Nucleophile responsive charge-reversing polycations for pDNA transfection.

Polycationic carriers promise low cost and scalable gene therapy treatments, however inefficient intracellular unpacking of the genetic cargo has limited transfection efficiency. Charge-reversing polycations, which transition from cationic to neutral or negative charge, can offer targeted intracellular DNA release. We describe a new class of charge-reversing polycation which undergoes a cationic-to-neutral conversion by a reaction with cellular nucleophiles. The deionization reaction is relatively slow with primary amines, and much faster with thiols. In mammalian cells, the intracellular environment has elevated concentrations of amino acids (∼10×) and the thiol glutathione (∼1000×). We propose this allows for decationization of the polymeric carrier slowly in the extracellular space and then rapidly in the intracellular milleu for DNA release. We demonstrate that in a lipopolyplex formulation this leads to both improved transfection and reduced cytotoxicity when compared to a non-responsive polycationic control.

Chemical signal regulated injectable coacervate hydrogels.

In the quest for stimuli-responsive materials with specific, controllable functions, coacervate hydrogels have become a promising candidate, featuring sensitive responsiveness to environmental signals enabling control over sol-gel transitions. However, conventional coacervation-based materials are regulated by relatively non-specific signals, such as temperature, pH or salt concentration, which limits their possible applications. In this work, we constructed a coacervate hydrogel with a Michael addition-based chemical reaction network (CRN) as a platform, where the state of coacervate materials can be easily tuned by specific chemical signals. We designed a pyridine-based ABA triblock copolymer, whose quaternization can be regulated by an allyl acetate electrophile and an amine nucleophile, leading to gel construction and collapse in the presence of polyanions. Our coacervate gels showed not only highly tunable stiffness and gelation times, but excellent self-healing ability and injectability with different sized needles, and accelerated degradation resulting from chemical signal-induced coacervation disruption. This work is expected to be a first step in the realization of a new class of signal-responsive injectable materials.

Temporally programmed polymer - solvent interactions using a chemical reaction network.

Out of equilibrium operation of chemical reaction networks (CRNs) enables artificial materials to autonomously respond to their environment by activation and deactivation of intermolecular interactions. Generally, their activation can be driven by various chemical conversions, yet their deactivation to non-interacting building blocks remains largely limited to hydrolysis and internal pH change. To achieve control over deactivation, we present a new, modular CRN that enables reversible formation of positive charges on a tertiary amine substrate, which are removed using nucleophilic signals that control the deactivation kinetics. The modular nature of the CRN enables incorporation in diverse polymer materials, leading to a temporally programmed transition from collapsed and hydrophobic to solvated, hydrophilic polymer chains by controlling polymer-solvent interactions. Depending on the layout of the CRN, we can create stimuli-responsive or autonomously responding materials. This concept will not only offer new opportunities in molecular cargo delivery but also pave the way for next-generation interactive materials.

Thioanisole ester based logic gate cascade to control ROS-triggered micellar degradation.

In certain tumor and diseased tissues, reactive oxygen species (ROS), such as H2O2, are produced in higher concentrations than in healthy cells. Drug delivery and release systems that respond selectively to the presence of ROS, while maintaining their stability in "healthy" biological conditions, have great potential as on-site therapeutics. This study presents polymer micelles with 4-(methylthio)phenyl ester functionalities as a ROS-responsive reactivity switch. Oxidation of the thioether moieties triggers ester hydrolysis, exposing a hydrophylic carboxylate and leading to micellar disassembly. At 37 °C, the micelles fall apart on a timescale of days in the presence of 2 mM H2O2 and within hours at higher concentrations of H2O2 (60-600 mM). In the same time frame, the nanocarriers show no hydrolysis in oxidant-free physiological or mildly acidic conditions. This logic gate cascade behavior represents a step forward to realize drug delivery materials capable of selective response to a biomarker input.

Fuel-driven macromolecular coacervation in complex coacervate core micelles.

Fuel-driven macromolecular coacervation is an entry into the transient formation of highly charged, responsive material phases. In this work, we used a chemical reaction network (CRN) to drive the coacervation of macromolecular species readily produced using radical polymerisation methods. The CRN enables transient quaternization of tertiary amine substrates, driven by the conversion of electron deficient allyl acetates and thiol or amine nucleophiles. By incorporating tertiary amine functionality into block copolymers, we demonstrate chemical triggered complex coacervate core micelle (C3M) assembly and disassembly. In contrast to most dynamic coacervate systems, this CRN operates at constant physiological pH without the need for complex biomolecules. By varying the allyl acetate fuel, deactivating nucleophile and reagent ratios, we achieved both sequential signal-induced C3M (dis)assembly, as well as transient non-equilibrium (dis)assembly. We expect that timed and signal-responsive control over coacervate phase formation at physiological pH will find application in nucleic acid delivery, nano reactors and protocell research.

Linear Coordination Polymer Synthesis from Bis-Catechol Functionalized RAFT Polymers.

Catechol-Fe(III) complexes contain some of the strongest known metal-chelate coordination bonds. Despite this, they have until now not been utilized in (polymeric linker) linear coordination polymer (LCP) synthesis. With the view of generating catechol end-functional polymers, a new, symmetrical bis-catechol functionalized trithiocarbonate reversible addition fragmentation chain transfer (RAFT) agent is synthesized (CatDMAT). Acrylamide (AM) and dimethylacrylamide (DMA) polymerizations are conducted with CatDMAT using direct photoactivation RAFT polymerization to yield bis-catechol end-functionalized homo- and block-copolymers of molecular weight 10-15 kDa. Catechol-Fe(III) LCPs are successfully formed from the telechelic catechol polymers by bis-complexation to Fe(III). The tetrahedral bis-complex is detected by UV-vis spectroscopy (λmax  = 570 nm), while increases in relative viscosity and Mn,GPC over their respective uncomplexed polymers confirm the occurrence of supramolecular polymerization. The catechol-LCPs are shown to undergo oxidation and crosslinking in aqueous solution after 24 h.

Ultra-high molecular weight linear coordination polymers with terpyridine ligands.

Ultra-high molecular weight (UHMW, M n > 1000 kDa) polymeric drift control adjuvants (DCAs) for agricultural spraying are prone to mechanical degradation and rapidly lose performance. To overcome this, we have designed linear coordination polymers (LCPs) composed of 400 kDa telechelic bis-terpyridine end-functionalised polyacrylamide units, which 'self-heal' upon shearing through reformation of coordination bonds. After addition of Fe(ii) to dilute aqueous solutions of the terpyridine telechelics, UHMW LCPs were obtained as demonstrated by UV-vis spectroscopy, MALS GPC and intrinsic viscosity measurements. Importantly, these UHMW LCPs were shown to function as effective DCAs, reducing the formation of fine 'driftable' droplets during spray testing at concentrations as low as 100 ppm. Following mechanically-induced coordination bond-scission, the UHMW LCPs were found to recover up to 90% of their performance compared to un-sheared samples, at a rate dependent on the transition metal ion used to form the complex.

Light-Induced RAFT Single Unit Monomer Insertion in Aqueous Solution-Toward Sequence-Controlled Polymers.

First report on the sequential, visible light-initiated, single unit monomer insertion (SUMI) of N,N-dimethylacrylamide (DMAm) into the reversible addition fragmentation chain transfer (RAFT) agent, 4-((((2-carboxyethyl)thio)carbonothioyl)thio)-4-cyanopentanoic acid (CTA1 ), in aqueous solution is provided. The specificity for SUMI over formation of higher oligomers and/or RAFT agent-derived by-products is higher for longer irradiation wavelengths. Red light provides the cleanest product (selective SUMI), showing a linear pseudo-first order kinetic profile to high (>80%) conversion, but also the slowest reaction rate. Blue light provides a relatively rapid reaction, but also gives some by-products (<2%) and the kinetic profile displays a conversion plateau at >65% conversion. Higher specificity with red light is attributed to CTA1 absorbing at longer wavelengths than the SUMI product, which allows selective excitation of CTA1 . The use of a higher reaction temperature (65 °C vs ambient) results in a higher reaction rate and a reduction in oligomer formation.