Biomedical subjects

Shiro Kobayashi

Publications and source records attributed to Shiro Kobayashi.

44 records · Page 3Linked to original sources

PubMedDate not supplied

Enzymatic synthesis and antioxidant properties of poly(rutin).

Rutin, quercetin-3-rutinoside, is one of the most famous glycosides of flavonoid and widely present in many plants. In this study, we performed an oxidative polymerization of rutin using Myceliophthora laccase as catalyst in a mixture of methanol and buffer to produce a flavonoid polymer and evaluated antioxidant properties of the resultant polymer. Under selected conditions, the polymer with molecular weight of several thousands was obtained in good yields. The resulting polymer was readily soluble in water, DMF, and DMSO, although rutin monomer showed very low water solubility. UV measurement showed that the polymer had broad transition peaks around 255 and 350 nm in water, which were red-shifted in an alkaline solution. Electron spin resonance (ESR) measurement showed the presence of a radical in the polymer. The polymer showed greatly improved superoxide scavenging activity and inhibition effects on human low-density lipoprotein (LDL) oxidation initiated by 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH), compared with the rutin monomer. The polymer also protected endothelial cells from oxidative injury induced by AAPH as a radical generator with a much greater effect than the rutin monomer.

Animals↗

PubMedDate not supplied

Amplification of antioxidant activity of catechin by polycondensation with acetaldehyde.

Catechin exhibits numerous biological and pharmacological effects attributed to antioxidant action. The synthetic poly(catechin)s condensed through acetaldehyde with different molecular weights were assessed in terms of antioxidant activity and enzyme inhibitory activity on the basis of a catechin repeating unit and compared with monomeric catechin. The poly(catechin)s showed great amplification of superoxide scavenging activity, xanthine oxidase (XO) inhibitory activity, and inhibition effects on human low-density lipoprotein oxidation initiated by 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) as a radical generator on the catechin unit level, compared to monomeric catechin: these activities were proportional to their molecular weights. The reducing power of the polymer was lower than that of monomeric catechin, which decreased with increasing the molecular weight. The polymer also protected endothelial cells from oxidative injury induced by AAPH, with a greater effect expressed on a catechin unit basis than that of the monomer. These results demonstrate that the poly(catechin)s are more potent antioxidant agents and enzyme inhibitors.

Acetaldehyde↗

PubMedDate not supplied

New tyrosinase inhibitors, (+)-catechin-aldehyde polycondensates.

In this study, new tyrosinase inhibitors, (+)-catechin-aldehyde polycondensates, have been developed. Tyrosinase is a copper-containing enzyme that catalyzes the hydroxylation of a monophenol (monophenolase activity) and the oxidation of an o-diphenol (diphenolase activity). In the measurement of tyrosinase inhibition activity, (+)-catechin acted as substrate and cofactor of tyrosinase. On the other hand, the polycondensates inhibited the tyrosine hydroxylation and L-DOPA oxidation by chelation to the active site of tyrosinase. The UV-visible spectrum of a mixture of tyrosinase and the polycondensate exhibited a characteristic shoulder peak ascribed to the chelation of the polycondensate to the active site of tyrosinase. Furthermore, circular dichroism measurement showed a small red shift of the band due to the interaction between tyrosinase and the polycondensate. These data support that the polycondensate acts as an inhibitor of tyrosinase.

Agaricales↗

PubMedDate not supplied

Superoxide anion scavenging and xanthine oxidase inhibition of (+)-catechin-aldehyde polycondensates. Amplification of the antioxidant property of (+)-catechin by polycondensation with aldehydes.

In this study, the antioxidant property of (+)-catechin-aldehyde polycondensates has been examined. Superoxide anions are one of the most typical reactive oxygen species (ROS) and generated by xanthine oxidase (XO). The measurements of the superoxide anion scavenging and XO inhibition activity showed that catechin had pro-oxidant properties in lower concentrations and little XO inhibition. On the other hand, the polycondensates exhibited much higher effects compared to the catechin monomer, and their physiological activities were greatly affected by the structure of polycondensates. Steady-state analysis of the inhibition against XO showed that the inhibition type of the polycondensate was uncompetitive. Furthermore, the results of the circular dichroism and UV-visible measurements of a mixture of the polycondensate and XO were in good agreement with that of the steady-state analysis; the spectral changes due to the chelation of the polycondensate onto the Fe/S and/or the FAD center of XO were observed. These data strongly suggest that the polycondensates possess a great potential as antioxidant for various applications.

Aldehydes↗

PubMedDate not supplied

Polymerization of polyfunctional macromolecules: synthesis of a new class of high molecular weight poly(amino acid)s by oxidative coupling of phenol-containing precursor polymers.

Oxidative coupling of phenol-containing precursor poly(amino acid)s, poly(alpha-glutamine), poly(alpha/beta-asparagine), and poly(gamma-glutamine) derivatives, has been examined to produce a new class of soluble poly(amino acid)s. Under appropriate reaction conditions, the Fe-salen and HRP catalysts efficiently induced the oxidative coupling without formation of insoluble gels, yielding the soluble polymers of high molecular weight. The oxidative coupling behaviors were greatly influenced by the structure and phenol content of the precursor polymer. The selection of the substrate concentration and catalyst amount was crucial for the production of soluble polymers of high molecular weight.

Amino Acids↗

PubMedDate not supplied

Bottom-up synthesis of hyaluronan and its derivatives via enzymatic polymerization: direct incorporation of an amido functional group.

This paper reports the synthesis of hyaluronan (HA) and its derivatives via the hyaluronidase-catalyzed polymerization of 2-substituted oxazoline derivative monomers designed as "transition-state analogue substrates". Polymerization of 2-methyl oxazoline monomer from N-acetylhyalobiuronate (GlcAbeta(1-->3)GlcNAc) effectively proceeded at pH 7.5 and 30 degrees C, giving rise to synthetic HA (natural type) in an optimal yield of 78% via ring-opening polyaddition under total control of regioselectivity and stereochemistry. Hyaluronidase catalysis enabled the polymerization of 2-ethyl, 2-n-propyl, and 2-vinyl monomers, affording the corresponding HA derivatives (unnatural type) with N-propionyl, N-butyryl, and N-acryloyl functional groups, respectively, at the C2 position of all glucosamine units in good yields. The 2-isopropyl oxazoline derivative provided the N-isobutyryl derivative of HA in low yields. Monomers of 2-phenyl and 2-isopropenyl oxazoline derivatives were not polymerized. The mechanism of the polymerization is discussed.

Amides↗

PubMedDate not supplied

Amplification of inhibitory activity of catechin against disease-related enzymes by conjugation on poly(epsilon-lysine).

A new inhibitor against disease-related enzymes, collagenase, hyaluronidase, and xanthine oxidase, has been developed by the laccase-catalyzed conjugation of catechin on poly(epsilon-lysine). The resulting poly(epsilon-lysine)-catechin conjugate showed greatly improved inhibition effects on activity of these enzymes, whereas the catechin monomer showed very low inhibition activity. The kinetic analysis on the inhibition of collagenase exhibited that the conjugate was a mixed-type inhibitor. The amplified activities might offer high potential as a therapeutic agent for prevention of various enzyme-related diseases.

Catechin↗

PubMedDate not supplied

Enzymatic synthesis of chondroitin 4-sulfate with well-defined structure.

Synthesis of chondroitin sulfate (ChS) with well-defined structure was achieved for the first time by hyaluronidase-catalyzed polymerization. N-Acetylchondrosine (GlcAbeta(1-->3)GalNAc) oxazoline derivatives sulfated at C4 (1a), C6 (1b), and both C4 and C6 (1c) in the GalNAc unit were synthesized as transition state analogue substrate monomers for hyaluronidase (HAase) catalysis. Compound 1a was effectively polymerized by the enzyme, giving rise to synthetic ChS sulfated perfectly at the C4 position in all N-acetylgalactosamine units (Ch4S, 2a) in good yields. Molecular weights (Mn) of 2a ranged from 4000 to 18,400, which were controlled by varying reaction conditions. Compounds 1b and 1c were not catalyzed by the enzyme, affording the corresponding disaccharides through the oxazoline ring-opening without formation of polysaccharides.

Animals↗