides (ASOs), which are short single-stranded nucleic acids that downregulate protein production by binding to mRNA. However, ASOs cannot enter the cell without a delivery vehicle. Diblock polymers containing cationic and hydrophobic blocks self-assemble into micelles that have shown improved delivery compared to linear nonmicelle variants. Yet synthetic and characterization bottlenecks have hindered rapid screening and optimization. In this study, we aim to develop a method to increase throughput and discovery of new micelle systems by mixing diblock polymers together to rapidly form new micelle formulations. We synthesized diblocks containing an n-butyl acrylate block chain extended with cationic moieties amino ethyl acrylamide (A), dimethyl amino ethyl acrylamide (D), or morpholino ethyl acrylamide (M). These diblocks were then self-assembled into homomicelles (A100, D100, and M100)), mixed micelles comprising 2 homomicelles (MixR%+R'%), and blended diblock micelles comprising 2 diblocks blended into one micelle (BldR%R'%) and tested for ASO delivery. Interestingly, we observed that mixing or blending M with A (BldA50M50 and MixA50+M50) did not improve transfection efficiency compared to A100; however, when M was mixed with D, there was a significant increase in transfection efficacy for the mixed micelle MixD50+M50 compared to D100. We further examined mixed and blended D systems at different ratios. We observed a large increase in transfection and minimal change in toxicity when M was mixed with D at a low percentage of D incorporation in mixed diblock micelles (i.e., BldD20M80) compared to D100 and MixD20+M80. To understand the cellular mechanisms that may result in these differences, we added proton pump inhibitor Bafilomycin-A1 (Baf-A1) to the transfection experiments. Formulations that contain D decreased in performance in the presence of Baf-A1, indicating that micelles with D rely on the proton sponge effect for endosomal escape more than micelles with A. This result supports our conclusion that M is able to modulate transfection of D, but not with A. This research shows that polymer blending in a manner similar to that of lipids can significantly boost transfection efficiency and is a facile way to increase throughput of testing, optimization, and successful formulation identification for polymeric nucleic acid delivery systems.

Core-crosslinked micelles with a poly-anionic poly(styrene sulfonate)-based outer shell made by RAFT polymerization

Core-crosslinked micelles (CCMs) with a permanently charged anionic shell of sodium poly(styrene sulfonate) (PSS–Na+) chains have been developed by a convergent three-step one-pot RAFT polymerization starting with 4-cyano-4-thiothiopropylsulfanyl pentanoic acid (CTPPA, R0-SC(S)SnPr) as chain transfer agent, which includes polymerization-induced self-assembly (PISA) in the second step. The final crosslinking step of the R0-(SS-Na+)x-b-Sty-SC(S)SnPr diblock chains was carried out with diethylene glycol dimethacrylate (DEGDMA), either diluted into styrene (DEGDMA/St = 10:90) or neat. The morphology of the intermediate micelles and of the final CCM particles was studied by dynamic light scattering (DLS) and transmission electron microscopy (TEM) as a function of the relative degrees of polymerizations for the shell, core, and crosslinking blocks. Conformational instability was observed for the aqueous dispersions of the diblock micelles, especially with the shorter PSt blocks, yielding equilibria with single chains and large agglomerates. However, crosslinking led to well-defined CCMs with narrow size distributions for the optimized polymer compositions. The DLS analysis in a compatibilizing 60:40 THF/H2O solvent mixture demonstrates that the final step of the polymer synthesis has quantitatively crosslinked all diblock arms.

Stimuli‐responsive amphiphilic PDMAEMA‐b‐PLMA copolymers and their cationic and zwitterionic analogs

ABSTRACTIn this work, the synthesis and characterization of novel amphiphilic diblock copolymers of poly(2‐dimethylamino ethyl methacrylate)‐b‐poly(lauryl methacrylate), PDMAEMA‐b‐PLMA, using the reversible addition‐fragmentation chain transfer (RAFT) polymerization technique, are reported. The diblocks were successfully derivatized to cationic and zwitterionic block polyelectrolytes by quaternization and sulfobetainization of the PDMAEMA block, respectively. Furthermore, their molecular and physicochemical characterization was performed by using characterization techniques such as NMR and FTIR, size exclusion chromatography, light scattering techniques, and transmission electron microscopy. The structure of the diblock micelles, their behavior, and properties in aqueous solution were investigated under the effect of pH, temperature, and ionic strength, as PDMAEMA and its derivatives are stimuli‐responsive polymers and exhibit responses to variations of at least one of these physicochemical parameters. These new families of stimuli‐responsive block copolymers respond to changes of their environment giving interesting nanostructures, behavioral motifs, and properties, rendering them useful as nanocarriers for drug delivery and gene therapy. © 2017 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2018, 56, 598–610

Poly(lactic acid)/poly(ethylene glycol) block copolymer based shell or core cross-linked micelles for controlled release of hydrophobic drug

The syntheses, structures, and drug release behavior of shell or core cross-linked poly(lactic acid)/poly(ethylene glycol) block copolymer micelles.

Synthesis of dual thermoresponsive ABA triblock copolymers by both living cationic vinyl polymerization and RAFT polymerization using a dicarboxylic RAFT agent

To elucidate a difference of micelles formed from AB di- and ABA tri-block copolymers, PMOVE–PNIPAM–PMOVE triblock copolymers were synthesized by both living cationic polymerization of MOVE and RAFT polymerization of NIPAM using a dicarboxylic RAFT agent, where MOVE is 2-methoxyethyl vinyl ether, NIPAM is N-isopropylacrylamide. The ABA triblock copolymers consist of cationically polymerizable monomer (A), MOVE, and radically polymerizable monomer (B), NIPAM. The key to success in the triblock copolymerization is to utilize a dicarboxylic RAFT agent/SnBr4 initiation system for living cationic polymerization. The cationic polymerization was simultaneously initiated from two protons per one dicarboxylic RAFT agent. After the cationic polymerization, the polymerization was quenched and then the RAFT carboxylate counter anion was concurrently recovered at ca. 86% efficiency, followed by the RAFT polymerization. The resulting PMOVE–PNIPAM–PMOVE triblock copolymers contain two thermoresponsive blocks. The resulting block copolymers reversibly formed or deformed micellar assemblies in water. When the chain length of the core-forming block, PNIPAM, is increased, larger micelles were invariably obtained. The triblock micellar size was larger than that of the corresponding diblock micelle. The micelles at the concentrated solution formed physical gel, and the minimum gelation concentration of the triblock micelles was lower than that of the corresponding diblock micelles.

Pressure Jump Kinetics of Disorder to BCC Ordering in Diblock Copolymer Micelles in a Selective Solvent

We present time-resolved small-angle X-ray scattering (SAXS) measurements of the kinetics of a barotropic disorder–order transition in polystyrene–polyisoprene (SI) diblock micelles in diethyl phthalate (DEP), a styrene selective solvent. Because of the fast equilibration in a pressure jump we were able to observe the initial induction stage during which the short-range order of the micellar fluid increases, followed by a sharp first-order nucleation and growth of the BCC phase and eventual late stage coarsening. The time evolution of intensity from individual diffraction spots of a crystallite was obtained from the analysis of 2-dimensional SAXS patterns and shows late stage power-law growth. The time evolution of the SAXS intensity profiles was analyzed by fitting the contribution to the scattering from the ordered Bragg peaks by Gaussians and analyzing the contribution to the scattering from the disordered micellar liquid in terms of the Percus–Yevick interacting hard-sphere model. The time evolution of the micellar structural parameters obtained from this analysis reveals that the disorder to order transition occurs when the volume fraction of micelles reaches 0.53, and the effective hard sphere radius and volume fraction of micelles increases upon ordering while the core radius is unaffected. In both the ordered and disordered states the hard sphere radius and volume fraction of micelles increase with increasing pressure indicating increased swelling of the micellar corona. In the ordered state there is considerable penetration of the corona chains from neighboring micelles.

Aqueous reversible addition‐fragmentation chain transfer dispersion polymerization of thermoresponsive diblock copolymer assemblies: Temperature directed morphology transformations

AbstractIn this work, we demonstrate the formation of various 3D structures formed by a structural reorganization; a process not governed by self‐assembly. First, we packaged well‐defined diblocks, thermoresponsive poly(N‐isopropylacrylamide‐b‐styrene) or P(NIPAM‐b‐STY), into spherical particles made in situ using a reversible addition‐fragmentation chain transfer (RAFT) nanoreactor technique in water to obtain high polymer solids. The resultant spheres reorganized through a temperature stimulus to form equilibrium and kinetically trapped structures; we denote this process as temperature directed morphology transformation (TDMT). Cylinder and vesicle structures, other more unusual loop, buckled sphere and cauliflower (via ultrasound) structures were observed below the lower critical solution temperature (LCST) of PNIPAM. These structures were produced efficiently, rapidly, reproducibly at high polymer solids and can be stored for years in solution or be freeze‐dried and rehydrated without a change in structure, which becomes important in biological applications. We generated the first phase diagram for the TDMT changes of thermoresponsive diblock micelles in concentrated solutions (> 7% polymer solids) produced from narrow molecular weight diblock copolymers (PDI ∼ 1.1). The sphere/cylinder and cylinder/vesicle transition in the phase diagram were surprisingly found to be predominantly dictated by the length of the hydrophilic block, poly(N‐isopropylacrylamide). © 2012 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2012

“Design of Poly(ethylene glycol)-Polycaprolactone Diblock Micelles with RGD Targeting Ligands and Embedded Iron Oxide Nanoparticles for Thermally-activated Release”

ABSTRACTA thermally-activated micelle consisting of a crystallizable poly(caprolactone), PCL, core and a poly(ethylene glycol), PEG, corona was developed to contain magnetic nanoparticles and anti-cancer agent doxorubicin as well as display a targeting RGD peptide. This system has the potential to target cancer cells, deliver combination hyperthermia and chemotherapy, and offer magnetic resonance imaging contrast. The micelles self-assemble in aqueous solutions and form a crystalline core with a melting transition ranging from 40 to 50 °C, depending on the length of the PCL blocks, with dynamic light scattering showing micelle sizes typically ranging from 20 to 100 nm, depending on block lengths and added drug or nanoparticles. The micelles become unstable as they are heated above their melting point, creating a thermally-activated drug release mechanism. By adding magnetite (Fe3O4) nanoparticles into the PCL core, the micelles can be heated using an externally applied AC magnetic field to induce hyperthermia in combination with the thermally-activated drug release. The polymers and magnetic nanoparticles (MNPs) were synthesized and characterized in our laboratories. The melting transitions of the PCL micelle cores were investigated using microcalorimetry. The heating of nanoparticles and magnetomicelles was conducted using a custom-built hyperthermia coil capable of generating fields of several hundred Oersteds at frequencies ranging from 50 to 450 kHz. Heating of MNPs was maximized at high field intensities. RGD peptides were attached to the PEG corona using maleimide chemistry, and the ability of the RGD-derivatized micelles to target integrin-expressing cells was investigated using fluorescent dye PKH26 to identify the localization of micelles in cultured human kidney (293) cells in vitro. The crystallizable (and meltable) cores in these micelles were designed to overcome drug leakage common in liposome systems and release the drug on demand after a period of time for localization to integrin receptors.

Stimulated release of photosensitizers from graft and diblock micelles for photodynamic therapy

To understand the effect of photosensitizer (PS) release from graft copolymer based micelles in photodynamic therapy (PDT), the two pH-sensitive and non-pH-sensitive graft copolymers, (poly(N-vinyly caprolactam)-g-poly( d, l-lactide) and poly(N-vinyly caprolactam-co-N-vinyl imidazole)-g-poly( d, l-lactide)), were synthesized and utilized for the encapsulation of protoporphyrin IX (PPIX) for in vitro and in vivo PDT studies. Photochemical internalization (PCI) was utilized to study the localization of pH- and non-pH-sensitive micelles uptake in the lysosome. After non-toxic light treatment, PPIX was found in the nucleus with pH-sensitive micelles, while PPIX was still localized in the lysosomal organism with the non-pH-sensitive micelles, as observed by confocal microscopy. Because the formation of singlet oxygen was observed for the block and graft micelles, dramatic differences in the cell viability could be ascribed to the damage occurring at the region where the PPIX was located. An in vivo study revealed that PPIX-loaded graft and diblock micelles presented prolonged blood circulation and enhanced tumor targeting ability. The PPIX released from g-CIM micelles on tumor site was further proved by ex vivo confocal image. In addition, non-pH-sensitive micelle-treated mice showed a better repression of tumor growth than PPIX-treated mice, which was likely due to the larger amount of PS localized in the tumor region still exhibiting therapeutic effects. Finally, effective PDT-induced inhibition of tumor growth was found in pH-sensitive micelle-treated mice. This work provides insight into PS-loaded graft and diblock micelles for the PDT of tumors.

A Cremophor-Free Formulation for Tanespimycin (17-AAG) Using PEO-b-PDLLA Micelles: Characterization and Pharmacokinetics in Rats

Tanespimycin (17-allylamino-17-demethoxygeldanamycin or 17-AAG) is a promising heat shock protein 90 inhibitor currently undergoing clinical trials for the treatment of cancer. Despite its selective mechanism of action on cancer cells, 17-AAG faces challenging issues due to its poor aqueous solubility, requiring formulation with Cremophor EL (CrEL) or ethanol (EtOH). Therefore, a CrEL-free formulation of 17-AAG was prepared using amphiphilic diblock micelles of poly(ethylene oxide)-b-poly(D,L-lactide) (PEO-b-PDLLA). Dynamic light scattering revealed PEO-b-PDLLA (12:6 kDa) micelles with average sizes of 257 nm and critical micelle concentrations of 350 nM, solubilizing up to 1.5 mg/mL of 17-AAG. The area under the curve (AUC) of PEO-b-PDLLA micelles was 1.3-fold that of the standard formulation. The renal clearance (CL(renal)) increased and the hepatic clearance (CL(hepatic)) decreased with the micelle formulation, as compared to the standard vehicle. The micellar formulation showed a 1.3-fold increase in the half-life (t(1/2)) of the drug in serum and 1.2-fold increase in t(1/2) of urine. As expected, because it circulated longer in the blood, we also observed a 1.7-fold increase in the volume of distribution (V(d)) with this micelle formulation compared to the standard formulation. Overall, the new formulation of 17-AAG in PEO-b-PDLLA (12:6 kDa) micelles resulted in a favorable 150-fold increase in solubility over 17-AAG alone, while retaining similar properties to the standard formulation. Our data indicates that the nanocarrier system can retain the pharmacokinetic disposition of 17-AAG without the need for toxic agents such as CrEL and EtOH.

Unraveling the equilibrium chain exchange kinetics of polymeric micelles using small-angle neutron scattering – architectural and topological effects

In this paper, we present a study of micellar structures formed by poly(styrene)-poly(butadiene) (PS10-PB10; the numbers indicate the molecular weight in kg mol−1) diblock copolymers and PB10-PS20-PB10 triblock copolymers in different n-alkane solvents. Particular emphasis is placed on the dynamic properties of these micelles under equilibrium which are studied using a novel time-resolved small-angle neutron scattering technique. The results show that the structures of the micelles are very similar for both the diblock and triblock copolymers, which allows a direct comparison of the dynamic properties. A novel logarithmic relaxation is found for both the triblock and the diblock micelles which is not consistent with theoretical expectations. However, for the diblock micelles, the relaxation kinetics seem to approach the rate and the single exponential decay predicted by Halperin & Alexander [Macromolecules, (1989), 22, 2403–2412] when the micellar cores are strongly swollen with solvent. For the triblock micelles a logarithmic relaxation is found for all cases as an effect of additional topological knots present even in highly swollen micellar cores. This behavior is assigned to an increased coupling of chain motion within the dense confined core – an effect which seems to vanish in diblock micelles when the core is sufficiently swollen.

Characterization and comparison of diblock and triblock amphiphilic copolymers of poly(δ-valerolactone)

Three types of pegylated amphiphilic copolymers of poly(δ-valerolactone) (PVL) were copolymerized with methoxy poly(ethylene glycol) (MePEG) and poly(ethylene glycol) (PEG4000 and PEG10,000), respectively. Pegylation of PVL allowed copolymers possessing amphiphilic property and efficiently self-assembled to form micelles with a low critical micelle concentration (CMC) in the range of 10−7–10−8M. The average molecular weight of copolymers was in the range of 10,000–20,000 Da, and the polydispersity of copolymers was about 1.7–1.8. Higher mobility of low molecular weight PEG (i.e., MePEG and PEG4000) than high molecular weight PEG10,000 allowed valerolactone ring opening more efficient in terms of PVL/MePEG and PVL/PEG4000 copolymers possessing longer chain length in hydrophobic domain. Pegylated PVL with low CMC and triblock structure was preferred to encapsulate drug during micelle formation. Although all of these amphiphilic copolymers exhibited controlled release character, the micelles formed by triblock copolymer possessed a more stable core-shell conformation than that by diblock copolymer, and resulted in the release of drug from triblock micelles slower than that from diblock micelles. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 1836–1841, 2006

Reversible Morphological Transitions of Polystyrene-b-polyisoprene Micelles

Reversible morphological transitions of diblock copolymer micelles in dilute solutions were monitored by light scattering and atomic force microscopy for two different polymer samples near the calculated morphological boundaries. These transitions were induced solely by temperature changes. At 25 °C, a sample of polystyrene-b-polyisoprene diblock micelles with a polystyrene block of 20.6 kDa and a polyisoprene block of 6 kDa was observed to form cylindrical micelles in heptane, a selective solvent for polyisoprene. Upon heating to 35 °C, the sample adopted a spherical micelle morphology. When the sample was cooled back to 25 °C, cylindrical micelles were once again observed. In addition, a reversible transition from vesicles to cylindrical micelles, upon heating from 25 to 40 °C, was observed for a second diblock sample with the same polystyrene block (20.6 kDa) and a shorter polyisoprene block of 4.3 kDa. The change in morphology upon heating was found to be much faster then the reverse process upon cooling.

Dynamics of core-shell particles in concentrated suspensions.

The dynamics of soft colloidal particles in a concentrated suspension was studied by two color dynamic light scattering. The short-time diffusion coefficient, D(s)(q), revealed a weak slowing down around the peak of S(q) in contrast to the much stronger dependence observed in hard sphere systems, and the absence of any slowing down in ultrasoft diblock micelles. The low-q limit of the diffusion coefficient, D(s)(q=0), reflecting the cooperative nature of the dynamics was found to decrease from its dilute limit value in contrast to the weak increase observed in hard sphere systems. The behavior of D(s)(q) reflects distinct polymer-layer-induced hydrodynamic effects present in concentrated suspensions of core-shell particles.

Supramolecular Photophysics of Self-Assembled Block Copolymers Containing Luminescent Conjugated Polymers

Luminescent micellar aggregates and spherical vesicles self-assembled from polyquinoline−polystyrene diblock and polyquinoline−polystyrene−polyquinoline triblock copolymers were investigated by photoluminescence excitation and emission spectroscopies and fluorescence microscopy. In both the diblock micelles and triblock vesicles, new absorption and emission features characteristic of the supramolecular morphology in solution and the solid state were observed. J-like aggregation of the conjugated polymer blocks within the self-organized assemblies explain the observed supramolecular photophysics. The large red shift between aggregate luminescence in the solid state compared to solution is attributed to excimer emission. Self-assembling block copolymers containing conjugated polymer blocks thus constitute a promising route to functional supramolecular materials with tailorable optoelectronic and photonic properties.

Umbrella-Star Block Copolymers

Abstract The large scale architecture of polymers is recognized to create special properties in functional polymers. One of the more striking examples is the ability of certain diblock copolymers to associate into micelles in a selective solvent for one of the blocks.(1) The insoluble block forms a dense core that is kept in solution by a soluble corona. The diblock micelle has the overall conformation of a star polymer.(2,3) However, the self-association of block copolymers into micelles depends strongly on the chemical nature of the two blocks, on the respective sizes of the blocks and on the selective solvent. In general, it is observed that the molecular weight of the insoluble block must be kept low in order to form micelles. This is especially true when water or a polar solvent is used as the selective good solvent.(1,4) Otherwise, a cloudy unstable two phase system is produced. The association number of diblocks in micelles varies considerably and makes their systematic study difficult. The association number is known to depend on the preparation method and on the presence of traces of a cosolvent.(4) Star block copolymers whose structure is fixed by covalent bonds are obvious models for diblock micelles.(5,6) In this paper we describe the synthesis and some properties of umbrella-star block copolymers. This architecture allows us to influence the chain density in the corona and to study its effect on the solution properties of the polymers. These polymers are direct models for micelles prepared from umbrella polymers(7) or mixed arm star polymers.(8)