Masaaki Ueki

Removal of 9-fluorenylmethyloxycarbonyl (Fmoc) group with tetrabutylammonium fluoride

Abstract Tetrabutylammonium fluoride in N, N-dimethylformamide (DMF) is an effective alternative to the piperidine reagent for the removal of 9-fluorenylmethyloxycarbonyl (Fmoc) group in the solid phase peptide synthesis.

Inhibitory effects of spinorphin, a novel endogenous regulator, on chemotaxis, O−2 generation, and exocytosis by N-formylmethionyl-leucyl-phenylalanine (FMLP)-stimulated neutrophils

To characterize the inflammatory effect of spinorphin, an endogenous peptide purified from bovine spinal cord, its effects on chemotaxis, O2- generation, and exocytosis by N-formylmethionyl-leucyl-phenylalanine (FMLP)-stimulated human neutrophils (PMNs) in vitro were examined. At 10 microM, spinorphin significantly inhibited chemotaxis by FMLP-stimulated PMNs. Spinorphin at 100 microM also inhibited both O2- generation and exocytosis of beta-glucuronidase and collagenase by FMLP-stimulated PMNs. The mechanisms by which spinorphin inhibits these PMN functions were examined further. Spinorphin markedly suppressed the binding of FML[3H]P to its receptor on PMNs, as observed in a binding assay. However, other neuropeptides that were examined (angiotensin II and substance P) had no effect on FML[3H]P binding, suggesting the possibility that spinorphin plays a specific role in the inhibition of the binding between FMLP and its receptor. The suppression of FMLP binding also caused a decrease of the FMLP-induced intracellular calcium concentration [Ca2+]i, which acts as a second messenger leading to PMN functions. These results suggest that spinorphin may be a new endogenous inflammation-regulatory peptide that modulates the interaction of FMLP with its receptor.

Inhibitory action of spinorphin, an endogenous regulator of enkephalin-degrading enzymes, on carrageenan-induced polymorphonuclear neutrophil accumulation in mouse air-pouches

Whether spinorphin, an endogenous regulator of enkephalin-degrading enzymes, plays a role in an anti-inflammatory action was examined, using a mouse air-pouch assay as a model of acute inflammation. Repeated intravenous administration (6 times) of spinorphin every hour significantly suppressed carrageenan-induced polymorphonuclear neutrophil (PMN) accumulation, an indicator of inflammation (3.21+/-0.95 x 10[6] cells vs 8.92+/-0.96 x 10[6] cells, 10mg/kg spinorphin-treated vs saline-treated group, n=5, P<0.01) at 6 hr. The combination of spinorphin and leuhistin (2 mg/kg, i.v.), a specific inhibitor of aminopeptidase N (APN), markedly suppressed the PMN accumulation induced by carrageenan (1.11+/-0.17 x 10[6] cells, 88% inhibition compared to the saline-treated group, n=5, P<0.01). This inhibition was less than, but comparable to that of dexamethasone (30 mg/kg/one shot, i. v.), a representative anti-inflammatory drug. These results indicate that spinorphin may be an endogenous anti-inflammatory regulator, its inhibitory activity being modulated by APN.

Synthesis and anti-HIV activity of nonatyrosine N- and O1-9-decasulfate.

To develop a potent and effective anti-HIV compound with a definite polyanionic structure, synthesis of oligotyrosine sulfates by oligomerization with simultaneous sulfation of tyrosine was tried. One component was successfully isolated from the mixture containing many products as its sodium salt (Y-ART-4) and was identified as the salt of nonatyrosine N- and O1-9-decasulfate, NaO3S-[Tyr(SO3Na)]9-ONa. Anti-HIV activity of Y-ART-4, determined from the protection it provided against HIV-induced cytopathic effects, was almost the same with that of dextran sulfate and curdlan sulfate.

Solid phase synthesis and biological activities of [Arg8]-vasopressin methylenedithioether

Solid phase synthesis of [Arg8]-vasopressin methylenedithioether, an analog of vasopressin which contains an extra methylene group between the two sulfur atoms of Cys1 and Cys6, is described. Methylene insertion occurred easily when the thiol free peptide on a solid support was treated with tetrabutylammonium fluoride in dichloromethane at room temperature for 3 h. The uterotonic in vitro, pressor, and antidiuretic activities of the compound were reduced in comparison to [Arg8]-vasopressin by one order of magnitude.

Aharonov–Bohm-type Effects in Triangular Antidot Lattice

Three kinds of Aharonov–Bohm (AB)-type oscillation have been investigated in triangular antidot lattice fabricated from a GaAs/AlGaAs two-dimensional electron gas sample. The oscillation periods of Altshuler–Aronov–Spivak (AAS) effect and AB-type effect near zero magnetic field are determined by the unit cell area, whereas those of AB-type oscillations in the quantum Hall plateau transition regime are governed by the effective area of antidot. The evolution of the high-field AB-type oscillation as a function of gate voltage gives infomation on the profile of the self-consistent potential associated with compressible edge channels formed around antidot. The temperature dependences and decoherence mechanisms of the AAS and AB-type oscillations near zero magnetic field as well as the high-field AB-type oscillation are discussed.

Selective removal of phenacyl ester group with a TBAF.xH2O-thiol system from amino acid derivatives containing benzyl or 4-nitrobenzyl ester

A facile cleavage method of phenacyl ester bonds by combined use of tetrabutylammonium fluoride hydrate (TRAF·xH2O) and 1-octanethiol was established. Benzyl and 4-nitrobenzyl esters are almost stable toward this reagent.

Sulfated pentagalloyl glucose (Y-ART-3) inhibits HIV replication and cytopathic effects in vitro, and reduces HIV infection in hu-PBL-SCID mice.

To evaluate the efficacy of Y-ART-3 as an antiviral drug for HIV infections, its anti-HIV activity was assessed in vitro in cell culture systems and in vivo in hu-PBL-SCID mice. The results indicated that Y-ART-3 invariably inhibited not only HIV-1, but also HIV-2 and SIV strains. Its mechanism of action is ascribed to inhibition of viral adsorption to CD4-positive cells. In an in vivo study, human Ig- and CD4-positive cells were detected at similar levels in Y-ART-3-treated hu-PBL-SCID mice that were infected with HIV, and in PBS-treated control hu-PBL SCID mice that were not infected with HIV. If HIV positivity was calculated using the number of tests in which HIV was detected (i.e. PCR, and p24 from co-cultures of spleen and peritoneal wash cells), a significant effect of Y-ART-3 at a dose of 4 mg/kg was noted. Therefore, Y-ART-3 may be considered to be a potent and effective anti-HIV compound.

Methylphosphinyl (Dmp): A new protecting group of tyrosine suitable for peptide synthesis by use of boc-amino acids

Abstract Use of dimethylphosphinyl (Dmp) group as a side-chain phenolic OH protecting group of tyrosine in peptide synthesis was studied. The Dmp group is resistant to trifluoroacetic acid and hydrogenolysis and removed by fluoride ion or liquid HF.

Transformation of acyloin O-acyl derivatives to ketones using tetrabutylammonium fluoride-thiol system

Abstract The treatment of acyloin (α-hydroxyketone) O -acyl derivatives with tetrabutylammonium fluoride hydrate (TBAF·xH 2 O)-1-3,propanedithiol system gave the corresponding ketones. Further, it was found that N -methylmorpholine added could prevent the concomitant of unidentified compounds to give the pure products in high yields.