The case for poly(ylides) as a class of charge-neutral, hydrophilic polymers with applications in biomaterials science.

Many applications of biomaterials require hydrophilic polymers as building blocks, including hydrogels and nanomedicinal devices. Besides enabling sufficient swelling properties in aqueous environments, hydrophilic polymers provide hydration layers, which are considered a major requirement when designing non-fouling surfaces and materials. For the last few decades, polyethylene glycol has been seen as the gold standard for such applications. However, reports on its stability and immunogenicity have urged chemists to identify alternatives with comparable or superior properties. In addition to biopolymers, zwitterionic polymers have gained increasing attention by effectively offering an overall charge-neutral scaffold capable of forming strong hydration layers. Driven by an enhanced understanding of the structure-property relationship of zwitterionic materials, poly(ylides) have emerged as a new class of hydrophilic and charge-neutral polymers. By having the negative charge adjacent to the positive charge, ylides offer not only a minimal dipole moment but also maintain their overall charge-neutral nature. Despite some early reports on their synthesis during the 1980s, polymeric ylides were largely overlooked as a class of polymers, and their utility as unique hydrophilic building blocks for the design of biomaterials and nanomedicinal tools remained elusive. In recent years, several groups have reported N-oxide and carbon-centered ylide-based polymers as highly effective building blocks for the design of antifouling materials and nanomedicines. Here, by reviewing recent progress and understanding of structure-property relationships, arguments are provided explaining why polymeric ylides should be classified as a standalone class of hydrophilic polymers. Consequently, the author concludes that the term 'poly(ylide)' or 'polymeric ylides' should be routinely used to adequately describe this emerging class of polymers.

Rapid, traceless and facile peptide cyclization enabled by tetrazine-thiol exchange.

Cyclic peptides offer many advantages compared to their linear counterparts, including prolonged stability within the biological environment and enhanced binding affinity. Typically, peptides are cyclized by forming an amide bond, either on-resin or in solution, through extensive use of orthogonal protecting groups or chemoselective ligation strategies, respectively. Here, we show that the chemoselective tetrazine-thiol exchange is a powerful tool for rapid in situ cyclization of peptides without the need for additional activation reagents or extensive protecting group reshuffling. The reaction between N-terminal sulfide-bearing unsymmetric tetrazines and internal cysteines occurs spontaneously within a mildly acidic environment (pH 6.5) and is of traceless nature. The rapidly available unsymmetric sulfide tetrazine building blocks can be incorporated on resin using standard solid-phase peptide synthesis protocols and are orthogonal to trifluoroacetic acid cleavage conditions. The cyclized peptides display high stability, even when incubated with a large excess of free thiols. Due to its traceless and mild nature, we expect that the tetrazine-thiol exchange will be of high value for the in situ formation of cyclic peptide libraries, thus being applicable in drug discovery and development.

Revealing the Unique Role of Water in the Formation of Benzothiazoles: an Experimental and Computational Study.

We present here a joint experimental and computational study on the formation of benzothiazoles. Our investigation reveals a green protocol for accessing benzothiazoles from acyl chlorides using either water alongside a reducing agent as the reaction medium or in combination with stoichiometric amounts of a weak acid, instead of the harsh conditions and catalysts previously reported. Specifically, we show that a protic solvent, particularly water, enables the formation of 2-substituted benzothiazoles from N-acyl 1,2-aminothiophenols already at room temperature, without the need for strong acids or metal catalysts. DFT Molecular Dynamics simulations coupled with advanced enhanced sampling techniques provide a clear understanding of the catalytic role of water. We demonstrate how bulk water - due to its extended network of hydrogen bonds and an efficient Grotthuss mechanism - provides a reaction path that strongly reduces the reaction barriers compared to aprotic environments, namely more than 80 kJ/mol for the first reaction step and 250 kJ/mol for the second. Finally, we discuss the influence of different aliphatic and aromatic substituents with varying electronic properties on chemical reactivity. Besides providing in-depth mechanistic insights, we believe that our findings pave the way for a greener route toward an important class of bioactive molecules.

Unveiling the Antifouling Potential of Stabilized Poly(phosphorus ylides).

Zwitterionic polymers have emerged as highly attractive building blocks for antifouling coatings in biomedical applications. Notably, these polymers offer effective alternatives to the widely used poly(ethylene glycol) (PEG), which has raised concerns regarding its immunotoxicity and the development of PEG-specific antibodies. Polymeric ylides, a largely overlooked class of zwitterionic polymers, have been reported as effective antifouling scaffolds. However, the reported subclasses, poly(sulfur ylides) and N-oxides, lack structural diversity and chemical variability. In this study, we present the synthesis and characterization of polymeric phosphorus ylides as an unexplored class of poly(ylides) with significantly increased structural diversity, which is of high value when designing future ylide-based antifouling materials. Our findings demonstrate that, owing to their low dipole moments and hydration layers, these polymeric phosphorus ylides significantly reduce bacterial attachment. Furthermore, we observe selective toxicity toward bacteria rather than mammalian cells. The bactericidal nature of poly(phosphorus ylides), coupled with their expanded chemical space, provides a distinct advantage over existing materials, including zwitterionic polymers from betaine scaffolds. We anticipate that these unexplored structures will broaden the scope of antifouling applications for poly(ylides).

Click'n lock: rapid exchange between unsymmetric tetrazines and thiols for reversible, chemoselective functionalisation of biomolecules with on-demand bioorthogonal locking.

The late-stage functionalisation and diversification of complex structures including biomolecules is often achieved with the help of click chemistry. Besides employing irreversible click-like reactions, many synthetic applications benefit from reversible click reaction strategies, so called de-/trans-click approaches. Yet, the combination of both, reversible and irreversible click chemistry - while still respecting the stringent criteria of click transformations - remains so far elusive for modifications of biomolecular structures. Here, we report click'n lock as a concept that enables reversible click reactions and on-demand locking of chemical entities, thus switching from reversible to irreversible modifications of complex biomolecules. For this purpose, we employ the tetrazine-thiol exchange (TeTEx) reaction as a fully traceless click reaction with second order rate constants k2 higher than 2 M-1 s-1 within aqueous environments. Employing TeTEx as a reversible click reaction for the chemoselective modification of biomolecules is made possible by the use of 3,6-disubstituted 1,2,4,5-tetrazines bearing a single sulfide residue. The inherent reactivity of tetrazines towards inverse electron demand Diels-Alder (IEDDA) reactions allows to stabilize the clicked structure, switching from reversible to irreversible systems (click'n lock).

Zwitterionic Polymeric Sulfur Ylides with Minimal Charge Separation Open a New Generation of Antifouling and Bactericidal Materials.

Zwitterionic polymers are widely employed hydrophilic building blocks for antifouling coatings with numerous applications across a wide range of fields, including but not limited to biomedical science, drug delivery and nanotechnology. Zwitterionic polymers are considered as an attractive alternative to polyethylene glycol because of their biocompatibility and effectiveness to prevent formation of biofilms. To this end, zwitterionic polymers are classified in two categories, namely polybetaines and polyampholytes. Yet, despite a fundamental interest to drive the development of new antifouling materials, the chemical composition of zwitterionic polymer remains severely limited. Here, we show that poly(sulfur ylides) that belong to the largely overlooked class of poly(ylides), effectively prevent the formation of biofilms from pathogenic bacteria. While surface energy analysis reveals strong hydrogen-bond acceptor capabilities of poly(sulfur ylide), membrane damage of pathogenic bacteria induced by poly(sulfur ylides) indicates toxicity towards bacteria while not affecting eucaryotic cells. Such synergistic effect of poly(sulfur ylides) offers distinct advantages over polyethylene glycol when designing new antifouling materials. We expect that our findings will pave the way for the development of a range of ylide-based materials with antifouling properties that have yet to be explored, opening up new directions at the interface of chemistry, biology, and material science.

Synthesis of multi-module low density lipoprotein receptor class A domains with acid labile cyanopyridiniumylides (CyPY) as aspartic acid masking groups.

Low-density lipoprotein receptor class A domains (LA modules) are common ligand-binding domains of transmembrane receptors of approximately 40 amino acids that are involved in several cellular processes including endocytosis of extracellular targets. Due to their wide-ranging function and distribution among different transmembrane receptors, LA modules are of high interest for therapeutic applications. However, the efficient chemical synthesis of LA modules and derivatives is hindered by complications, many arising from the high abundance of aspartic acid and consequent aspartimide formation. Here, we report a robust, efficient and general applicable chemical synthesis route for accessing such LA modules, demonstrated by the synthesis and folding of the LA3 and LA4 modules of the low-density lipoprotein receptor, as well as a heterodimeric LA3-LA4 constructed by chemical ligation. The synthesis of the aspartic acid-rich LA domain peptides is made possible by the use of cyanopyridiniumylides (CyPY) - reported here for the first time - as a masking group for carboxylic acids. We show that cyanopyridiniumylide masked aspartic acid monomers are readily available building blocks for solid phase peptide synthesis and successfully suppress aspartimide formation. Unlike previously reported ylide-based carboxylic acid protecting groups, CyPY protected aspartic acids are converted to the free carboxylic acid by acidic hydrolysis and are compatible with all common residues and protecting groups. The chemical synthesis of Cys- and Asp-rich LA modules enables new access to a class of difficult to provide, but promising protein domains.

Reaching new lights: a review on photo-controlled nanomedicines and their in vivo evaluation.

The selective and efficient delivery of bioactive molecules to sites of interest remains a formidable challenge in medicine. In recent years, it has been shown that stimuli-responsive drug delivery systems display several advantages over traditional drug administration such as an improved pharmacokinetic profile and the desirable ability to gain control over release. Light emerged as one of the most powerful stimuli due to its high biocompatibility, spatio-temporal control, and non-invasiveness. On the road to clinical translation, various chemical systems of high complexity have been reported with the aim to improve efficacy, safety, and versatility of drug delivery under complex biological conditions. For future research on the chemical design of such photo-controlled nanomedicines, it is essential to gain an understanding of their in vivo translation and efficiency. Here, we discuss photo-controlled nanomedicines that have been evaluated in vivo and provide an overview of the state-of-the-art that should guide future research design.

The effect of resource availability on interspecific competition between a native and an invasive ant.

Interspecific competition influences the composition of ecological communities. Species may differ in their needs for different resources, therefore resource availability may determine the outcome of interspecific interactions. Species often compete over food, shelter or both. When more than one resource is limited, different species may prioritize different resources. To determine the impact of resource availability on the competitive relationship between an invasive and a native species, we examined interactions between groups of the invasive Argentine ant (Linepithema humile) and the native odorous ant (Tapinoma sessile) over (1) food, (2) shelter or (3) both simultaneously. We further examined the mechanisms underlying the competitive relationship, asking whether aggressive interactions, exploratory behaviour or the order of arrival at a resource explained resource use. Shelter was preferred by both species when no competitors were present. In a competitive setting, L. humile groups controlled shelter through aggressive displacement but lost control over food due to investment of workers in the control of shelter. Thus, there are tradeoffs when competing over multiple resources and aggressive interactions allow invasive species to displace native species from a preferred resource. This article is part of the theme issue 'Intergroup conflict across taxa'.

Switching on prodrugs using radiotherapy.

Chemotherapy is a powerful tool in the armoury against cancer, but it is fraught with problems due to its global systemic toxicity. Here we report the proof of concept of a chemistry-based strategy, whereby gamma/X-ray irradiation mediates the activation of a cancer prodrug, thereby enabling simultaneous chemo-radiotherapy with radiotherapy locally activating a prodrug. In an initial demonstration, we show the activation of a fluorescent probe using this approach. Expanding on this, we show how sulfonyl azide- and phenyl azide-caged prodrugs of pazopanib and doxorubicin can be liberated using clinically relevant doses of ionizing radiation. This strategy is different to conventional chemo-radiotherapy radiation, where chemo-sensitization of the cancer takes place so that subsequent radiotherapy is more effective. This approach could enable site-directed chemotherapy, rather than systemic chemotherapy, with 'real time' drug decaging at the tumour site. As such, it opens up a new era in targeted and directed chemotherapy.

Synthesis of Polymers Containing Potassium Acyltrifluoroborates (KATs) and Post-polymerization Ligation and Conjugation.

We report the synthesis of monomers for atom-transfer radical polymerization (ATRP) and a reversible addition-fragmentation chain transfer (RAFT) agent bearing trifluoroborate iminiums (TIMs), which are quantitatively converted into potassium acyltrifluoroborates (KATs) after polymerization. The resulting KAT-containing polymers are suitable for rapid amide-forming ligations for both post-polymerization modification and polymer conjugation. The polymer conjugation occurs rapidly, even under dilute (micromolar) aqueous conditions at ambient temperatures, thereby enabling the synthesis of a variety of linear and star-shaped block copolymers. In addition, we applied post-polymerization modification to the covalent linking of a photocaged cyclic antibiotic (gramicidin S) to the side chains of the KAT-containing copolymer. Cellular assays revealed that the polymer-antibiotic conjugate is biocompatible and provides efficient light-controlled release of the antibiotic on demand.

Prevention of aspartimide formation during peptide synthesis using cyanosulfurylides as carboxylic acid-protecting groups.

Although peptide chemistry has made great progress, the frequent occurrence of aspartimide formation during peptide synthesis remains a formidable challenge. Aspartimide formation leads to low yields in addition to costly purification or even inaccessible peptide sequences. Here, we report an alternative approach to address this longstanding challenge of peptide synthesis by utilizing cyanosulfurylides to mask carboxylic acids by a stable C-C bond. These functional groups-formally zwitterionic species-are exceptionally stable to all common manipulations and impart improved solubility during synthesis. Deprotection is readily and rapidly achieved under aqueous conditions with electrophilic halogenating agents via a highly selective C-C bond cleavage reaction. This protecting group is employed for the synthesis of a range of peptides and proteins including teduglutide, ubiquitin, and the low-density lipoprotein class A. This protecting group strategy has the potential to overcome one of the most difficult aspects of modern peptide chemistry.

Novel copolymers drive differentiation of human adipose derived stem cells towards chondrocytes and osteoblasts identified by high-throughput approach.

Human adipose derived stem cells (hASCs) were seeded onto polymer microarrays that had been fabricated using a variety of acrylate monomers to discover novel substrates that induced differentiation towards chondrocytes and osteoblasts. Flow cytometric analysis showed that both CD105 and CD49d positive hASCs increased rapidly with passage number on the lead polymers, while quantitative PCR analysis showed that the substrate synthesized from methacryloxyethyltrimethyl ammonium chloride, N,N-diethylaminoethyl methacrylate and cyclohexyl methacrylate enhanced chondrogenesis and osteogenensis some 4 and 25 times respectively in terms of the expression of SOX9 and ALP in differentiated stem cells. These copolymers substrates thus have great potential for application in the purification, generation and expansion of defined hASC's and the controlled differentiation of of cells for possible clinical application.

Collective responses to heterospecifics emerge from individual differences in aggression.

Variation in individual behavior among group members impacts collective outcomes. The ability of both individuals and groups to outcompete others can determine access to resources. The invasive Argentine ant, Linepithema humile, dominates resources and displaces native species. To determine how access to resources by groups of L. humile is impacted by their behavioral composition, we first determined that L. humile workers consistently vary in aggressive behavior. We then asked if variation in aggression within a group influences the group's ability to access a resource in the presence of cues of a native species, Tapinoma sessile. We found that the behavioral composition of L. humile groups impacted the groups' collective response to cues of T. sessile. Group behavior was the result of mostly additive, rather than synergistic, combinations of the behaviors of the group members. The behavior of groups that contained 50% highly aggressive and 50% low-aggression individuals was similar to the average of the behaviors of groups of all highly aggressive and groups of all low-aggression individuals. Uncovering the mechanisms that allow social invasive species to dominate the ecological communities they invade can inform the mitigation of invasion.

The Emerging Role of Tetrazines in Drug-Activation Chemistries.

Traditionally, prodrug activation has been limited to enzymatic triggers or gross physiological aberrations, such as pH, that offer low selectivity and control over dosage. In recent years, the field of prodrug activation chemistry has been transformed by the use of bioorthogonal reactions that can be carried out under biological conditions at sub-millimolar concentrations, with the tetrazine-mediated inverse electron demand Diels-Alder reaction amongst the most recognised. Their high reaction rates, chemoselectivity and excellent biocompatibility make tetrazines ideal small molecules for activating prodrugs. Recently the tetrazine moiety has been used as a prodrug for a pyridazine thus broadening the scope of prodrug systems. This article discusses the concept of using tetrazines as small-molecule activators for prodrugs, and provides an overview of tetrazine-based prodrug systems, with a particular focus on the recently reported prodrug-prodrug activation strategy.

Tetrazine-mediated bioorthogonal prodrug-prodrug activation.

The selective and biocompatible activation of prodrugs within complex biological systems remains a key challenge in medical chemistry and chemical biology. Herein we report, for the first time, a dual prodrug activation strategy that fully satisfies the principle of bioorthogonality by the symbiotic formation of two active drugs. This dual and traceless prodrug activation strategy takes advantage of the INVDA chemistry of tetrazines (here a prodrug), generating a pyridazine-based miR21 inhibitor and the anti-cancer drug camptothecin and offers a new concept in prodrug activation.

Synthesis and characterization of biodegradable poly(ether-ester) urethane acrylates for controlled drug release.

Three polyether-ester triblock diols, with various molecular weights, were synthesized from ε-caprolactone and polyethylene glycol and used, with diisocyanates, as soft segments for the preparation of polyurethane acrylate oligomers. The polyurethane acrylates were used to generate cross-linked polyurethane films via UV initiated polymerization with and without cargo incorporation. Degradation experiment indicated that in PBS/H2O2/CoCl2 the films degraded rapidly compared to PBS alone or with lipase. The polyurethane membrane loaded with the antibiotic tetracycline, demonstrated prolonged release over 200h, suggesting that the polymers could be used as an implant coating for controlled drug release.

Tetrazine-Responsive Self-immolative Linkers.

Molecules that undergo activation or modulation following the addition of benign external small-molecule chemical stimuli have numerous applications. Here, we report the highly efficient "decaging" of a variety of moieties by activation of a "self-immolative" linker, by application of water-soluble and stable tetrazine, including the controlled delivery of doxorubicin in a cellular context.

Tuning the emission properties of a fluorescent polymer using a polymer microarray approach - identification of an optothermo responsive polymer.

Polymer microarrays were prepared using inkjet printing mixtures of acrylate monomers each with a common fluorescent fluorene co-polymer. Fluorescence analysis of each of the features on the array allowed identification of polymers that could tune the fluorescence under a variety of insults. The "hit" polymers were made into beads via reverse suspension polymerization and their fluorescence properties were analyzed.

Nanoparticle "switch-on" by tetrazine triggering.

This work describes how a small-molecule chemical trigger, reacting through the mediatory of an inverse electron demand Diels-Alder reaction, results in enhanced cellular uptake and selective nanoparticle disintegration and cargo liberation, via gross polymeric morphological alterations. The power of these responsive nanoparticles is demonstrated through encapsulation of the anti-cancer agent doxorubicin and its triggered release, allowing controlled cell death in response to a small-molecule chemical trigger.