Postpolymerization Modification of Poly(2-vinyl-4,4-dimethyl azlactone) as a Versatile Strategy for Drug Conjugation and Stimuli-Responsive Release.

Postpolymerization modification of highly defined "scaffold" polymers is a promising approach for overcoming the existing limitations of controlled radical polymerization such as batch-to-batch inconsistencies, accessibility to different monomers, and compatibility with harsh synthesis conditions. Using multiple physicochemical characterization techniques, we demonstrate that poly(2-vinyl-4,4-dimethyl azlactone) (PVDMA) scaffolds can be efficiently modified with a coumarin derivative, doxorubicin, and camptothecin small molecule drugs. Subsequently, we show that coumarin-modified PVDMA has a high cellular biocompatibility and that coumarin derivatives are liberated from the polymer in the intracellular environment for cytosolic accumulation. In addition, we report the pharmacokinetics, biodistribution, and antitumor efficacy of a PVDMA-based polymer for the first time, demonstrating unique accumulation patterns based on the administration route (i.e., intravenous vs oral), efficient tumor uptake, and tumor growth inhibition in 4T1 orthotopic triple negative breast cancer (TNBC) xenografts. This work establishes the utility of PVDMA as a versatile chemical platform for producing polymer-drug conjugates with a tunable, stimuli-responsive delivery.

Multi-Stimuli Responsive Sequence Defined Multi-Arm Star Diblock Copolymers for Controlled Drug Release.

Star-shaped polymeric materials provide very high efficiency toward various engineering and biomedical applications. Due to the absence of straightforward and versatile synthetic protocols, the synthesis of sequence-defined star-shaped (co)polymers has remained a major challenge. Here, a facile approach is developed that allows synthesis of a series of unprecedented discrete, multifunctional four-, six-, and eight-arm star-shaped complex macromolecular architectures based on a well-defined triple (thermo/pH/light)-stimuli-responsive poly(N-isopropylacrylamide)-block-poly(methacrylic acid)-umbelliferone (PNIPAM-b-PMAA)n-UMB diblock copolymer, based on temperature responsive PNIPAM segment, pH-responsive PMAA segment, and photoresponsive UMB end groups. Thus, developed star-shaped copolymers self-assemble in water to form spherical nanoaggregates of diameter 90 ± 20 nm, as measured by FESEM. The star-shaped copolymer's response to external stimuli has been assessed against changes in temperature, pH, and light irradiation. The star-shaped copolymer was employed as a nanocarrier for pH responsive release of an anticancer drug, doxorubicin. This study opens up new avenues for efficient star-shaped macromolecular architecture construction for engineering and biomedical applications.

Dual-Responsive Glycopolymers for Intracellular Codelivery of Antigen and Lipophilic Adjuvants.

Traditional approaches to vaccines use whole organisms to trigger an immune response, but they do not typically generate robust cellular-mediated immunity and have various safety risks. Subunit vaccines composed of proteins and/or peptides represent an attractive and safe alternative to whole organism vaccines, but they are poorly immunogenic. Though there are biological reasons for the poor immunogenicity of proteins and peptides, one other key to their relative lack of immunogenicity could be attributed to the poor pharmacokinetic properties of exogenously delivered proteins and peptides. For instance, peptides often aggregate at the site of injection and are not stable in biological fluids, proteins and peptides are rapidly cleared from circulation, and both have poor cellular internalization and endosomal escape. Herein, we developed a delivery system to address the lack of protein immunogenicity by overcoming delivery barriers as well as codelivering immune-stimulating adjuvants. The glycopolymeric nanoparticles (glycoNPs) are composed of a dual-stimuli-responsive block glycopolymer, poly[2-(diisopropylamino)ethyl methacrylate]-b-poly[(pyridyl disulfide ethyl methacrylate)-co-(methacrylamidoglucopyranose)] (p[DPA-b-(PDSMA-co-MAG)]). This polymer facilitates protein conjugation and cytosolic release, the pH-responsive release of lipophilic adjuvants, and pH-dependent membrane disruption to ensure cytosolic delivery of antigens. We synthesized p[DPA-b-(PDSMA-co-MAG)] by reversible addition-fragmentation chain transfer (RAFT) polymerization, followed by the formation and physicochemical characterization of glycoNPs using the p[DPA-b-(PDSMA-co-MAG)] building blocks. These glycoNPs conjugated the model antigen ovalbumin (OVA) and released OVA in response to elevated glutathione levels. Moreover, the glycoNPs displayed pH-dependent drug release of the model hydrophobic drug Nile Red while also exhibiting pH-responsive endosomolytic behavior as indicated by a red blood cell hemolysis assay. GlycoNPs coloaded with OVA and the toll-like receptor 7/8 (TLR-7/8) agonist Resiquimod (R848) activated DC 2.4 dendritic cells (DCs) significantly more than free OVA and R848 and led to robust antigen presentation of the OVA epitope SIINFEKL on major histocompatibility complex I (MHC-I). In sum, the dual-stimuli-responsive glycopolymer introduced here overcomes major protein and peptide delivery barriers and could vastly improve the immunogenicity of protein-based vaccines.

Comparative Investigation of the Hydrolysis of Charge-Shifting Polymers Derived from an Azlactone-Based Polymer.

Poly 2-vinyl-4,4-dimethylazlactone (PVDMA) has received much attention as a "reactive platform" to prepare charge-shifting polycations via post-polymerization modification with tertiary amines that possess primary amine or hydroxyl reactive handles. Upon hydrolysis of the resulting amide or ester linkages, the polymers can undergo a gradual transition in net charge from cationic to anionic. Herein, a systematic investigation of the hydrolysis rate of PVDMA-derived charge-shifting polymers is described. PVDMA is modified with tertiary amines bearing either primary amine, hydroxyl, or thiol reactive handles. The resulting polymers possess tertiary amine side chains connected to the backbone via amide, ester, or thioester linkages. The hydrolysis rates of each PVDMA derivative are monitored at 25 and 50 °C at pH values of 5.5, 7.5, and 8.5, respectively. While the hydrolysis rate of the amide-functionalized PVDMA is negligible over the period investigated, the hydrolysis rates of the ester- and thioester-functionalized PVDMA increase with increasing temperature and pH. Interestingly, the hydrolysis rate of the thioester-functionalized PVDMA appears to be more rapid than the ester-functionalized PVDMA at all pH values and temperatures investigated. It is believed that these results can be utilized to inform the future preparation of PVDMA-based charge-shifting polymers for biomedical applications.

l-Histidine-Derived Smart Antifouling Biohybrid with Multistimuli Responsivity.

A novel dual pH/thermoresponsive amphiphilic poly(histidine methacrylamide)-block-hydroxyl-terminated polybutadiene-block-poly(histidine methacrylamide) (PHisMAM-b-PB-b-PHisMAM) triblock copolymer biohybrid, composed of hydrophobic PB and ampholytic PHisMAM segments, is developed via direct switching from living anionic polymerization to recyclable nanoparticle catalyst-mediated reversible-deactivation radical polymerization (RDRP). The transformation involved in situ postpolymerization modification of living polybutadiene-based carbanionic species, end-capped with ethylene oxide, into dihydroxyl-terminated polybutadiene and a subsequent reaction with 2-bromo-2-methylpropionyl bromide resulting in a telechelic ATRP macroinitiator (Br-PB-Br). Br-PB-Br was used to mediate RDRP of an l-histidine-derived monomer, HisMAM, yielding a series of PHisMAM-b-PB-b-PHisMAM triblock copolymers. The copolymer's stimuli response was assessed against pH and temperature changes. The copolymer is capable of switching among its zwitterionic, anionic, and cationic forms and exhibited unique antifouling properties in its zwitterionic form. These novel triblock copolymers are expected to be show promising potential in biomedical applications.

Anisotropic and Self-Healing Copolymer with Multiresponsive Capability via Recyclable Alloy-Mediated RDRP.

A novel triple stimuli sensitive block copolymer is prepared by magnetically separable and reusable (up to multiple cycles) Ni-Co alloy nanoparticles mediated reversible deactivation radical polymerization (RDRP) at 25 °C, that responds to changes in temperature, pH, and light. Design of this block copolymer constitutes a temperature-sensitive N-isopropylacrylamide (NIPAM), an acid-sensitive lysine methacrylamide (LysMAM), and a light responsive umbelliferone (UMB) end group. The stimuli response, in response to one stimulus as well as combinations of stimuli, has been evaluated. Responsiveness to light allows the construction of self-healing materials. Density functional theory calculations rationalize the underlying mechanism of the polymerization.