Additive-assisted macroscopic self-assembly and control of the shape of assemblies based on host-guest interaction.

In these decades, considerable attention has focused on supramolecular polymers due to their unique structures and properties. More recently, macroscopic supramolecular polymers have attracted increasing interest from not only biologists but also materials scientists inspired by the sophisticated structures and functions of living organisms. Since the functions of supramolecular polymers are strongly dependent on their shape, control of the shape is an important issue in controlling the functions of supramolecular polymers. However, the control of shape in macroscopic supramolecular assemblies has not yet been sufficiently investigated. Previously, we studied the macroscopic self-assembly behavior of super absorbent polymer (SAP) microparticles modified with ß-cyclodextrin (ßCD) and adamantane (Ad) residues (ßCD(x)-SAP and Ad(y)-SAP microparticles, where x and y are the mol% contents of ßCD and Ad residues, respectively). More elongated assemblies were formed at higher y, indicating that the shape of assemblies can be controlled by varying the interaction strength. The noteworthy is that 1-adamantanamine hydrochloride (AdNH3Cl) assisted the formation of assemblies from ßCD(x)-SAP and Ad(y)-SAP microparticles, indicating that AdNH3Cl acts as a chemical stimulus for macroscopic assemblies of ßCD(x)-SAP and Ad(y)-SAP microparticles. In this study, we have thus studied the assembling behavior of ßCD(x)-SAP microparticles with Ad(y)-SAP microparticles and unmodified SAP microparticles assisted by AdNH3Cl, as well as the shape of the resulting macroscopic assemblies. AdNH3Cl assisted the formation of assemblies from ßCD(16.2)-SAP and Ad(15.1)-SAP microparticles, in which AdNH3Cl crosslinked the SAP microparticles through the formation of inclusion complexes of ßCD residues with the Ad residue and the electrostatic interaction of ammonium and carboxylate residues. Assemblies of ßCD(26.7)-SAP and unmodified SAP microparticles were formed at the concentrations of AdNH3Cl ([AdNH3Cl]0) higher than a certain level (ca. 0.05 mM). The aspect ratio (a/b) of assemblies showed a maximum at [AdNH3Cl]0 ~ 0.10 mM, indicating that the chemical stimulus, i.e., addition of AdNH3Cl, controls the shape of assemblies formed from ßCD(26.7)-SAP and unmodified SAP microparticles. This study suggests that other stimuli, e.g., heat, pH, light, redox, and force, can be utilized to control the shape of macroscopic assemblies based on supramolecular interactions.

Interaction of Cyclodextrins with Amphiphilic Alternating Cooligomers Possessing the Dense Triazole Backbone.

The interaction of cyclodextrins (CDs) with structure-controlled polymers is expected to provide significant insights into macromolecular recognition. However, the interaction of CDs with structure-controlled polymers has been an underexamined issue of investigation. Herein, alternating amphiphilic cooligomers (oligoCnAH, where n denotes the carbon number of alkyl groups; n = 4, 8, and 12) were synthesized by copper(I)-catalyzed azide-alkyne cycloaddition polymerization of heterodimers of 4-azido-5-hexynoic acid (AH) derivatives carrying N-alkylamide and t-butyl (tBu) ester side chains, followed by hydrolysis of the tBu ester, to study the interaction of CDs with oligoCnAH by 1H NMR, nuclear Overhauser effect spectroscopy, and pulse-field-gradient spin-echo NMR. These NMR studies indicated that αCD interacted with oligoC4AH, αCD and ßCD interacted with oligoC8AH, and all CDs interacted with oligoC12AH. Based on the equilibrium models proposed, the binding constants were evaluated for the binary mixtures, which showed interaction. Comparing the interactions of the CDs/oligoC12AH binary mixtures with those of the binary mixtures of CDs and alternating copolymers of sodium maleate and dodecyl vinyl ether (polyC12M), it is concluded that oligoC12AH forms less stable micelles than does polyC12M presumably because of the lower molecular weight, the hydrophilic amide groups in the side chain, and the longer interval between neighboring C12 groups in oligoC12AH.

Synthesis of Dense 1,2,3-Triazole Oligomers Consisting Preferentially of 1,5-Disubstituted Units via Ruthenium(II)-Catalyzed Azide-Alkyne Cycloaddition.

Ruthenium(II)-catalyzed azide-alkyne cycloaddition (RuAAC) polymerization of t-butyl 4-azido-5-hexynoate (tBuAH), i.e., a heterobifunctional monomer carrying azide and alkyne moieties, was investigated in this study. RuAAC of the monofunctional precursors of tBuAH yielded a dimer possessing a 1,5-disubstituted 1,2,3-triazole moiety. 1H NMR data showed that the dimer was a mixture of diastereomers. Polymerization of tBuAH using ruthenium(II) (Ru(II)) catalysts produced oligomers of Mw ≈ (2.7-3.6) × 103 consisting of 1,5-disubstituted 1,2,3-triazole units (1,5-units) as well as 1,4-disubstituted 1,2,3-triazole units (1,4-units). The fractions of 1,5-unit (f1,5) were roughly estimated to be ca. 0.8 by comparison of signals of the methine and triazole protons in 1H NMR spectra, indicating that RuAAC proceeded preferentially and thermal Huisgen cycloaddition (HC) somehow took place during the polymerization. The oligomer samples obtained were also characterized by solubility test, size exclusion chromatography (SEC), ultraviolet-visible (UV-Vis) absorption spectroscopy, and thermogravimetric analysis (TGA). The UV-Vis and TGA data indicated that the oligomer samples contained a substantial amount of Ru(II) catalysts. To the best of our knowledge, this is the first report on dense 1,2,3-triazole oligomers consisting of 1,5-units linked via a carbon atom.

Preferential formation of specific hexose and heptose in the formose reaction under microwave irradiation.

To realize sustainable societies, the production of organic compounds not based on fossil resources should be developed. Thus, C1 chemistry, utilizing one-carbon compounds as starting materials, has been of increasing importance. In particular, the formose reaction is promising because the reaction produces sugars (monosaccharides) from formaldehyde under basic conditions. On the other hand, since microwave (MW) induces the rotational motion of molecules, MW irradiation often improves the selectivity and efficiency of reactions. In this study, the formose reaction under MW irradiation was thus investigated under various conditions. The formose reaction proceeded very fast using 1.0 mol per kg formaldehyde and 55 mmol per kg calcium hydroxide (Ca(OH)2) as a catalyst at a high set temperature (150 °C) for a short time (1 min) to form preferentially specific hexose and heptose. The major products were isolated by thin layer chromatography and characterized by mass spectroscopy and NMR. These characterization data elucidated that the hexose and heptose were 2-hydroxymethyl-1,2,4,5-tetrahydroxy-3-pentanone (C6*) and 2,4-bis(hydroxymethyl)-1,2,4,5-tetrahydroxy-3-pentanone (C7*), respectively. On the basis of these observations, as well as density functional theory calculations, a plausible reaction pathway was also discussed; once 1,3-dihydroxyacetone is formed, consecutive aldol reactions favorably occur to form C6* and C7*.

Synthesis of New Thermoresponsive Polymers Possessing the Dense 1,2,3-Triazole Backbone.

Thermoresponsive water-soluble polymers, aqueous solutions of which undergo lower critical solution temperature (LCST)-type phase separation, have been investigated in detail for several decades. To develop LCST-type thermoresponsive polymers with new polymer backbone, 4-azido-5-hexynamide (AHA) derivatives were designed as monomers for copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) polymerization. AHA derivatives carrying secondary amide side chains, that is, 4-azido-N-methyl-5-hexynamide (M), 4-azido-N-ethyl-5-hexynamide (E), and 4-azido-N-isopropyl-5-hexynamide (iP), were first synthesized and polymerized by CuAAC to obtain polymers (poly(M), poly(E), and poly(iP)). Contrary to our expectation, poly(M), poly(E), and poly(iP) were insoluble in water and many organic solvents presumably because of the formation of hydrogen bonding between the amide side chains or between the amide side chains and triazole residues in the backbone. Thus, AHA derivatives carrying tertiary amide side chains, that is, 4-azido-N,N-dimethyl-5-hexynamide (MM), 4-azido-N-ethyl-N-methyl-5-hexynamide (ME), 4-azido-N-isopropyl-N-methyl-5-hexynamide (MiP), and 4-azido-N,N-diethyl-5-hexynamide (EE), were also synthesized and polymerized to yield polymers (poly(MM), poly(ME), poly(MiP), and poly(EE)). These polymers were soluble in a number of common organic solvents. It is noteworthy that poly(MM) and poly(ME) were also soluble in water. The phase separation behavior of 1.0 wt % aqueous solutions of poly(MM) and poly(ME) was then investigated by transmittance measurements. These data indicated that poly(ME) was an LCST-type thermoresponsive polymer, whereas poly(MM) was not. A large hysteresis was observed in the transmittance measurements for the poly(ME) aqueous solution because of slow rehydration after phase separation. The phase separation behavior was investigated preliminarily by differential scanning calorimetry and 1H NMR.

Synthetic hydrogel nanoparticles for sepsis therapy.

Sepsis is a life-threatening condition caused by the extreme release of inflammatory mediators into the blood in response to infection (e.g., bacterial infection, COVID-19), resulting in the dysfunction of multiple organs. Currently, there is no direct treatment for sepsis. Here we report an abiotic hydrogel nanoparticle (HNP) as a potential therapeutic agent for late-stage sepsis. The HNP captures and neutralizes all variants of histones, a major inflammatory mediator released during sepsis. The highly optimized HNP has high capacity and long-term circulation capability for the selective sequestration and neutralization of histones. Intravenous injection of the HNP protects mice against a lethal dose of histones through the inhibition of platelet aggregation and migration into the lungs. In vivo administration in murine sepsis model mice results in near complete survival. These results establish the potential for synthetic, nonbiological polymer hydrogel sequestrants as a new intervention strategy for sepsis therapy and adds to our understanding of the importance of histones to this condition.

Synthesis of a New Polyanion Possessing Dense 1,2,3-Triazole Backbone.

Polyanions are an important class of water-soluble polymers because polyanions are utilized in a wide range of industrial fields. It is thus a great challenge to develop polyanions with novel structures to make their applications broader. In this study, a new polyanion with a dense 1,2,3-triazole backbone, poly(4-azido-5-hexanoic acid) (poly(AH)), was synthesized by copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) polymerization of t-butyl 4-azido-5-hexanoate followed by hydrolysis of the t-butyl ester groups. Turbidimetric and potentiometric titration data indicated that poly(AH) was well soluble in water under basic conditions (pH < 7) and a weaker polyanion (apparent pKa = 5.4) than polyacrylic acid (apparent pKa = 4.5). Adsorption tests exhibited that sodium salt of poly(AH) (poly(AH)Na) adsorbed most preferably Fe3+ among the four metal ions examined, i.e., Cu2+, Pb2+, Li+, and Fe3+. 1H spin-lattice relaxation time measurements indicated that Fe3+ ions were adsorbed favorably onto the 1,2,3-triazole residues.

Synthesis of Dense 1,2,3-Triazole Polymers Soluble in Common Organic Solvents.

Aiming at synthesis of dense 1,2,3-triazole polymers soluble in common organic solvents, a new 3-azido-1-propyne derivative, i.e., t-butyl 4-azido-5-hexynoate (tBuAH), was synthesized and polymerized by copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) and Huisgen cycloaddition (HC). CuAAC polymerization produced poly(tBuAH) composed of 1,4-disubstituted 1,2,3-triazole units (1,4-units), whereas HC polymerization gave poly(tBuAH) composed of 1,4- and 1,5-disubstituted 1,2,3-triazole units (1,4- and 1,5-units). In HC polymerization, the fraction of 1,4-unit (f1,4) decreased with the permittivity of solvent used. Differential scanning calorimetry data indicated that the melting point of poly(tBuAH) increased from 61 to 89 °C with increasing f1,4 from 0.38 to 1.0, indicative of higher crystallinity of poly(tBuAH) composed of 1,4-unit. Preliminary steady-state fluorescence study indicated that all the poly(tBuAH) samples of different f1,4 emitted weak but significant fluorescence in DMF. The maximum of fluorescence band shifted from ca. 350 to ca. 450 nm with varying the excitation wavelength from 300 to 400 nm.

The macroscopic shape of assemblies formed from microparticles based on host-guest interaction dependent on the guest content.

Biological macroscopic assemblies have inspired researchers to utilize molecular recognition to develop smart materials in these decades. Recently, macroscopic self-assemblies based on molecular recognition have been realized using millimeter-scale hydrogel pieces possessing molecular recognition moieties. During the study on macroscopic self-assembly based on molecular recognition, we noticed that the shape of assemblies might be dependent on the host-guest pair. In this study, we were thus motivated to study the macroscopic shape of assemblies formed through host-guest interaction. We modified crosslinked poly(sodium acrylate) microparticles, i.e., superabsorbent polymer (SAP) microparticles, with ß-cyclodextrin (ßCD) and adamantyl (Ad) residues (ßCD(x)-SAP and Ad(y)-SAP microparticles, respectively, where x and y denote the mol% contents of ßCD and Ad residues). Then, we studied the self-assembly behavior of ßCD(x)-SAP and Ad(y)-SAP microparticles through the complexation of ßCD with Ad residues. There was a threshold of the ßCD content in ßCD(x)-SAP microparticles for assembly formation between x = 22.3 and 26.7. On the other hand, the shape of assemblies was dependent on the Ad content, y; More elongated assemblies were formed at a higher y. This may be because, at a higher y, small clusters formed in an early stage can stick together even upon collisions at a single contact point to form elongated aggregates, whereas, at a smaller y, small clusters stick together only upon collisions at multiple contact points to give rather circular assemblies. On the basis of these observations, the shape of assembly formed from microparticles can be controlled by varying y.

Signalling molecular recognition nanocavities with multiple functional groups prepared by molecular imprinting and sequential post-imprinting modifications for prostate cancer biomarker glycoprotein detection.

Fluorescent-signalling molecularly-imprinted nanocavities possessing orthogonal dual interaction sites for the detection of prostate cancer biomarker glycoprotein were constructed through molecular imprinting and sequential multistep post-imprinting modifications (PIMs) using a newly designed multi-functionalised PIM reagent (PIR). The PIR, possessing an interaction site and dual reaction sites for PIMs, enabled us to introduce multiple functions including interaction sites and fluorescent reporter groups in a single PIM site, leading to the sensitive fluorescent detection of target glycoproteins with a high signal-to-noise ratio. Prostate specific antigen (PSA), used as a biomarker for prostate-related diseases, was selected as a target glycoprotein. Surface-initiated atom transfer radical polymerisation from template PSA immobilised the substrate with a functional monomer possessing a phenyl boronic acid group, where the template PSA was designed to possess polymerisation groups aligned with disulphide linkage. Using the thiol groups left after removing templates, PIR could be introduced as the 1st PIM. An evaluation of the effect of crosslinking density and blocking treatment on selective detection indicated that highly selective and sensitive detection of PSA was achieved. Furthermore, the 2nd PIM to introduce fluorescent molecules into the nanocavities led to the fluorescent detection of PSA. The new sequential PIM strategy using multi-functional PIR can potentially create various sophisticated artificial molecular recognition materials.

Molecularly Imprinted Nanocavities Capable of Ligand-Binding Domain and Size/Shape Recognition for Selective Discrimination of Vascular Endothelial Growth Factor Isoforms.

Vascular endothelial growth factor 165 (VEGF165) is known to be predominantly expressed in the first stage of vascularization; therefore, the detection of VEGF165 is important in the stage diagnosis of cancers. Molecularly imprinted nanocavities, capable of the selective discrimination of VEGF165 from other VEGF isoforms, were prepared by surface-initiated atom transfer radical polymerization. VEGF165 was immobilized on a gold-coated glass substrate by anchored heparin moieties, where the immobilized heparin was able to capture VEGF165 by binding with the heparin-binding domain (HBD) on VEGF165. Molecular imprinting was conducted on the immobilized VEGF165 by using methacrylic acid (MAA) as a functional monomer to interact with basic amino acids outside of the HBD of VEGF165 by electrostatic interaction. After the removal of VEGF165 from the obtained polymer thin layer (ca. 7 nm), VEGF165-imprinted nanocavities remained, in which the heparin moiety and MAA residues were located in suitable positions for VEGF165 recognition. The molecularly imprinted polymer (MIP) thin layer showed a binding affinity for VEGF165 (dissociation constant: 3.4 nM) that was ten times higher than that of the substrate before polymerization (heparin-immobilized substrate). A much lower binding affinity for VEGF121, which contains no heparin-binding domain, was observed. Moreover, the MIP thin layer distinguished VEGF165 from VEGF189, which possesses a larger molecular size than VEGF165, an amino acid sequence homology of 87%, and contains HBDs, whereas the heparin-immobilized substrate showed almost no selectivity. These results suggested that the heparin moiety within the nanocavity provided HBD selectivity and the polymer matrix composed of the molecularly imprinted nanocavity provided size/shape selectivity, which resulted in the highly selective discrimination of VEGF isoforms.

Oriented, molecularly imprinted cavities with dual binding sites for highly sensitive and selective recognition of cortisol.

Novel, molecularly imprinted polymers (MIPs) were developed for the highly sensitive and selective recognition of the stress marker cortisol. Oriented, homogeneous cavities with two binding sites for cortisol were fabricated by surface-initiated atom transfer radical polymerization, using a cortisol motif template molecule (TM1) which consists of a polymerizable moiety attached at the 3-carbonyl group of cortisol via an oxime linkage and an adamantane carboxylate moiety coupled with the 21-hydroxyl group. TM1 was orientationally immobilized on a ß-cyclodextrin (ß-CD)-grafted gold-coated sensor chip by inclusion of the adamantane moiety of TM1, followed by copolymerization of a hydrophilic comonomer, 2-methacryloyloxyethyl phosphorylcholine, with or without a cross-linker, N,N'-methylenebisacrylamide. Subsequent cleavage of the oxime linkage leaves the imprinted cavities that contain dual binding sites-namely, the aminooxy group and ß-CD-capable of oxime formation and hydrophobic interaction, respectively. As an application, MIP-based picomolar level detection of cortisol was demonstrated by a competitive binding assay using a fluorescent competitor. Cross-linking of the MIP imparts rigidity to the binding cavities, and improves the selectivity and sensitivity significantly, reducing the limit of detection to 4.8 pM. In addition, detection of cortisol in saliva samples was demonstrated as a feasibility study.

Synthesis of grafted phosphorylcholine polymer layers as specific recognition ligands for C-reactive protein focused on grafting density and thickness to achieve highly sensitive detection.

We studied the effects of layer thickness and grafting density of poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) thin layers as specific ligands for the highly sensitive binding of C-reactive protein (CRP). PMPC layer thickness was controlled by surface-initiated activators generated by electron transfer for atom transfer radical polymerization (AGET ATRP). PMPC grafting density was controlled by utilizing mixed self-assembled monolayers with different incorporation ratios of the bis[2-(2-bromoisobutyryloxy)undecyl] disulfide ATRP initiator, as modulated by altering the feed molar ratio with (11-mercaptoundecyl)tetra(ethylene glycol). X-ray photoelectron spectroscopy and ellipsometry measurements were used to characterize the modified surfaces. PMPC grafting densities were estimated from polymer thickness and the molecular weight obtained from sacrificial initiator during surface-initiated AGET ATRP. The effects of thickness and grafting density of the obtained PMPC layers on CRP binding performance were investigated using surface plasmon resonance employing a 10 mM Tris-HCl running buffer containing 140 mM NaCl and 2 mM CaCl2 (pH 7.4). Furthermore, the non-specific binding properties of the obtained layers were investigated using human serum albumin (HSA) as a reference protein. The PMPC layer which has 4.6 nm of thickness and 1.27 chains per nm(2) of grafting density showed highly sensitive CRP detection (limit of detection: 4.4 ng mL(-1)) with low non-specific HSA adsorption, which was improved 10 times than our previous report of 50 ng mL(-1).