• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.12-24
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• 1990

Two trypsin-like enzymes, designated trypsin A and 3, purified from the intestine of menhaden by $(NH_4)_2SO_4$ fractionation, Benzamidine-Sepharose 6B affinity chromatography, DEAE-Sephacel ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The two trypsins were subjected to compare the enzymatic properties of the trypsin-like enzymes from the other dark fleshed fishes. Both trypsins catalysed the hydrolysis of N$\alpha$-benzoyl-DL-arginine-p-nitroanilide and they were remarkably inhibited by several well known trypsin-inhibitors, tosyllysyl chloromethyl ketone, soybean trypsin inhibitor, be-nzamidine, leupeptin and antipain, etc. Therefore, it was ascertained that the two enzymes are serine-type trypsins. The molecular weights of these enzymes were about 25,000 and 26,200, respectively, ;Is determined by SDS-PAG electrophoresis and by Sephadex G-100 gel filtration, and the molecular weights of these two enzymes are somewhat fewer than those from the other dark fleshed fishes. Both enzymes had less basic amino acids such as arginine and Iysine, whereas they had slightly high contents of neutral amino acids, glycine, alanine and tryptophane. The enzymes showed a pH optimum of $8\~11$ at $60^{\circ}C$ against the $N\alpha$-benzoyl-DL-argi-nine-p-nitroanilide substrate and they were quite unstable above $40^{\circ}C$ and under the atidic pH region. The Km constant of the two enzymes against the $N\alpha$-benzoyl-DL-arginine-p-nitroanilide was $1.4\times10^{-4}M$ for trypsin A and $4.3\times10^{-5}M$ for trypsin B, respectively.

• Journal of Pharmaceutical Investigation
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• v.22 no.2
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• pp.139-148
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• 1992

A prodrug of cefazolin pivaloyloxymethyl ester (CFZ-PV) was synthesized to improve oral absorption and bioavailability of parent drug by esterification of sodium cefazolin (CFZ) with chloromethyl pivalate. The successful synthesis of CFZ-PV was confirmed by spectroscopic analysis. Partition coefficient studies showed that CFZ-PV is more lipophilic than CFZ. The pharmacokinetic characteristics of CFZ-PV and CFZ preparations were compared following oral administrations of these compounds to rabbits. The analysis of CFZ in plasma was conducted by HPLC method. The ester compound (prod rug) was not detected in plasma following oral administration of CFZ-PV, and although CFZ-PV had not microbiological activity in vitro, the plasma taken after CFZ-PV administration had microbiological activity. From above observations, it was noted that CFZ-PV is rapidly hydrolyzed to CFZ in the body. And it was found that the oral absorption of CFZ-PV was increased, yielding 2-fold higher bioavailability than CFZ. From the results of this experiment, it was concluded that CFZ-PV could be a novel prodrug of CFZ which can improve the oral bioavailability of CFZ.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2004.05a
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• pp.36-37
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• 2004

$\alpha$-Chloromethylnaphthalene is a valuable compound for obtaining of the plant growing stimulator - $\alpha$-napthylacetic acid. Chloromethylation of naphthalene by paraformaldehyde in the presence of glacial acetic acid, phosphoric and hydrochloric acids at temperature 80-85$^{\circ}C$ and duration - 6 hours the $\alpha$-chloromethyl-naphthalene yield was 55-57%. Using Box-Wilson method for mathematical planning of experiment carried out optimization of its synthesis for purpose increasing $\alpha$-chloromethylnaphthalene yield. Preliminary, one - factor experiments were carried out for selecting independence main parameters influencing on the synthesis. A full factor experiment of 2$^3$with extended matrix of planning was used for optimization. Aiming to increase the $\alpha$-chloromethylnaphthalene yield, the obtained mathematical model was used for program of sharp raising on the reply surface. The received optimal conditions for the $\alpha$-chloromethylnaphthalene synthesis were selected as following: molar ratio of naphthalene - parapfsormaldehyde of 1 : 2; temperature -105$^{\circ}C$; duration of the reaction -3 hours. The yield of $\alpha$-chloromethylnaphthalene under these optimal conditions was 75 %.

• Bulletin of the Korean Chemical Society
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• v.28 no.10
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• pp.1725-1728
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• 2007

Mast cells are key effector cells in the early phase allergic inflammation and in diverse immunological and pathological processes. In order to understand the effect on reduction of the anti-dinitrophenyl (DNP) IgE antibody-induced β-hexosaminidase release in RBL-2H3 rat mast cells, a novel series of 4-senecioyloxymethyl- 6,7-dimethoxycoumarins (SMDC) was prepared by reacting 4-chloromethyl-6,7-dimethoxycoumarin with various carboxylic acids. Compounds 8-11 with cyclic moiety such as phenyl, thiophenyl, pyridinyl, and furanyl group were found to inhibit-hexosaminidase release more potently (5.98-9.62 μM) than compounds 3- 7 and 12 with acyclic moiety (19.32-76.78 μM). Furthermore, compounds 8 and 9 inhibited IgE-induced ear swelling and significantly reduced systemic passive cutaneous anaphylaxis reaction in mice.

• BMB Reports
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• v.32 no.6
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• pp.579-584
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• 1999

Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

• Journal of Pharmaceutical Investigation
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• v.25 no.4
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• pp.323-330
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• 1995

Ceftezole pivaloyloxymethyl ester(CFZ-PV) was synthesized to improve oral absorption and bioavailability of parent drug by esterification of ceftezole(CFZ) with chloromethyl pivalate. The successful synthesis of CFZ-PV was conformed by spectroscopic analysis. Partition coefficient studies showed that CFZ-PV is more lipophillic than CFZ. The pharmacokinetic characteristics of CFZ-PV and CFZ were compared following oral administrations of these compounds to rabbits. The amount of CFZ in plasma was determined with an HPLC method. The ester compound (prodrug) was not detected in plasma following oral administration of CFZ-PV, and although CFZ-PV had not microbiological activity in vitro, the plasma taken after oral administration of CFZ-PV had microbiological activity. From above observations, it was noted that CFZ-PV is rapidly hydrolyzed to CFZ in the body. And the oral absorption of CFZ-PV was increased by yielding 2-fold bioavailability rather than CFZ. Therefore, CFZ-PV could be a novel prodrug of CFZ which can improve the oral bioavailability of CFZ.

• YAKHAK HOEJI
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• v.46 no.5
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• pp.313-319
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• 2002

Synthesis of 7$\beta$-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[[(3S, 5S)-5-[4-phenyl-5-(4-methylphenyl or 2-thiophenyl)-4H-l, 2, 4- triazol-3-yl]thiomethylpyrrolidin-3-yl]]thiomethyl-3-cephem-4-carboxylic acids (7a, 7b) were described. (2S, 4S)-4-acethylthio-2-[4-phenyl-5-(4-methylphenyl or 2-thiophenyl)-4 H-1, 2, 4-triazol-3-yl]thiomethyl-1-tert-butoxycarbonylpyrrolidines (4a, 4b) were prepared from trans-4-hydroxy-L-proline with (2S, 4R)-absolute configuration as starting material. 4-Phenyl-5-(4-methylphenyl or 2-thiophenyl)-4 H-l, 2, 4-triazol-3-thiols (2a, 2b) were prepared from p-toluic anhydride and 2-thiophene carboxylic acid hydrazide, respectively. p-Methoxybenzyl 7$\beta$-(Z)-2-(2-for-mamidothiazol-4-yl)-2-(1-tert-butoxycarbonylisopropylimino]acetamido-3-[[ (3S, 5S)-5-[4-phenyl-5-(4-methylphenyl or 2-thio phenyl)-4H-1, 2, 3-triazol-3-yl]thiomethyl-1- tert-butoxycarbonylpyrrolidin-3-yl]]thiomethyl-3-cephem-4-carboxylates (6a, 6b) were achieved by using p-methoxybenzyl ]7P-(Z)-2-(2-formamidothiazol-4-yl)-2-(tert-butoxycarbonylisopropylimino] acetamido-3-chloromethyl-3-cephem-4-carboxylate (5) and (2S, 4S)-4-acethylthio-2-[4-phenyl-5-(4-methyl phenyl or 2-thiophenyl)-4H-1, 2, 4-triazol-3-yl]thiomethyl-1-tert-butoxycarbonyl pyrrolidines (4a, 4b). Removal of formyl, Boc, and p-methoxybenzyl protecting groups were carried out by triflu oroacetic acid and anisole to give the target compounds.

• Journal of environmental and Sanitary engineering
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• v.14 no.1
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• pp.31-41
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• 1999

Several new ion exchange resins have been synthesized from chloromethyl styrene-1, 4-di-vinylbenzene with 1%, 2%, 5% and 10%-crosslinking and macrocyclic ligands of cryptand type by interpolymerization method. The adsorption characteristics and the pH, time, solvents and concentration dependence of the adsorption of metal ions by this resin were studied. The correlation between the separation characteristics of uranium and transition metal on the resins and the stability constants of complexes with macrocyclic ligands have been examined. The resins were very stable in both acidic and basic media and had good resistance to heat at $280^{\circ}C$. The $UO_2^{2+}$ aqueous solution was not adsorbed on the resins below pH 3.0, but the power of adsorption of $UO_2^{2+}$ increased rapidly above pH 4.0. The optimum equilibrium time for adsorption of metallic ions was twenty minutes and adsorptive power decreased in proportion to crosslinking size of the resins and order of dielectric constants of solvents used and the selective sequence for metal cations is in the order of $UO_2^{2+}$, $Cu^{2+}$ and $Ce^{3+}$ .

• Journal of environmental and Sanitary engineering
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• v.15 no.1
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• pp.77-87
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• 2000

Several new ion exchange resins have been synthesized from chloromethyl styrene-l,4-divinylbenzine with 1%, 2%, 10% and 20%-crosslink and macrocyclic ligands of cryptand type by interpolymerization method. The adsorption characteristics and the pH, time, solvents and concentration dependence of the adsorption of metal ions by this resin were studied. The correlation between the separation characteristics of uranium and transition metal on the resins and the stability constants of complexes with macrocyclic ligands have been examined. The resins were very stable in both acidic and basic media and have good resistance to heat. The $UO_2^{2+}$ was not adsorbed on the resins below pH 3.0, but the power of adsorption of $UO_2^{2+}$ increased rapidly above pH 4.0. The optimum equilibrium time for adsorption of metallic ions was two hours and adsorptive power decreased in proportion to crosslink size of the resins and order of dielectric constants of solvents used and the selective sequence for metal cations was in the order of $UO_2^{2+}$, $Ni{2+}$ and $Nd{3+}$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.21 no.2
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• pp.85-96
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• 1988

Purification and some properties of alkaline proteinases in the pyloric caeca of skipjack, Katsuwonus vagans, were investigated. Four alkaline proteinases, temporarily designated proteinases I, II, III and IV, were identified from the tissue extract of the pyloric caeca by ammonium sulfate fractionation, DEAE-Sephadex A-50 chromatography, and Sephadex G-100 and G-200 gel filtration. Result of disc-polyacrylamide gel electrophoretic analysis showed that the purified proteinases II and III were homogenous with the yields of $1.5\%\;and\;1.2\%$, and those specific activities were increased to 33 to 37 fold over that of the crude enzyme solution, respectively. Molecular weight of the proteinases II and III determined by sephadex G-100 gel filtration were 28,500 and 24,200, respectively. The optimum conditions for the caseinolytic activity of the two enzymes were pH 9.6 and $48^{\circ}C$. The reaction rates of the two alkaline proteinases were constant to the reaction time to 80 min in the reaction mixture of $3.4{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The Km values against casein substrate determined by the method of Lineweaver-Burk were $0.56\%$ for proteinase II and $0.30\%$ for proteinase II. The proteinases II and III were inactivated under the presence of $Ag^+,\;Hg^{2+},\;Ni{2+},\;Fe^{2+},\;and\;Cu^{2+}$, and but activated by $Mn^{2+}\;and\;Ca^{2+}$ and markedly inhibited by the soybean trypsin inhibitor and N-p-toluenesulfonyl-L-lysine chloromethyl ketone. Therefore, the proteinases II and III were found to be a group of serine proteases and assured to be trypsin-like proteinases.