Twenty kinds of derivatives of 2H-pyrido [3, 2-b]-1, 4-oxazin-3-one (8a-12d) having β-dialkylaminoethyl groups were synthesized by the ring closure of 2-amino-3-hydroxypyridine (1) with α-halogeno acid ester in the presence of sodium hydride in DMF. The following results were obtained from the preliminary investigation on the local anesthetic and the analgesic activities (mouse writhing test) of these synthesized compounds. It was proved that the duration time of the local anesthetic activities of almost all these compounds is inferior to that of cocaine or lidocaine, but the duration of these activities, in quite few cases, superior to that of cocaine or lidocaine ; the analgesic action of some of them are apporoximately equivalent to that of pyrabital.
Twenty five kinds of derivatives of 4-(3-dialkylamino-2-hydroxypropyl)-2H-pyrido-[3, 2-b]-1, 4-oxazin-3-one (9a-13e) were synthesized by three different methods, and obtained as hydrochloride. The following results were obtained from the preliminary investigation on the local anesthetic and the analgesic activities (mouse writhing test) of these synthesized compounds. Almost all of the compounds were inferior to cocaine or lidocaine on the local anesthetic action ; these were approximately equivalent to or slightly weaker than pyrabital.
Ethyl 5-hydroxybenzo [α] phenazine-6-carboxylates (IIa, b) were prepared by the condensation of ethyl 1, 4-dihydro-1, 4-dioxo-3-methoxy-2-naphthoate (I) with o-phenylene-diamines. Acylations and alkylations of IIa, b gave the corresponding o-acyl and alkyl compounds (IIIa-k). The reaction of IIa, b with ethanolamine gave β-hydroxyethyl-carboxamides (Va, b). Compound Va was converted into β-chlorinated compound (VIa), whose chlorine was substituted by dialkylamines to give ethylenediamine-type amides (VIIa, b, c). Compounds VIId and VIIe were obtained by the reaction of IIb with dialkylalkylenediamines. Oxidation of IIa, b afforded N-oxides (IXa, b). The antimicrobial activities of all synthetic compounds were tested by paper discagar diffusion assay. Compounds IIId, e, g, VIIa and IXa showed inhibitory activities to T. interdigitale and IXa to C. albicans.
New saponins, named gypenosides I (1), II (2), V (5), VI (6), VII (7), IX (9), X (10), XI (11), XIII (13), XIV (14), besides the known ginsenosides-Rb1 (3), -Rb3 (4), -Rd (8) and -F2 (12), were isolated from Gynostemma pentaphyllum MAKINO. On enzymatic hydrolysis, all saponins yielded 20-O-β-D-glucopyranoside of 20 (S)-protopanaxadiol (compound K) as a prosapogenin. The configuration of the sugar linkages in each saponin was established on the basis of chemical and spectroscopic evidence.
Hog pancreatic kallikrein A and B were separated by DEAE-Sephadex A-50 chromatography and purified by preparative high performance liquid chromatography (TSK GEL G2000 SW column). Kallkrein A and B were homogeneous in SDS-polyacrylamide gel electrophoresis. Antisera were elicited in rabbits by immunization of these purified antigens. The precipitin lines formed during diffusion of kallikrein A and B against the antisera, using double diffusion method, were completely comfluent and relative cross reactivity of respective antisera were quite similar. The Fab'fragments were conjugated with β-D-galactosidase using N, N-o-phenylene-dimaleimide as a coupling reagent. Using this antibody-enzyme complex, it was possible to determine as little as 1 ng/ml of kallikrein by the sandwich enzyme immunoassay. The results obtained with various samples of kallikrein (K-1000, K-200, K-100 and K-25) were in accord with the activities of biological assay.
The inclusion complexes of 7-substituted coumarin derivatives (7-hydroxy-, 7-amino-, 7-dimethylamino-, 7-methoxy-, and 7-ethoxy-4-methylcoumarins) with β-cyclodextrin in aqueous solutions were formed. The equilibrium constants of these complexes were obtained by fluorescence and circular dichroism spectrophotometric methods. Fluorescence enhancement or quenching was observed upon inclusion of coumarin derivatives with β-cyclodextrin. The fluorescent behavior was discussed from the standpoint of the solvent effect of coumarin derivatives and of the geometry of the inclusion complexes assumed from their 1H nuclear magnetic resonance spectra.
A new arginine ester hydrolyzing enzyme in the human urine (human urinary arginine esterase, HUAE) was found. The content levels of this new enzyme, determined by N-α-tosyl-L-arginine methyl ester (Tos-Arg-Me) hydrolyzing activity, were measured to be 0.34 to 0.86 nmol/min/ml of human urine and the percentage of content level of this enzyme was calculated to be about 10 to 20 per cent of the total Tos-Arg-Me hydrolyzing activity. HUAE was partially purified and then separated into two forms (HUAE-1 and -2) by DEAE-cellulose chromatography. The approximate molecular weights of HUAE-1 and -2 were estimated to be 4.8×104 and 3.0×104 by Sephadex G-100 gel filtration, respectively.
Emulsions of a relatively small amount of oleic acid in aqueous glycyrrhizin (GA) solution were investigated by means of turbidity measurement. Oleic acid was easily emulsified in aqueous GA solution by shaking with a reciprocating shaker to give stable o/w emulsions. With increasing GA concentration at a constant pH and ionic strength, the turbidity of emulsion initially increased and then decreased at GA concentrations above its critical micelle concentration. This effect may by attributed to the solubilization of oleic acid by GA solution. The emulsifying power of GA, which was defined as the change of turbidity per unit volume of oleic acid emulsified, was enhanced with rising pH and decreasing ionic strength of GA solution. The dissociation of both GA and oleic acid was considered to play an important role in the emulsification of the system.
Antimicrobial activities of N-arylsulfonylthioureas (23 compounds) and the corresponding isothioureas (28 compounds) were examined against bacteria and fungi parasitic to animals and influenza A2/Adachi virus in vitro. Remarkable activities were not observed in these compounds, but some thioureas showed MIC of 12.5μg/ml against Trichophyton asteroides and Trichophyton interdigitals, and some isothioureas MIC of 25μg/ml against Mycoplasma gallisepticum. N-Phenyl-N'-(p-tolylsulfonyl) thiourea and S-butyl-N-methyl-N'-(p-tolylsulfonyl) isothiourea showed the inhibitory activity against the virus but not the virucidal activity. Fifty % inhibitory concentrations (Index) of these two compounds were 74 (13.5) and 40 (17.7) μg/ml, respectively.
The size and its distribution of the particles reduced by wet grinding (pulverization by intervention) of five kinds of solid drugs using mortar and pestle or a ball mill were determined. The results obtained were compared with those by dry grinding and a number of conclusions were drawn as in the following way : 1. d-Camphor which belongs to a plastic crystal was easily pulverized in the presence of ether ; however, once the solvent was evaporated, masses were formed by continued impaction due to plasticity and the particle size became progressively enlarged. Accordingly, it is certain that the liquid ether among particles prevent plastic conglomeration of d-camphor. 2. Size-reducibility of d-camphor in the presence of potato starch by dry trituration with mortar and pestle was examined. As was expected from the above, the size of d-camphor could effectively reduced because the starch particles acted as a grinding aid by gathering around those of d-camphor. 3. It was demonstrated by low temperature differential scanning calorimetry that the heat of fusion of d-camphor was abnormally small as compared with the heat of transition at -29°C ; accordingly, its fusion entropy was estimated to be 2.9 cal deg-1 mol-1, and was much smaller than those of the ordinary organic crystals. 4. Next to d-camphor, wet grinding of l-menthol with ether or ethanol was found effective. In this case, however, size reduction was rather attributable to the fine crystals separated from the very thick solution of l-menthol in these solvents. Since on continued ball-milling after evaporation of the solvent, it is certain that l-menthol crystals have some plasticity and that the intervented solvent will also take part in reduction of the particle size by preventing conglomeration. 5. It is evident the wet grinding is an unnecessary procedure for the size reduction of brittle crystals such as the ones of ethyl p-aminobenzoate, salicylic acid, and aspirin, since the particles of them were equally reduced either by wet or by dry trituration.
We improved a measurement of gastric emptying in rat, in which the remaining of the gastric contents was estimated as amounts of quinaldine red mixed with the given test meal. The following results were obtained by using this method ; when chemicals were injected subcutaneously, gastric emptying was significantly faoilitated by carbamylcholine and bombesin under both anesthetized and conscious conditions, and delayed by isoproterenol under both conditions. Carbamylcholine-induced facilitation in anesthetized rats was inhibited by atropine, and isoproterenol-induced suppression in conscious rats was inhibited by propranolol. However, histamine and norepinephrine did not affect gastric emptying. Tannic acid or oxethazaine which was orally administered with the test meal, caused a suppression of gastric emptying. Acidification or neutralization of the test meal by HCl or Al (OH)3 did not affect gastric emptying. It seems that gastric emptying was suppressed in the acetic acid gastric ulcer in the rat, but it was facilitated in the duodenal ulcer.
To optimize a solvent system systematically for liquid-solid chromatography separation, the strength indices of stronger solvents such as diethyl ether (B1), ethyl acetate (B2), acetone (B3), tetrahydrofuran (B4), dioxan (B5) and 2-propanol (AB) were determined on the basis of a linear relationship between the logarithm of the capacity ratios and the logarithm of the solvent composition in binary systems containing n-hexane (O) as a diluent. A new procedure for the graphic schematization of the relative strength and equi-eluotropic composition of stronger solvents is elaborated. The experimental results obtained with triterpenoid sapogenins were compared with the Snyder's solvent strength parameter and the Neher's calculation for predicting the equi-eluotropic solvent composition and the differences among three strength parameters are discussed.
In order to identify the constituents of capsicum annuum tincture and related additives in cosmetic preparations, the liquid-solid chromatographic resolution of capsaicine analogues was examined. For this purpose, binary solvents consisting of n-hexane as a diluent and various proton donor and acceptor solvents such as chloroform, diethyl ether, ethyl acetate, dioxan and ethanol as stronger components were used along with silica gel and chemically bonded packing columns. An eluent containing dioxan afforded maximum resolution for the group of solutes. Better results were obtained at lower temperatures. A phase system consisting of n-hexane-dioxan and silica gel at 0°C and incorporating a fluorescence detector was applied following a common clean-up procedure to the analysis of nonanoic acid vanillylamide, an additive in a hair tonic.
The retention indices of 17 drugs and metabolites including hypnotics, antiepileptics, analgesics, and central nervous system stimulants in liquid-liquid chromatography were systematically examined to devise a highly efficient extraction and fractionation procedure in biological fluid analysis. A high performance system incorporating an acid-treated silica gel column and a UV-detector was assembled to determine the capacity ratios in aqueous two phase organic solvents consisting of diethyl ether and n-hexane (1 : 0, 7 : 3, 1 : 1, 3 : 7 v/v). The corrected capacity ratios (k'cor) of the drugs in an aqueous organic phase system were calculated from the experimental retention data and phase ratios in liquid-liquid chromatography. A linear correlation between the logarithm of the k'cor value and the logarithm of solvent composition in a liquid-liquid partition system was demonstrated in this study.
To characterize and identify hetero-aromatic compounds systematically, the liquid chromatographic retention behavior of 19 pyridine and pyrazine derivatives was investigated using various packing materials. A linear correlation between the logarithms of capacity ratio and the logarithms of binary solvent composition was obtained for weaker polar solutes and solvents by silica gel liquid-solid chromatography. Reversed-phase (RP) systems were required for the separation of higher polar solutes such as pyrazine N, N-dioxides. A non linear correlation between retention indices and the methanol-water composition was observed for octadecylsilyl-silica packing. This suggests a complex retention mechanism including the participation of a silanol group on the silica gel surface. On the other hand, an ideal linear correlation between the logarithm of the capacity ratio and solvent composition was obtained in the RP system using polystyrene (PS) gel. The PS gel reversed-phase system seems to be involved in a simpler retention mechanism than the chemically bonded-silica gel phase system and consequently more suited for the determination of basic nitrogen compounds.
In this paper piezoelectric studies are described with the films of polyimine derivatives which are presumed to have a preferential orientation of dipole groups in the molecular chains if they are extended and poled. The film of polyimine derived from isophthalaldehyde and 3, 3'-thiodipropionic acid dihydrazide showed quite high piezoelectric constant 0.67×10-7 cgs esu. The results of dynamic modulus and pyroelectric coefficient measurements are also described.
Mannich reaction of o- or p-hydroxyacetophenone was examined by reacting of o-hydroxyacetophenone (1) or p-hydroxyacetophenone (2) with paraformaldehyde and piperidine under pressure. Mannich reaction of 2 gave 4-hydroxy-β-piperidinopropiophenone (3), 4-hydroxy-3-piperidinomethylacetophenone (4) and 4-hydroxy-3-piperidinomethyl-β-piperidinopropiophenone (5) ; the reaction of 1 gave 3-piperidinomethyl-4-chromanone (6) and 3-hydroxymethyl-3-piperidinomethyl-4-chromanone (7).