Phosphodiesterase inhibitors. Part 6: Design,synthesis, and structure–activity relationships of PDE4-inhibitory pyrazolo[1,5-a]pyridines with anti-inflammatory activity

We previously identified KCA-1490 [(?)-6-(7-methoxy-2-trifluoromethyl-pyrazolo[1,5-a]pyridin-4-yl)-5-methyl-4,5-dihydro-3-(2H)-pyridazinone], a dual PDE3/4 inhibitor. In the present study, we found highly potent selective PDE4 inhibitors derived from the structure of KCA-1490. Among them, N-(3,5-dichloropyridin-4-yl)-7-methoxy-2-(trifluoromethyl)pyrazolo[1,5-a]pyridine-4-carboxamide (2a) had good anti-inflammatory effects in an animal model.     

Separation and Purification of Molecular Species of Galactolipids by High Performance Liquid Chromatography

The NaCl concentration of the growth medium affected hydrogen production by Lyngbya sp. (No. 108) strain. Cells grown in medium containing 3% NaCl produced the most hydrogen. The carbohydrate content of this strain also increased with increasing NaCl concentration of the growth medium up to 720 fig/mg cells at 5 % NaCl. In the presence of 20 finlol/ml MFA (monofluoroacetic acid), inhibition of hydrogen production was observed. We extracted the glycogen from this nonheterocystous filamentous cyanobacterium, Lyngbya sp. (No. 108), and observed that glycogen and carbohydrate consumption of this strain is coincident with hydrogen production.

These results led us to the conclusion that the reserve glycogen or other carbohydrate were used as sources of electron donors for hydrogen production, and that the NaCl concentration of the medium affected the hydrogen production by this strain.     

Correlation Analysis between Eating Quality,Rheological Property and Amylose Content of Starch

A screening was designed to isolate microorganisms having poly(γ-glutamic acid) (PGA) endohydrolase activity. Of the strains screened, TM-4222, from a soil sample, showed the highest viscosity decrement ability on PGA. It was identified to be a Myrothecium sp. The fungal production of the enzyme was slightly promoted with yeast extract and greatly promoted with· both yeast extract and PGA. The fungus was evaluated to produce PGA hydrolase of an endo-type specificity by analyzing of the reaction products.     

Distribution of ω-Amino Acid: Pyruvate Transaminase and Aminobutyrate: α-Ketoglutarate Transaminase in Microorganisms

The distribution of ω-amino acid transaminases in microorganisms was investigated, ω-Amino acid: pyruvate transaminase (ω-APT) was found in bacteria and yeasts, but not in actinomycetes and fungi. On the contrary, aminobutyrate: α-ketoglutarate transaminase (GABA-T) was shown in most of the microorganisms from bacteria to fungi. β-Alanine is a preferred amino donor for the co-APT reaction. Although bacterial and yeast GABA-T are inactive for β-alanine, fungal and actinomycete enzymes react with this compound and γ-aminobutyrate. In comparing these results with those of plant and mammalian enzymes, two different pathways of co-amino acid metabolism are suggested for bacteria, yeast and plants, i.e. one for β-alanine and the other for γ-aminobutyrate, catalyzed by ω-APT and GABA-T, respectively. In actinomycetes, fungi, and mammals GABA-T may be involved in the metabolism of both ω-amino acids. In addition, evolutionary changes of ω-amino acid transaminases are discussed.     

Monohydroperoxides Formed by Autoxidation and Photosensitized Oxidation of Methyl Eicosapentaenoate

Methyl eicosapentaenoate (methyl 5,8,11,14,17-eicosapentaenoate) was subjected to autoxidation and methylene blue sensitized photooxidation. Methyl eicosapentaenoate monohydroperoxides, the primary products of the autoxidation and photosensitized oxidation, were isolated by silica gel column chromatography, and characterized by ultraviolet, infrared and nuclear magnetic resonance spectra. The isomeric composition of the monohydroperoxides were determined by gas chromatography-mass spectrometry as follows: the 5-, 8-, 9-, 11-, 12-, 14-, 15- and 18-isomers (autoxidation), and the 5-, 6-, 8-, 9-, 11-, 12-, 14-, 15-, 17- and 18-isomers (photosensitized oxidation). Methyl eicosapentaenoate was readily oxidized both by autoxidation and by photosensitized oxidation.     

Isolation of Permethylated Dicarbonyl Sugar Derivatives from Polysaccharides

Oxidation of methyl trimethyl glucopyranosides which were obtained by methanolysis of permethylated cellulose, laminarin, and dextran, was performed with dimethyl sulfoxide (DMSO)-phosphorus pentoxide to afford the corresponding ulose derivatives, methyl 2,3,6-tri-O-methyl-d-xylo-hexopyranosid-4-ulose, methyl 2,4,6-tri-O-methyl-d-ribo-hexopyranosid-3-ulose, and methyl 2,3,4-tri-O-methyl-d-gluco-hexodialdo-l,5-pyranoside, respectively, in good or moderate yields. As a new type of derivatives for the linkage analysis of polysaccharides the chromatographic and spectrometric properties of 2,4-dinitrophenylhydrazone of the ulose derivatives were investigated.     

Utilization of n-Paraffins and Vitamin B6 Production by Pichia guilliermondii Wickerham

The accumulation of vitamin B₆ by Pichia guilliermondii Wickerham NK–2 strain grown on hydrocarbon was investigated. Ammonium acetate was more effective than other nitrogen sources tested. Satisfactory utilization by the yeast strain was observed in n-alkanes of C₁₀–C₁₈, and n-pentadecane was the best for vitamin B₆ production. Vitamin B₆ was excreted in the cultural broth mainly in the form of pyridoxal, The maximal vitamin B₆ production was approximately 25 mg per liter of the culture broth.     

Purification and Some Properties of α-Galactosidase from Tubers of Stachys affinis

An α-galactosidase from tubers of S. affinis was purified about 130 fold by ammonium sulfate fractionation, chromatography on DEAE-cellulose and gel filtration on Sephadex G-75. The purified enzyme showed a single protein band on disc gel electrophoresis. The molecular weight of the enzyme was determined to be approximately 42,000 by gel filtration and 44,000 by SDS disc gel electrophoresis. The optimum reaction pH was 5.2. The enzyme hydrolyzed raffinose more rapidly than planteose. The activation energy of raffinose and planteose by the enzyme was estimated to be 7.89 and 11.4 kcal/mol, respectively. The enzyme activity was inhibited by various galactosides and structural analogs of d-galactose. Besides hydrolytic activity, the enzyme also catalyzed the transfer reaction of d-galactosyl residue from raffinose to methanol.