Pyrrothines, known as broad-spectrum antibiotics, showed potent membrane stabilization and platelet anti-aggregant activities. Thiolutin, the most potent among them, is three times as active as indomethacin in HH, APA and CPA, but has no activity in the PD system. Pyrrothines exhibited strong inhibitory activity on 5-HT release from bovine platelets, which may be responsible in part for their platelet anti-aggregant activity.

抄録全体を表示

Dialkyl 4-aryl-1, 4-dihydro-2, 6-dimethylpyridine-3, 5-dicarboxylates (III) were subjected to the Mannich reaction with excess paraformaldehyde and secondary amine hydrochloride in boiling ethanolic solution. When the compounds III were treated with a 5-to 7-fold molar excess of the aldehyde and amine salt, 2, 6-bis (2-disubstituted aminoethyl)-dihydropyridine derivatives (V) were obtained in good yields. On treatment with a 2- to 3-fold molar excess of the reactants, compounds III were converted to 2-(2-disubstituted aminoethyl)-6-methyldihydropyridine derivatives (IV) and V. When dioxane was used as a solvent with a 6-fold molar excess of the reactants, the tetrakis (dimethylaminomethyl) derivative (VII) was obtained.

抄録全体を表示

A method for the synthesis of 2-amino-4-phenyl-3, 4-dihydrothieno [2, 3-d] pyrimidine derivatives (12 and 13b) is described. Reduction of the 3-substituted 2, 3-dihydrothieno- [2, 3-d] pyrimidin-2-one (4c) and -2-thione (9b, c) with sodium borohydride gave the 1, 2, 3, 4-tetrahydro derivatives (6c and 10b, c), whereas the 3-unsubstituted compound 4a could be reduced only with lithium aluminum hydride to afford the 1, 2, 3, 4-tetrahydro derivative 6a. The 2-chloro derivative (7) was reduced with sodium borohydride to give the 2-chloro-3, 4-dihydro derivative 8a, which was reacted with ethyl bromoacetate to yield predominantly the 3-substituted derivative (8c). Amination of 8c gave compound 12. Methylation of 10b and 10c, followed by amination similarly yielded 13b and 12, respectively. In this amination some oxidative products (14) were also obtained. Compounds 12 and 13b had lower inhibitory effects against blood platelet aggregation than the quinazoline analogs (1) and (2).

抄録全体を表示

A stable nitroxide radical containing active methylenes, 2, 2, 6, 6-tetramethyl-4-piperidone-1-oxyl (I), was found to be carboxylated by potassium phenoxide and carbon dioxide in an aprotic solvent (N, N-dimethylformamide), producing new nitroxide compounds. The mono- or dicarboxylate was formed selectively, depending on the molar ratio of the substrate to potassium phenoxide. The ultraviolet and visible spectra of the monomethyl- (II) and dimethylester (III) of the carboxylates as well as those of I and 2, 2, 6, 6-tetramethylpiperidine-1-oxyl (IV) were investigated in various solvents. The results indicated that II exists mainly as the keto form in a polar solvent, and as the enol form in a nonpolar solvent, while the enol form of III is predominant in all the solvents tested. The nitrogen hyperfine coupling constants, A_N, of I-IV in the electron spin resonance spectra were determined, and the results are discussed in terms of the keto-enol equilibrium. Furthermore, the distribution coefficients of these nitroxide radicals between hexane and water were measured to assess the potential suitability of these newly formed nitroxide radicals for use as spin labels or probes.

抄録全体を表示

Condensation of 1, 6 : 2, 3-dianhydro-β-D-mannopyranose with 2-methyl-(3, 4, 6-tri-O-acetyl-1, 2-dideoxy-α-D-glucopyrano)-[2', 1' : 4, 5]-2-oxazoline (2) in the presence of a catalytic amount of anhydrous p-toluenesulfonic acid in boiling 1, 2-dichloroethane afforded crystalline 4-O-(2-acetamido-3, 4, 6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl)-1, 6 : 2, 3-dianhydro-β-D-mannopyranose (3) in 42% yield. Azidolysis of the oxirane ring of 3, reduction of the azido to an amino group, and N-acetylation afforded 2-acetamido-4-O-(2-acetamido-3, 4, 6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl)-3-O-acetyl-1, 6-anhydro-2-deoxy-β-D-glucopyranose (6) in 50% yield. Compound 6 was indistinguishable from the product prepared in 52.2% yield by direct condensation of 2-acetamido-3-O-acetyl-1, 6-anhydro-2-deoxy-β-D-glucopyranose with 2. Acetolysis of 6 provided the title disaccharide.

抄録全体を表示

Co (III)-bovine carbonic anhydrase was reduced by L-ascorbic acid with recovery of the enzyme activity. The rate determined by following the increase in the absorbance at 640 nm (characteristic absorption of Co (II)-bovine carbonic anhydrase) was completely consistent with that determined from the recovery of the esterase activity. This result indicates that the recovery of the enzyme activity with L-ascorbic acid was caused by the reduction of a cobalt ion in the enzyme from Co (III)-bovine carbonic anhydrase to Co (II)-bovine carbonic anhydrase by L-ascorbic acid. The rate of the reduction of Co (III)-bovine carbonic anhydrase has a first-order dependence on the concentrations of both L-ascorbic acid and Co (III)-bovine carbonic anhydrase, and the rate constant is 3.6-7.2×10^-1 sec^-1M^-1.

抄録全体を表示

A synthesis of the title trisaccharide (14) is reported. The first step of the synthetic route is stereospecific condensation of 2-acetamido-1, 6-anhydro-3-O-benzyl-2-deoxy-β-D-glucopyranose with 2-methyl-(3, 4, 6-tri-O-acetyl-1, 2-dideoxy-α-D-glucopyrano)-[2', 1' : 4, 5]-2-oxazoline to yield a fully protected β-D-(1→4)-linked disaccharide (8) bearing 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc) at the non-reducing residue. After acetolysis of the 1, 6-anhydro ring of 8, the resulting fully-protected disaccharide is debenzylated to yield chitobiose heptaacetate (10) having a free hydroxyl group at the C-3 position of the reducing GlcNAc. Compound 10 is glycosylated with 2, 3, 4-tri-O-benzyl-α-L-fucopyranosyl bromide by a bromide ion-catalyzed reaction. After removing the protecting groups of the resulting trisaccharide by debenzylation and de-O-acetylation, an anomeric mixture of 14 is obtained as an amorphous solid. CMR spectral data for 14 are also presented.

抄録全体を表示

In order to examine the feasibility of size reduction by heat transition of polymorphic drugs, crystals of the α-form of sulfathiazole were heated above its transition temperature and the physico-chemical and micromeritical properties of the product were studied by IR spectroscopy, X-ray diffractometry, DSC, BET gas adsorption analysis, and electron microscopy. The original α-crystals became opaque after transition and the polymorphic form of the product was identified as the β-form of sulfathiazole. Corresponding to the loss of transparency, many cracks and fissures were observed electron-microscopically on the surface. At the same time, a considerable increase of the surface area was found. The convex sections between the cracks and fissures can be ascribed to plastic deformation induced by the stress of transition. In spite of the remarkable changes in the surface appearance, the β-particles were unexpectedly stable on standing. The extents of the lattice disorder measured by X-ray diffractometry were found to be small. When ball-milling was applied to the β-crystals obtained by heat-induced transition, the particle size was further reduced. However, they became less stable and retransition was induced more easily at room temperature. In addition, lattice disorder attributable to the mechano-chemical effect of grinding was observed in the ground α-and β-powder.

抄録全体を表示

Six symmetrical linear octiphenyls were synthesized by the Ullmann homo-coupling reaction of iodoquaterphenyl. Another satisfactory method for synthesizing octiphenyls is also described, i.e., a Kharash-type Grignard cross-coupling of biphenylylmagnesium bromide and diiodoquaterphenyl in the presence of bis (acetylacetonato) nickel (II). Moreover, m-septiphenyl was successfully synthesized by deamination of the corresponding amino compound. Infrared studies of the polyphenyls indicated that the fine-structure bands in the regions of 770-810 and 870-915 cm^-1 may be considered as indicators of the presence of consecutive m-phenylene rings, regardless of the existence of o-or p-phenylene rings. The ultraviolet spectra of the octiphenyls, which contain two kinds of linkages, showed absorption curves closely related to those of quaterphenyls corresponding to the structural units of the former compounds. The nuclear magnetic resonance spectra of the polyphenyls without an o-phenylene ring showed multiplet peaks due to the resonances of an isolated m-phenylene proton at the lowest field within a narrow region (δ7.81-7.95). Huckel molecular orbital calculations of the longest wavelength absorption bands of ten polyphenyls were carried out. The calculated and observed wavelengths were in rather good agreement except for the cases of three compounds.

抄録全体を表示

Grignard reaction of 2, 3, 4-tri-O-benzyl-D-xylopyranose yielded a single product with very high stereoselectivity. A threo relationship of the newly created chiral center with respect to C₂ was established by converting the product into the δ-lactone of D-ido configuration, then to the unsaturated lactone of threo configuration. The result made possible a stereoselective synthesis of 6-deoxy-D-idose from D-xylose in a small number of steps. Models accounting for the high stereoselectivity in the Grignard reaction of aldoses are discussed.

抄録全体を表示

Treatment of 4-acetoxy-2-azetidinone (9) with various sodium alkyl trithiocarbonates gave 4-[(alkylthio) thiocarbonyl] thioazetidinones 10, which were converted into the phosphoranes 8. Intramolecular Wittig reactions of 8 followed by deblocking yielded penem-3-carboxylic acids 5.

抄録全体を表示

Treatment of 14-bromo-6-dehydroxysantoninic acid (1a) with 10% Na₂CO₃-acetone solution gave the lactone dimer (2), the spiro dimer (5), and a hydroxy compound (4), in 17%, 40%, and 21% yields, respectively. The molecular structure of the major product, 6-dehydroxysantoninic acid spiro dimer (5), was confirmed by X-ray crystallographic analysis. The mechanism of formation of 5 during the course of this reaction is discussed.

抄録全体を表示

A mucous polysaccharide, named Narcissus-T-glucomannan, was isolated from the bulbs of Narcissus tazetta L. var. chinensis ROEMER. The final preparation was homogeneous as determined by ultracentrifugal analysis, glass-fiber electrophoresis, and gel chromatography. It was composed of D-mannose and D-glucose in the molar ratio of 5 : 1, and its molecular weight was estimated to be 119000. The O-acetyl groups in the glucomannan was identified and the content amounted to 22.7%. They were located at positions 6 and 2, 6 of most of the D-mannose units. Methylation, periodate oxidation, partial acetolysis, and enzymatic degradation studies showed that the glucomannan is mainly composed of β-1→4 linked aldohexopyranose residues, and that it contains about 42 aldohexose units per non-reducing group on average. D-Mannose units occupy non-reducing terminal positions and branching points linked through position 3.

抄録全体を表示

The tert-butyldimethylsilyl group was found to be effective for protection of the hydroxyl group in penem synthesis. Manipulation of the side-chain hydroxyl group of base-sensitive penem-3-carboxylates was successfully conducted by means of a displacement reaction using a combination of triphenylphosphine-diethyl azodicarboxylate in the presence of a nucleophile.

抄録全体を表示

Total syntheses of (±)-phomenone (5a), (±)-3-epiphomenone (12), (±)-ligularenolide (27), and (±)-furanoligularanone (34) are described. Dehydration of 6 followed by epoxidation gave the epoxide (8), which was treated with lithium diethylamide to afford the alcohol (9). Epoxidation of 9 gave the α-epoxide (10), which was deketalized to give the diketone (11) Reduction of 11 with NaBH₄ gave (±)-3-epiphomenone (12) as a major product. Stereoselective synthesis of (±)-5a starting from 7 was examined. Deketalization of 7 gave the triene-dione (13) which was reduced with NaBH₄ to afford the alcohols 14a and 14b. Epoxidation of 3α-ol (14a) gave the epoxide (15), which was treated with lithium diethylamide to afford the diol (16). Epoxidation of 16 with hydrogen peroxide gave (±)-phomenone (5a) regio- and stereoselectively. Treatment of 17 with lithium diisopropylamide followed by condensation with methyl pyruvate gave the hydroxy ester (19), which was treated with acetic acid to give the ketone (24). 24 was thioketalized to give 25, which was treated with p-toluenesulfonic acid to afford the unsaturated lactone (26). 26 was transformed to (±)-ligularenolide (27) and (±)-tetrahydroligularenolide (28) in good yield. Condensation of 17 with acetol pyranyl ether followed by catalytic hydrogenation gave 33. Treatment of 33 with p-toluenesulfonic acid gave (±)-furanoligularanone (34), which was transformed to (±)-3β-furanoligularanol (36).

抄録全体を表示

From Lycopodium wightianum collected in Borneo, two new triterpenoids named wightianol-A and wightianol-B were isolated together with α-onocerin, tohogenol, and lycoclavanin. The structures of the new compounds were established as 14β-serratane-3β, 14β, 20β, 21β, 24-pentol (4a) and serrat-14-ene-3β, 20β, 21β, 24-tetrol (5a), respectively. For comparison, the C₃-epimer of lycoclavanin, 16-oxoserrat-14-ene-3β, 20β, 21β, 24-tetrol (14), was prepared and characterized as its tetraacetate.

抄録全体を表示

In an attempt to clarify the effects of drugs on the metabolism, transport mechanism and life time of human erythrocytes, the effects of chlorpromazine and clemastine on the ATP content, sodium pump and glucose-6-phosphate (G-6-P) dehydrogenase were studied. Chlorpromazine at 2 and 3×10^-4M caused only a 5% reduction in the ATP level of erythrocytes and the drug at 8×10^-4 and 10^-3M induced 20 and 22% decreases in the amount of the ghost membrane, respectively. The incubation of erythrocytes in an ATP-generating system had only a slight protective effect against the drug-induced hemolysis. The Na⁺, K⁺-ATPase activity in the ghost membrane was markedly decreased by chlorpromazine at 6×10^-4 to 10^-3M without inhibition of Mg²⁺-ATPase activity, and inhibition of the Na⁺, K⁺-ATPase activity was observed a short time after the drug treatment. There was also a partial inhibition of G-6-P dehydrogenase activity by the drugs. Complete inhibition of Na⁺, K⁺-ATPase activity with ouabain resulted in no increment of the directly drug-induced hemolysis and no extreme shape changes. Therefore, the directly drug-induced hemolysis is not necessarily an energy-dependent process. However, the cells exposed to chlorpromazine at prelytic concentrations were gradually hemolyzed upon prolonged incubation in isotonic solution at 37°. Consequently, the drug treatment appears to shorten the survival time of the cells, probably due to inhibition of cellular metabolism and to membrane damage.

抄録全体を表示

A competitive enzyme immunoassay for phenobarbital with alkaline phosphatase as a label is described. p-Aminophenobarbital was used to prepare the immunogen and the alkaline phosphatase conjugate. The IgG fraction prepared from anti-phenobarbital antiserum was bound to Sepharose 4B or adsorbed on polystyrene tubes. The solid phase antibody could not discriminate p-hydroxyphenobarbital from phenobarbital, but had almost no cross-reactivity with metharbital, barbituric acid, thiobarbiturates and other antiepileptic drugs. The ranges of phenobarbital concentration measurable by enzyme immunoassay with Sepharose-IgG (EIA-I) and with polystyrene tube-IgG (EIA-II) were 10 to 100 ng/tube and 100 to 1000ng/tube, respectively. The results obtained by radioimmunoassay correlated well with those obtained by EIA-I (γ=0.995) and EIA-II (γ=0.943). Intra-and inter-assay variations of EIA-II were determined by replicate assays of sera containing 20 μg/ml and 80 μg/ml phenobarbital. The intra-assay coefficients of variation were 3.6% and 5.8%. The inter-assay coefficients of variation were 11.2% and 9.2%. Because of the convenience of the EIA-II procedure, the method is suitable for the routine assay of serum phenobarbital in small clinical laboratories.

抄録全体を表示

O-Acylation of primary aliphatic nitro compounds with acid chlorides in N, N-dimethylacetamide led to the formation of nitrile oxides, which cyclized in situ with olefinic dipolarophiles in a 1, 3-dipolar cycloaddition mode to afford 2-isoxazoline derivatives. Optimum reaction conditions were investigated, and the use of several types of dipolarophiles, such as acetylenes, Schiff bases, aromatic nitriles, and ketones, in this cycloaddition resulted in the formation of the corresponding cycloadducts, namely isoxazoles, 1, 2, 4-oxadiazolines, 1, 2, 4-oxadiazoles, and 1, 4, 2-dioxazoles, respectively, in reasonable yields.

抄録全体を表示

Dissolution profiles of indomethacin/polyvinylpyrrolidone (PVP) coprecipitates (involving a crystallization process) were investigated kinetically by the dispersed amount and rotating disk methods in comparison with those of physical mixtures. The dissolution rate constant, k_t, the rate constant of the crystallization process, k_r, and saturated concentrations before and after the crystallization, C_SM and C_SO, were calculated from the dissolution data by a curve fitting procedure. The effects of the content and molecular weight of PVP in the coprecipitates on these dissolution parameters were also investigated in detail. It was found that increase of the content and the molecular weight of PVP correlated well with retardation of the crystallization process. In order to obtain desirable dissolution patterns, we attempted to estimate the optimum content of PVP in the coprecipitates by analysis of the dissolution parameters.

抄録全体を表示

N-(2-Hexahydropyrimidinoethyl) anilines (6a-f) were prepared by the condensation of N-(β-bromoethyl) aniline hydrobromide and hexahydropyrimidines. They were further converted to N-(2-hexahydropyrimidinoethyl) propionanilides by N-propionylation. The analgesic activities of the twelve compounds thus obtained were examined by subcutaneous administration to mice. Among the propionanilides, the N-isopropyl- and N-benzylhexahydropyrimidine derivatives (7c, e) possessed ca. 1/3 and 1/6 of the analgesic effect of pentazocine, respectively, and 7e showed a lower toxicity. On the other hand, the N-phenethyl derivative (7f) was inactive.

抄録全体を表示

N-(1-Methyl-2-piperazinoethyl) anilines (5a-e) and 2-alkoxy-6-[1-methyl-2-(4-phenethylpiperazino) ethyl]-amino-benzothiazoles (8a-d) were prepared by the reduction of 1-(2-anilinopropionyl) piperazines (4a-e) and 1-[2-(2-alkoxy-benzothiazolyl-(6))-aminopropionyl]-4-phenethylpiperazines (7a-d). N-(1-Methyl-2-piperazinoethyl) propionanilides (6a-e) and 2-alkoxy-6-[N-[1-methyl-2-(4-phenethylpiperazino) ethyl] propionamide]-benzothiazoles (9a-d) were prepared by N-propionylation of 5a-e and 8a-d. Analgesic activity testing showed that (A) N-[1-methyl-2-(4-benzylpiperazino) ethyl]-propionanilide (6d) and N-[1-methyl-2-(4-phenethylpiperazino) ethyl] propionanilide (6e) possessed ca. 1/3 of the analgesic effect of pentazocine ; (B) N-propionylation of N-[1-methyl-2-(4-benzylpiperazino) ethyl] aniline (5d) and N-[1-methyl-2-(4-phenethylpiperazino) ethyl]-aniline (5e) increased the analgesic activity, but N-propionylation of 8a-d decreased the analgesic activity ; (C) an aniline derivative (6e) was more potent than the 2-alkoxy-6-aminobenzothiazole derivatives (9a-d).

抄録全体を表示

Malondialdehyde (MDA) reacted with secondary amines (dimethylamine, diethylamine, piperidine, pyrrolidine and morpholine) at 37° under mild acidic or neutral conditions to yield β-dialkylaminoacroleins (1-5) of trans, s-trans conformation. The optimal pH of the reaction was 3-5, and the yields were 15-55% in a 6 hr incubation. The acroleins (1-5) were unstable under acidic and alkaline conditions, and produced a pink color on reaction with 2-thiobarbituric acid. β-Dimethylaminoacrolein (1) could be readily nitrosated in the acidic pH range to produce N-nitrosodimethylamine, and the rate of nitrosamine formation from 1 at pH 5.0 was much higher than that from dimethylamine ; this is consistent with earlier observations of the stimulating effect of MDA on nitrosamine formation.

抄録全体を表示

A new peptide that improves the rhythmicity of cultured myocardial cell clusters, designated as AAP (antiarrhythmic peptide), has been discovered and isolated in a pure form from bovine atria by extraction with boiling water at pH 4.5, followed by column chromatography procedures, two-dimensional paper chromatography and electrophoresis. The yield was 200 μg per 1.0 kg of wet tissues. The homogeneity of this peptide was confirmed by the chromatographic and/or electrophoretic behavior of the native peptide and its dansyl derivative. It was found that AAP is a hexapeptide composed of hydroxyproline (1), proline (1), glycine (3) and alanine (1), from the results of amino acid analysis and apparent molecular weight determination on a Sephadex G-15 column. The NH₂-terminal amino acid was glycine as determined by dansylation, and the COOH-terminus was also glycine (carboxypeptidase A digestion and dansylation after hydrazinolysis of AAP). This peptide changed the rhythmic beatings with continuous cellular fibrillation of myocardial cell clusters induced by 0.7 mM potassium, 3 mM calcium or 0.05 mM ouabain in modified Eagle's minimum essential medium supplemented with 2.5% bovine serum to normal rhythmic beatings, and changed irregular beatings with cellular fibrillation at 0.5 mM potassium or 5 mM calcium to rhythmic beatings with cellular fibrillation, followed by normal beatings. This peptide also restored the arrhythmic movements of isolated rat atrium induced by low potassium concentration and addition of acetylcholine to a normal rhythm, in the same way as quinidine. The activities of this peptide on myocardial cells and isolated atrium were stronger than those of quinidine. No antiarrhythmic activity was found in amino acids, peptides and proteins chemically related to AAP, such as hydroxyproline, proline, glycine, alanine, Gly-Pro, Gly-Pro-Leu, Gly-Pro-Leu-Gly-Pro, gelatin and heat-treated gelatin.

抄録全体を表示

In order to elucidate the physiological significance of bovine ventricle protein (BVP) and antiarrhythmic peptide (AAP) obtained from bovine heart, we investigated their effects on spreading, beating, arrhythmic movements, macromolecular synthesis, continuous cultivation and calcium incorporation of cultured rat myocardial cells in Eagle's minimum essential medium (MEM) supplemented with or without serum. Both BVP and AAP at 10^-7M stimulated the spreading phenomenon and the protein synthesis of myocardial cells in serum-free culture, and promoted prolonged survival with spontaneous beating in culture with 1% serum. As regards the beating properties of myocardial cells, they had different effects. AAP, which improved the arrhythmic movements induced by low potaasium, high calcium or addition of ouabain, depressed the beating rate and the ratio of beating cells at 10^-7-10^-6M. However, BVP promoted beating even in continuous cultivation over 10 days, but did not affect the arrhythmic movements induced by low potassium or high calcium at 10^-7-10^-6M. When myocardial cells were cultured in modified Eagle's MEM-0.5% bovine serum albumin at 0.5 mM potassium, 5 mM calcium or 0.2 mM ouabain (which induced arrhythmic movements of cultured cells), their spreading was significantly suppressed. Quinidine, oxytocin and insulin, which improve arrhythmia in the intact heart, stimulated the spreading phenomena and improved the arrhythmic movements induced by low potassium as well as AAP. AAP also decreased the incorporation of ⁴⁵Ca into myocardial cells, but BVP did not have this effect. It is concluded that AAP shows antiarrhythmic action as a result of depressions of calcium incorporation and potassium effusion, with low excitation and prolonged survival of myocardial cells. BVP shows prolonged survival with spontaneous and strong beating in continuous culture for 10 days.

抄録全体を表示

Synthesis of porcine motilin, which exhibits a gastric motor stimulating activity, was accomplished by the solid-phase method with diphenyl phosphorazidate (DPPA) and diethyl phosphorocyanidate (DEPC) as coupling reagents. The purified synthetic peptide showed characteristic contractile activity towards the duodenum, colon and jejunum in the rabbit. Its potency was almost the same as that of the natural material.

抄録全体を表示

Two new dibenzocyclooctadiene lignans, tigloylgomisin P (1), and angeloylgomisin P (2) were isolated from the fruits of Schizandra chinensis BAILL. (Schizandraceae). Their absolute structures were elucidated by means of chemical and spectral studies.

抄録全体を表示

The conversion of a hydroxymethyl group at the 2- or 4-position of simple pyrimidines to a chloromethyl, cyanomethyl, ethoxycarbonylmethyl, or formyl group is described. Various pyrimidines having an olefinic side chain were also synthesized via the Wittig reagents derived from chloromethylpyrimidines.

抄録全体を表示

In order to obtain specific antisera for use in radioimmunoassay and enzyme immunoassay of cortisol, new hapten-carrier conjugates were prepared from 4-hemisuccinoyloxycortisol, 4-(carboxymethylthio) cortisol, 4-(2-carboxyethylthio) cortisol and 4-(2-hemisuccinoyloxyethylthio) cortisol by coupling with bovine serum albumin (BSA) employing the N-hydroxysuccinimide ester method or mixed anhydride method. The specificity of anti-cortisol antisera elicited in rabbits by immunization with these antigens was tested by cross-reaction studies with closely related steroids and by measuring the amount of cortisol in urine specimens by means of radioimmunoassay. Comparison of the specificity of these antisera with that of other antisera prepared with BSA conjugates of cortisol 3-(O-carboxymethyl) oxime and 21-hemisuccinoylcortisol was also carried out. The results showed that the antisera obtained were sufficiently specific and thus that the C-4 position was suitable for the attachment of a carrier protein for use in the production of antibodies.

抄録全体を表示

A new method for the determination of serum uric acid was developed. In this method, uric acid is converted to allantoin, hydrogen peroxide and carbon dioxide by uricase (EC 1.7.3.3) and then allantoin is measured colorimetrically at 525 nm after condensation with diacetylmonoxime-thiosemicarbazide in 6N hydrochloric acid. In order to apply this method to serum, the coexisting urea is removed by urease (EC 3.5.1.5) treatment before the color reaction. Application of the method to patients'sera showed a good correlation with the results of a standard uricase-UV method ; the regression equation was y=0.954x+0.290 (mg/100 ml) and the correlation coefficient was γ=0.930. A characteristic feature of the new method is that it is not influenced by such commonly encountered interfering substances as ascorbic acid (160 mg/100 ml), glutathione (100 mg/100 ml), glucose (500 mg/100 ml) and bilirubin (10 mg/100 ml).

抄録全体を表示

A sensitive and accurate UV-assay procedure based on multiple linear regression analysis (MLR method) was developed for the measurement of uric acid with uricase. The absorbance at 290 nm of uric acid as a function of time was calculated from the color points of a mixture of uric acid, an intermediate (1-carboxy-2, 4, 6, 8-tetraazabicyclo [3.3.0] octa-4-ene-3, 7-dione) and the final product (allantoin) by the simplified complementary tristimulus colorimetry (SCTS method). The MLR method allowed the evaluation of both initial and final absorbances at the same wavelength with high accuracy. The average recovery of uric acid added to an aqueous solution was 99.9±0.02%, and the coefficient of variation was less than 3% (n=4) even at low concentration. The assay could also be performed directly on serum samples without deproteinization. An average recovery of 103.1±0.7% was obtained for 0.17-1.33 mg/dl of uric acid in serum solutions, and the coefficients of variation of 0.32 mg/dl and 0.41 mg/dl were 1.6% (n=4) and 2.4% (n=4), respectively.

抄録全体を表示

A sensitive fluorimetric method for the assay of dopamine β-hydroxylase in rat plasma is described. Octopamine, formed enzymatically from the substrate tyramine, is separated by chromatography on a Dowex 50W-X4 column and oxidized with periodate to p-hydroxybenzaldehyde, which is then quantitated by means of the fluorimetric method for selective determination of aromatic aldehydes with 2, 2'-dithiobis (1-aminonaphthalene). The method is readily performed with good precision and is suitable for the assay of many samples simultaneously.

抄録全体を表示

The 3-, 7- and 12-monosulfates of bile acids having an oxo or acetoxyl group on the steroid nucleus have been synthesized. Cholic acid derivatives appropriately protected were sulfated with chlorosulfonic acid and pyridine in the usual manner. The utilization of partial acetylation and hydrolysis together with chromium trioxide oxidation and sodium borohydride reduction provided the desired sulfates of cholic acid derivatives in satisfactory yields. The nuclear magnetic resonance spectral properties of the bile acid derivatives are briefly discussed.

抄録全体を表示

Sulfonation of the benzene ring of 3-(p-toluidino)-1-oxo-2-propene-2-sulfonic acid (III) (6.11×10^-5M) in 99.4% sulfuric acid at 25° was observed by following the UV spectral change of the solution. In a 9.51×10^-1M solution in 99.4% sulfuric acid, however, the bulk of III remained unchanged, and on dilution with 93.8% sulfuric acid, 6-methyl-quinoline (II) was formed. Sodium 4-(3-oxo-1-propenylamino) benzenesulfonate (V), sodium 4-(3-oxo-1-propenylamino)-3-methylbenzenesulfonate (VI) and sodium 2-(3-oxo-1-propenylamino)-5-methyl-benzenesulfonate (VII) were synthesized in connection with this investigation.

抄録全体を表示

The reversible sulfonation at the α-position of β-(p-chloroanilino) acrolein (V) in sulfuric acid at various concentrations was studied kinetically. The rate constants of cyclodehydration of β-(p-toluidino) acrolein (I) with reversible sulfonation at the α-position were evaluated by non-linear least-squares analysis using the rate constants of the reversible sulfonation of V as approximate initial values for those of I. The rate constant k₁ of cyclodehydration of I is related to Hammett's acidity function H₀ as expressed by the equation log k₁+H₀=-13.97. The rate of cyclodehydration of I is one-fortieth of that of 4-(p-toluidino)-3-penten-2-one (VIII) in sulfuric acid at 25°.

抄録全体を表示

The decomposition products of hydrazones of hydralazine with pyruvic acid, acetone and α-ketoglutaric acid were studied. The pyruvic acid hydrazone and acetone or α-ketoglutaric acid hydrazone were converted to 3-methyl-s-triazolo [3, 4-a] phthalazine (MTP) and phthalazinone, respectively, both of which are known hydralazine metabolites. New high pressure liquid chromatographic (HPLC) methods for determining hydralazine and MTP were developed. The formation of MTP and the disappearance of hydralazine at a physiological concentration ratio and pH at 37° were studied by the HPLC methods. The formation of MTP followed second-order kinetics, and its rate constant was determined.

抄録全体を表示

The thymosin α₁ C-terminal fragment H-Lys-Asp-Leu-Lys-Glu-Lys-Lys-Glu-Val-Val-Glu-Glu-Ala-Glu-Asn-OH (positions 14-28) was synthesized by a conventional method. Two thymosin α₁ fragments, H-Lys-Lys-Glu-Val-Val-Glu-Glu-Ala-Glu-Asn-OH (positions 19-28) and the pentadecapeptide fragment synthesized in this study, were tested for effect on lymphocyte transformation in the uremic state. The synthetic pentadecapeptide increased ³H-thymidine incorporation into DNA in the uremic state, but the synthetic decapeptide had no effect on the ³H-thymidine incorporation.

抄録全体を表示

The chemical introduction of 4-thiouridine (S⁴U) residues into nucleic acids was carried out by the reaction of nucleic acids with liquid hydrogen sulfide in aqueous pyridine, which caused the conversion of cytosine moieties to 4-thiouracil moieties.

抄録全体を表示

The interactions of Sepharose-bound bovine neurophysin I and II with oxytocin and arginine-vasopressin were studied. The amounts of hormones bound to immobilized neurophysin II were found to depend strongly on temperature, being larger at 4° than at 36°. It was found that oxytocin and vasopressin were eluted in different fractions from a column of Sepharose-neurophysin I by shallow pH gradients in either direction from pH 5.8, whereas the hormones emerged in the same fractions from a Sepharose-neurophysin II column.

抄録全体を表示

Rat pancreatic elastase was purified on a column of lima bean trypsin inhibitor-Sepharose after inactivating trypsin and chymotrypsin in the pancreatic homogenate by treatment with N^α-p-tosyl-L-lysylchloromethyl ketone and N-tosyl-L-phenylalanylchloromethyl ketone. The molecular weight of the enzyme was 25000, as determined by polyacrylamide gel electrophoresis. A radioimmunoassay method was established for rat elastase ; the enzyme level could be determined over the range of 2.5-300 ng of the enzyme per ml.

抄録全体を表示

The adsorption of benzothiadiazines by carbon black from aqueous solution was investigated in detail. The adsorption isotherms obtained were well described by the Langmuir equation. It was shown that the adsorbability was positively related to the relative diuretic activity. The adsorption was found to be reversible with change of temperature in a perturbation experiment, suggesting a physical adsorption mechanism. The pH of the buffer solution had no clear effect on the adsorption of benzothiadiazines near the neutral pH region. It was shown qualitatively by thin-layer chromatography that the hydrolysis of benzothiadiazines was accelerated by carbon black.

抄録全体を表示

The photochemical and thermal reactions of 3-diazothiochroman-4-one (2a), 3-diazochroman-4-one (2b), 3-diazo-1-methyl-1, 2, 3, 4-tetrahydroquinolin-4-one (2c), and 2-diazo-1-tetralone (2d) were investigated. Wolff rearrangement was observed with 2a, 2c, and 2d only under photolytic conditions. In the case of 2a, migration of the thioether group competed with the above reaction.

抄録全体を表示

The bulbs of Lycoris radiata HERB. (Amaryllidaceae) were found to contain a new phenolic base, O-demethyllycoramine (O-demethyldihydrogalanthamine) (3), as well as previously isolated alkaloids, such as pretazettine (1), lycorine (4), lycoramine (5), lycorenine (6), demethylhomolycorine (7), hippeastrine (8), and homolycorine (9). The structure of 3 was confirmed by demethylation of lycoramine (5) with pyridine hydrochloride.

抄録全体を表示

Aniline hydroxylation with a hemin-cysteine system was studied under various conditions as a model for cytochrome P-450 enzymes. The reaction was characterized by using various inhibitors, such as hydroxyl radical scavengers, a singlet oxygen trapping agent, superoxide dismutase and catalase. The system was compared with simple model systems consisting of cupric ion, ascorbic acid and hydrogen peroxide or the Udenfriend system. The hemin-cysteine model system was not inhibited appreciably by any of the inhibitors tested. On the basis of these findings and the fact that the thiolate-heme iron linkage was found to be present in the hemin-cysteine system, a possible mechanism for hydroxylation by the model system is proposed.

抄録全体を表示

The two isomers of 5-(2-amino-1-hydroxybutyl)-8-hydroxycarbostyril (2), which is a major metabolite of procaterol, were synthesized.

抄録全体を表示

8-Hydroxycarbostyril (4b), which is a starting material for the synthesis of procaterol, was synthesized by two new routes.

抄録全体を表示

9-Allyladenine 1-oxide (3) has been prepared from 1-ethoxyadenine (6) in 58% overall yield through an unequivocal synthetic route. The route consists of an initial allylation of 6 with allyl bromide and Et-O bond cleavage of the resulting 9-allyl-1-ethoxyadenine hydrobromide (7) by treatment with boiling pyridine. Replacement of allyl bromide by 3-methyl-2-butenyl bromide in the above reaction sequence afforded 9-(3-methyl-2-butenyl) adenine 1-oxide (9) in 59% overall yield through the 1-ethoxy derivative 8.

抄録全体を表示

A potent mutagen, ethyl N-ethyl-N-nitrosocarbamate, was isolated from the reaction of ethyl carbamate with NaNO₂. A possible reaction mechanism was proposed.

抄録全体を表示

Several modified derivatives of glycyrrhetinic acid were prepared for obtaining compounds which are devoid of aldosterone-like properties and retain or enhance the therapeutic activities of the mother compound. Among them, olean-12-en-3β, 30-diol showed antiallergic and antiulcerous activities without inhibition of Δ⁴-5α- and Δ⁴-5β-reductases of 3-keto-Δ⁴-steroids. This compound was prepared in a good yield from glycyrrhetinic acid by reduction with sodium bis (2-methoxyethoxy) aluminum hydride followed by catalytic hydrogenation of the intermediate with Pd-C.

抄録全体を表示

Two new diterpenes were isolated from the rhizomes of Alpinia speciosa K. SCHUM. (Zingiberaceae) and their structures were established by the spectral evidences as I and II. The latter has an unusual bisnorlabdane carbon skeleton. It is the first example that diterpenes were obtained from Alpinia genus.

抄録全体を表示