Ryotaro Miki

Colorimetric Sugar Sensing Using Boronic Acid-Substituted Azobenzenes

In association with increasing diabetes prevalence, it is desirable to develop new glucose sensing systems with low cost, ease of use, high stability and good portability. Boronic acid is one of the potential candidates for a future alternative to enzyme-based glucose sensors. Boronic acid derivatives have been widely used for the sugar recognition motif, because boronic acids bind adjacent diols to form cyclic boronate esters. In order to develop colorimetric sugar sensors, boronic acid-conjugated azobenzenes have been synthesized. There are several types of boronic acid azobenzenes, and their characteristics tend to rely on the substitute position of the boronic acid moiety. For example, o-substitution of boronic acid to the azo group gives the advantage of a significant color change upon sugar addition. Nitrogen-15 Nuclear Magnetic Resonance (NMR) studies clearly show a signaling mechanism based on the formation and cleavage of the B–N dative bond between boronic acid and azo moieties in the dye. Some boronic acid-substituted azobenzenes were attached to a polymer or utilized for supramolecular chemistry to produce glucose-selective binding, in which two boronic acid moieties cooperatively bind one glucose molecule. In addition, boronic acid-substituted azobenzenes have been applied not only for glucose monitoring, but also for the sensing of glycated hemoglobin and dopamine.

Development of a membrane impregnated with a poly(dimethylsiloxane)/poly(ethylene glycol) copolymer for a high-throughput screening of the permeability of drugs, cosmetics, and other chemicals across the human skin.

We aimed to develop a high-throughput screening (HTS) system for preliminary predictions of human skin permeability by using an artificial membrane that can mimic the permeation behaviour of lipophilic and hydrophilic compounds across the human skin. In this study, we synthesized a copolymer containing poly(dimethylsiloxane) (PDMS) and poly(ethylene glycol) (PEG) 6000 and impregnated it onto a supportive membrane filter to prepare a PDMS/PEG 6000 copolymer-impregnated membrane. In addition, we synthesized another polymer without PEG units and used it to prepare an impregnated membrane for determining the role of PEG 6000 units in the PDMS/PEG 6000 copolymer-impregnated membrane. The permeation characteristics of the impregnated membranes were evaluated on the basis of the permeability coefficients of 12 model compounds with different lipophilicities, by using a 2-chamber diffusion cell, and these permeability coefficients were compared with those across the human skin. We obtained a good correlation between the permeability coefficients across the PDMS/PEG 6000 copolymer-impregnated membrane and human skin. Further, we evaluated the permeation characteristics of a 96-well plate model of the PDMS/PEG 6000 copolymer by using 6 model compounds. We obtained an ideal correlation between the permeability coefficients across the PDMS/PEG 6000 copolymer using a 96-well plate and those across the human skin. Thus, the PDMS/PEG 6000 copolymer would be a good candidate for preliminary evaluation of the permeability of lipophilic and hydrophilic compounds across the human skin.

A red fluorophore comprising a borinate-containing xanthene analogue as a polyol sensor

A xanthene derivative containing a borinate moiety emitted red fluorescence with a high quantum yield. The interaction between the borinate and a sugar molecule induced a fluorescence change based on the change in the HOMO-LUMO gap. The response was pH-resistant in a wide range. In addition, catechol quenched through photoinduced electron transfer. The red fluorescence and polyol binding ability of dyes will pave the way for new biological applications of chemical sensors.

Sugar-Responsive Pseudopolyrotaxane Composed of Phenylboronic Acid-Modified Polyethylene Glycol and γ-Cyclodextrin

We have designed a sugar-responsive pseudopolyrotaxane (PPRX) by combining phenylboronic acid-modified polyethylene glycol (PBA–PEG) and γ-cyclodextrin. Phenylboronic acid (PBA) was used as a sugar-recognition motif in the PPRX because PBA reacts with a diol portion of the sugar molecule and forms a cyclic ester. When D-fructose or D-glucose was added to a suspension of PPRX, PPRX disintegrated, depending on the concentration of the sugars. Interestingly, catechol does not show a response although catechol has a high affinity for PBA. We analyzed the response mechanism of PPRX by considering equilibria.

Sugar-responsive pseudopolyrotaxanes and their application in sugar-induced release of PEGylated insulin

We have designed a pseudopolyrotaxane (PPRX), known as a molecular necklace, consisting of phenylboronic acid-modified γ-cyclodextrin (PBA-γ-CyD) and naphthalene-modified polyethylene glycol (Naph-PEG) for developing sugar-responsive insulin delivery systems. Interestingly, structural analyses show that the Naph-PEG/PBA-γ-CyD PPRX obtained by our method was single stranded, whereas ordinary PPRXs using parent γ-CyD were double stranded. The Naph-PEG/PBA-γ-CyD PPRX was poorly water soluble at pH 7.4; however, sugar addition induced disintegration of the PPRX, and the components were dissolved, suggesting that the PBA moiety acts as a sugar sensor. We also have developed a PPRX consisting of Naph-PEG-appended insulin (Naph-PEG-Ins) and PBA-γ-CyD and have confirmed that the release rate of Naph-PEG-Ins was accelerated following sugar addition.Graphical abstract

Sugar-Responsive Layer-by-Layer Film Composed of Phenylboronic Acid-Appended Insulin and Poly(vinyl alcohol)

Previous studies have shown that reversible chemical bond formation between phenylboronic acid (PBA) and 1,3-diol can be utilized as the driving force for the preparation of layer-by-layer (LbL) films. The LbL films composed of a PBA-appended polymer and poly(vinyl alcohol) (PVA) disintegrated in the presence of sugar. This type of LbL films has been recognized as a promising approach for sugar-responsive drug release systems, but an issue preventing the practical application of LbL films is combining them with insulin. In this report, we have proposed a solution for this issue by using PBA-appended insulin as a component of the LbL film. We prepared two kinds of PBA-appended insulin derivatives and confirmed that they retained their hypoglycemic activity. The LbL films composed of PBA-appended insulin and PVA were successfully prepared through reversible chemical bond formation between the boronic acid moiety and the 1,3-diol of PVA. The LbL film disintegrated upon treatment with sugars. Based on the results presented herein, we discuss the suitability of the PBA moiety with respect to hypoglycemic activity, binding ability, and selectivity for D-glucose.

Effect of Physiological Changes in the Skin on Systemic Absorption of Tacrolimus Following Topical Application in Rats

Tacrolimus (TL) ointment is a topical treatment for atopic dermatitis, a disease that exhibits various skin conditions. The effect of skin pathologies on the systemic absorption of TL and related side effects remains unknown. This study aimed to investigate factors affecting the cutaneous absorption of TL. We prepared various skin models in hairless rats by tape stripping, injection of prophlogistic material solution (PMS), and continuous subcutaneous adrenaline (Adr) infusion. In vivo absorption studies were conducted, with measurements of transepidermal water loss (TEWL) and skin blood flow as physiological parameters. Very little TL absorption was observed through intact skin. Greater TL absorption was noted in skins with high TEWL values and fully stripped skin with PMS injections. In contrast, Adr infusion, which reduced skin blood flow, resulted in decreased TL absorption through fully stripped skin. Combined use of TL and Adr on skin with PMS injections resulted in suppression of TL absorption. Our results revealed that TL absorption following topical application is affected by alterations in the skin barrier, blood flow, and vascular permeability. We propose an administration plan for TL in a flowchart as a means of preventing systemic side effects.

Polyol-responsive pseudopolyrotaxanes based on phenylboronic acid-modified polyethylene glycol and cyclodextrins

Phenylboronic acid (PBA), which reacts with polyols to form cyclic esters, was attached to the amino terminal of polyethylene glycol (PEG) via amide bonds. PBA-PEG was used to prepare pseudopolyrotaxanes (pPRXs) by combining it with cyclodextrins (CyDs). In the case of α-CyD, a single stranded pPRX formed that disintegrated in the presence of catechol (CA), d-fructose (Fru), and d-glucose (Glc). The order of response was CA > Fru > Glc, which corresponds with the affinities between the PBA moiety and the polyols. In contrast, a pPRX using γ-CyD, which has a double-stranded structure, showed sugar-induced disintegration but did not show a response to CA. We explained these apparently curious responses of the pPRXs using a mechanism based on the penetrability of the polyol-bound PBA toward the cavities of the CyDs. The pPRXs, which are a class of molecular machine, show two selectivities; one is derived from polyol selectivity, and the other is based on the penetrability for CyDs.

Single-step preparation of topological gels using vinyl-modified β-cyclodextrin as a figure-of-six cross-linker

Herein, we have proposed a single-step preparation of topological gels using vinyl-modified β-cyclodextrin (V-β-CyD) and isoprene. Copolymerization of V-β-CyD and isoprene in an aqueous solution resulted in gelation due to V-β-CyD acting as a novel type of copolymer chain cross-linker. The vinyl moiety of V-β-CyD becomes a part of the copolymer, while the β-CyD moiety of V-β-CyD simultaneously incorporates the isoprene component of the copolymer. V-β-CyD is capable of two different modes of cross-linking at each end, i.e., chemically bonding and mechanically interlocking. Due to the shape of the cross-linking point, we refer to it as figure-of-six cross-linking. Nuclear magnetic resonance analysis showed that the gel contained V-β-CyD and isoprene in an approximately 1:0.3 stoichiometry. The relatively high content of β-CyD was reflected in the character of the gel; the gel swelled in dimethylformamide which is a good solvent of β-CyD. A fluorometric analysis using 6-(p-toluidino)-2-naphthalenesulfonic acid showed that the appended β-CyD was able to accommodate guest molecules. Introduction of an additional vinyl monomer into the gel was also successful. Addition of 4-vinylphenylboronic acid to the preparation procedure yielded a sugar-responsive gel that swelled in the presence of d-fructose.

Related Topic: Measurement of Diffusion Coefficient of Chemicals

Some physical penetration-enhancing techniques may create permeation routes in the skin barrier, which may involve water channels for the hydrophilic drugs. In those processes, diffusion in the channels is the major mechanism of transport, and thus, the diffusion coefficient (D) in aqueous medium could be a primary factor for assessing permeation through the skin barrier. Several experimental methods to determine D for chemical compounds in water or other solvents are described in this chapter.