橋本 剛

Developing biocompatible nitric oxide (NO) photoreleasing nanoconstucts is of great interest in view of the large variety of biological roles that NO plays and the unique advantage light offers in controlling NO release in space and time. In this contribution, we report the supramolecular assemblies of two NO photodonors (NOPDs), NBF-NO and RHD-NO, as water-dispersible nanogels, ca. 10 nm in diameter, based on γ-cyclodextrins (γ-CDng). These NOPDs, containing amino-nitro-benzofurazan and rhodamine chromophores as light harvesting antennae, can be activated by visible light, are highly hydrophobic and can be effectively entrapped within the γ-CDng. Despite being confined in a very restricted environment, neither NOPD suffer self-aggregation and preserve their photochemical and photophysical properties well. The blue light excitation of the weakly fluorescent γ-CDng/NBF-NO complex results in effective NO release and the concomitant generation of the highly green, fluorescent co-product, which acts as an optical NO reporter. Moreover, the green light excitation of the persistent red fluorescent γ-CDng/RHD-NO triggers NO photorelease without significantly modifying the emission properties. The activatable and persistent fluorescence emissions of the NOPDs are useful for monitoring their interactions with the Gram-positive methicillin-resistant Staphylococcus aureus, whose growth is significantly inhibited by γ-CDng/RHD-NO upon green light irradiation.

ATP recognition has been achieved by exploiting the self-assembly of boronic acid-appended cyclodextrin, a fluorescent probe, and ATP through multiple interactions.

Ultrasmall cyclodextrin nanogels were prepared by an inverse emulsion method using a cationic surfactant. These nanogels provide a highly hydrophobic inner surface, allowing efficient solubilisation of hydrophobic compounds in water.

Specifically designed electrochemical sensors are standing out as alternatives to enzyme-based biosensors for the sensing of metabolites. In our previous works, we developed a new electrochemical assay based on cyclodextrin supramolecular complexes. A ferrocene moiety (Fc) was chemically modified by phenylboronic acid (4-Fc-PB) and combined with two different kinds of cyclodextrins (CDs): β-CD and β-CD modified by a dipicolylamine group (dpa-p-HB-β-CDs) for the sensing of fructose and adenosine-triphosphate (ATP), respectively. The aim of the present work is to better comprehend the features underlining the aforementioned complex formation. For the first time, a study about inclusion phenomena between the 4-Fc-PB electroactive probe with β-CD and with dpa-p-HB-β-CD was performed by using nuclear magnetic resonance (NMR) analysis. In particular, we focused on providing insights on the interaction involved and on the calculation of the binding constant of 4-Fc-PB/β-CD supramolecular complex, and elucidation about a drift in the time observed during the control experiments of the electrochemical measurements for the 4-Fc-PB/dpa-p-HB-β-CD supramolecular complex. In this sense, this paper represents a step further in the explanation of the electrochemical results obtained, pointing out the nature of the interactions present both in the formation of the inclusions and in the sensing with the analytes.

We proposed an inclusion complex of γ-cyclodextrin with a benzoxaborole-based fluorescent probe as a highly sensitive and selective chemosensor for d-allulose.

We have developed a convenient and selective method for the detection of Gram-positive bacteria using a ditopic poly(amidoamine) (PAMAM) dendrimer probe. The dendrimer that was modified with dipicolylamine (dpa) and phenylboronic acid groups showed selectivity toward Staphylococcus aureus. The ditopic dendrimer system had higher sensitivity and better pH tolerance than the monotopic PAMAM dendrimer probe. We also investigated the mechanisms of various ditopic PAMAM dendrimer probes and found that the selectivity toward Gram-positive bacteria was dependent on a variety of interactions. Supramolecular interactions, such as electrostatic interaction and hydrophobic interaction, per se, did not contribute to the bacterial recognition ability, nor did they improve the selectivity of the ditopic dendrimer system. In contrast, the ditopic PAMAM dendrimer probe that had a phosphate-sensing dpa group and formed a chelate with metal ions showed improved selectivity toward S. aureus. The results suggested that the targeted ditopic PAMAM dendrimer probe showed selectivity toward Gram-positive bacteria. This study is expected to contribute to the elucidation of the interaction between synthetic molecules and bacterial surface. Moreover, our novel method showed potential for the rapid and species-specific recognition of various bacteria.

This study reports a novel, fast, easy, and sensitive detection method for bacteria which is urgently needed to diagnose infections in their early stages. Our work presents a complex of poly(amidoamine) dendrimer modified by phenylboronic acid and labeled by a fluorescent dansyl group (Dan-B8.5-PAMAM). Our system detects bacteria in 20 min with a sensitivity of approximately 104 colony-forming units (CFU)·mL−1. Moreover, it does not require any peculiar technical skills or expensive materials. The driving force for bacteria recognition is the binding between terminal phenylboronic acids on the probe and bacteria’s surface glycolipids, rather than electrostatic interactions. The aggregation caused by such binding reduces fluorescence. Even though our recognition method does not distinguish between live or dead bacteria, it shows selective antibacterial activity towards Gram-negative bacteria. This study may potentially contribute a new method for the convenient detection and killing of bacteria.

Cyclodextrins (CyDs) are water-soluble host molecules possessing a nanosized hydrophobic cavity. In the realm of molecular recognition, this cavity is used not only as a recognition site but also as a reaction medium, where a hydrophobic sensor recognizes a guest molecule. Based on the latter concept, we have designed a novel supramolecular sensing system composed of Zn(II)-dipicolylamine metal complex-based azobenzene (1-Zn) and 3A-amino-3A-deoxy-(2AS,3AS)-γ-cyclodextrin (3-NH2-γ-CyD) for sensing adenosine-5′-triphosphate (ATP). 1-Zn showed redshifts in the UV-Vis spectra and induced circular dichroism (ICD) only when both ATP and 3-NH2-γ-CyD were present. Calculations of equilibrium constants indicated that the amino group of 3-NH2-γ-CyD was involved in the formation of supramolecular 1-Zn/3-NH2-γ-CyD/ATP. The Job plot of the ICD spectral response revealed that the stoichiometry of 1-Zn/3-NH2-γ-CyD/ATP was 2:1:1. The pH effect was examined and 1-Zn/3-NH2-γ-CyD/ATP was most stable in the neutral condition. The NOESY spectrum suggested the localization of 1-Zn in the 3-NH2-γ-CyD cavity. Based on the obtained results, the metal coordination interaction of 1-Zn and the electrostatic interaction of 3-NH2-γ-CyD were found to take place for ATP recognition. The “reaction medium approach” enabled us to develop a supramolecular sensing system that undergoes multi-point interactions in water. This study is the first step in the design of a selective sensing system based on a good understanding of supramolecular structures.

Detection and visualization of phosphates such as ATP in living organisms can facilitate the elucidation of various biological events. Although substantial efforts had been made in this area, present methods have disadvantages such as the need for specialized equipment and poor sensitivities. To address these limitations, novel fluorescent probes, (di-(2-picolyl)amino)quinazolines, were developed for application in ATP detection. They selectively recognized copper ions by fluorescence quenching, and their copper complexes displayed fluorescence enhancement in the presence of phosphoric acid derivatives. This fluorescence on–off system enabled highly sensitive fluorescence detection of ATP when combined with a phenyl boronic acid-modified γ-cyclodextrin through a plausible multipoint recognition system.

糖分子やアデノシン三リン酸（ATP）といった生体関連分子をターゲットにした, より効率的なイオン・分子認識系の構築を目的として, 各種の超分子分子認識法を開発した。これらの系はジピコリルアミン－遷移金属錯体によるリン酸誘導体との結合と, フェニルボロン酸による糖のシスジオール基とのエステル形成をモチーフとして, 環状オリゴ糖であるシクロデキストリンの利用と分子間の弱い相互作用の組み合わせにより分子認識能を発現している。さらにはこれらの分子認識部を各種のナノ粒子に修飾することで, 蛍光あるいは電気化学的に細菌検出できる系も開発したので, それらについても報告する。

The construction of supramolecular recognition systems based on specific host-guest interactions has been studied in order to design selective chemical sensors. In this study, guest-responsive receptors for ATP have been designed with cyclodextrins (CyDs) as a basic prototype of the turn-on type fluorescent indicator. We synthesized dipicolylamine (DPA)-modified CyD-Cu2+ complexes (Cu1α, Cu1β, and Cu1γ), and evaluated their recognition capabilities toward phosphoric acid derivatives in water. The UV-Vis absorption and fluorescence spectra revealed that Cu1β selectively recognized ATP over other organic and inorganic phosphates, and that β-CyD had the most suitable cavity size for complexation with ATP. The 1D and 2D NMR analyses suggested that the ATP recognition was based on the host-guest interaction between the adenine moiety of ATP and the CyD cavity, as well as the recognition of phosphoric moieties by the Cu2+-DPA complex site. The specific interactions between the CyD cavity and the nucleobases enabled us to distinguish ATP from other nucleoside triphosphates, such as guanosine triphosphate (GTP), uridine triphosphate (UTP), and cytidine triphosphate (CTP). This study clarified the basic mechanisms of molecular recognition by modified CyDs, and suggested the potential for further application of CyDs in the design of highly selective supramolecular recognition systems for certain molecular targets in water.

In this study, we have developed a rational design strategy to obtain highly selective supramolecular recognition systems of cyclodextrins (CyDs) on the basis of the lock and key principle. We designed and synthesized dipicolylamine (dpa)-modified gamma-CyD Cu2+ complexes possessing an azobenzene unit (Cu center dot 1-gamma-CyD) and examined how they recognized phosphoric acid derivatives in water. The results revealed that Cu center dot 1-gamma-CyD recognized ATP with high selectivity over other phosphoric acid derivatives. The significant blue shift in the UV vis spectra and H-1 NMR analysis suggested that the selective ATP recognition was based on the multipoint interactions between the adenine moiety of ATP and both the CyD cavity and the azobenzene unit in addition to the recognition of phosphoric moieties by the Cu-dpa complex site. Our unique receptor made it capable of distinguishing ATP from AMP and ADP, revealing the discrimination of even a length of one phosphoric group. This study demonstrates that, compared to conventional recognition systems of CyDs, this multipoint recognition system confers a higher degree of selectivity for certain organic molecules, such as ATP, over their similar derivatives.

We report herein synthesis and characterization of four new organoruthenium(II) complexes of the type [RuH(CO)(PPh3)(2)(L-1,L-2)]Cl (1, 3) and [Ru(CO)(Cl)(2)(AsPh3)(L-1,L-2)] (2, 4) derived from the reaction of [RuHCl(CO)(EPh3)(3)] (E = P or As) with 2-(pyridine-2yl)benzoxazole (L-1) and 2-(pyridine-2yl)benzthiazole (L-2). Single-crystal X-ray diffraction data of 2 proved octahedral geometry of the complexes with a 1:1 ratio between the metal and the coordinated ligands. The binding affinities of 1-4 toward calf-thymus DNA (CT-DNA) and BSA were thoroughly studied by various spectroscopic techniques. Furthermore, the coordination compounds exhibit catecholase-like activities in the aerial oxidation of 3,5-di-tert-butylcatechol to the corresponding o-quinone and phosphatase-like activities in the hydrolysis of 4-nitrophenyl phosphate to 4-nitrophenolate ion. The kinetic parameters have been determined using Michaelis-Menten approach. The highest k(cat) values suggested that coordination compounds exhibit higher rates of catalytic efficacy.

We designed amphiphilic phenylboronic acid azoprobes (B-Azo-Cn) and evaluated their saccharide recognition function in relation to the micelle formation changes of the self-assembled B-Azo-Cn. First, we evaluated B-Azo-C8 in a 1% methanol-99% water solution under basic conditions. The wavelength of maximum absorption in the ultraviolet-visible (UV-vis) spectra of B-Azo-C8 was shifted, and the solution showed a color change with the addition of saccharides. The morphology of B-Azo-C8 was evaluated using dynamic light scattering (DLS) measurements and transmission electron microscopy (TEM) observations. B-Azo-C8 formed aggregates in the absence of saccharides and in the presence of glucose. In the presence of fructose, micelle-formed B-Azo-C8 was dispersed, indicating that B-Azo-C8 changed its dispersion state by recognizing fructose. The effect of alkyl chain length on the saccharide recognition ability was examined as well. B-Azo-C4 and B-Azo-C12 did not recognize saccharides in a 1% methanol-99% water solution under basic conditions, indicating that an appropriate alkyl chain length was required for recognizing saccharides. The control of the hydrophilic-lipophilic balance (HLB) was a key factor for saccharide recognition.

We have developed a new method for Staphylococcus aureus (S. aureus) detection, which employs fluorescent silica nanoparticles (FSiNPs) modified with metal-dipicolylamine complex (M-dpa-HCC). Among the M-dpa-HCC/FSiNP complexes, Cu-dpa-HCC/FSiNP formed large aggregates with S. aureus in 10 min, which were easily observed by the naked eye. The antibacterial activity of Cu-dpa-HCC/FSiNP was also confirmed.

<p>これまで量子スピンダイマーの研究は、低温で一重項状態にある系で、磁場・圧力等で、マグノンを誘起しそのBECを調べるものであった。本研究では、Ru二核金属錯体（β－ジケトナト）Ruダイマーの配位子を入れ替えることで、異種価数のキャリヤドープを行うことなく、Ru原子の価数をチューニングし、一重項状態を破壊することを目指す。講演ではNMRで見た磁性の変化にも言及する。</p>

The selective molecular recognition event by physical or chemical signals is a key concept for the design of novel supramolecular sensors. In this study, we designed a novel saccharide recognition system based on the self-assembly of phenylboronic acid azoprobes (1-BAzo-NPs) on the surface of the polyamidoamine (PAMAM) dendrimer in water. The two sulfonic acid moieties in 1-BAzo-NP enhanced the binding affinity of the azoprobe for the PAMAM dendrimer surface. UV-Vis spectral measurements indicated that 1-BAzo-NP showed poor saccharide recognition at pH 7.0, whereas the 1-BAzo-NP–PAMAM complex well recognized the saccharides, particularly glucose. The complexation with glucose yielded aggregates having diameters of 100–200 nm as determined by dynamic light scattering (DLS) measurements and transmission electron microscopy (TEM). The sensitivity to and selectivity for the saccharides were controlled by the density of the assembled phenylboronic acid azoprobes and PAMAM generation. Together, the results revealed that the selective saccharide recognition at pH 7.0 was feasible by boronic acid assembly on the PAMAM dendrimer surface.