To clarify the mode of action of erythromycin on eucaryotic cells, the morphological and biochemical changes of Cochliobolus miyabeanus with erythromycin were studied and following results were obtained. (1) The potato-sucrose medium containing 2000 μg/ml of erythromycin showed 50% inhibition of this fungal growth. (2) The mycelium grown in the presence of erythromycin (2000 μg/ml) was fixed in 2% KMnO4, dehydrated by the usual method, and subjected to scanning electron microscopic observation (Hitachi SSM-II). Most of these mycelia showed morphological changes and looked like the beans of a rosary, whereas thiamphenicol and cycloheximide, which are inhibitors of protein synthesis like erythromycin, did not cause the same morphological changes. (3) Analysis of mycelia of the fractionated into soluble carbohydrate, nucleic acid, lipid, and chitin-like subsatnce showed very little differences between the mycelia grown in the presence and absence of erythromycin. (4) The enzyme activities concerned in the synthesis or decomposition of cell-wall main components (β-glucan and chitin-like substance) were not affected by the addition of erythromycin while a little inhibition of proteolytic enzyme activities was seen by the presence of erythromycin.
Reactions of 4, 5-diethoxy-2-methyl (and phenyl)-3 (2H)-pyridazinone (I) with sodium ethoxide, 10% aqueous sodium hydroxide, and 47% hydrobromic acid were investigated. In some cases, the preferential displacement of 4-ethoxy group by hydroxyl group was observed, indicating that the displacement on the 3 (2H)-pyridazinone ring is affected by the nature of substituent at position 2, reaction species, and the medium enployed.
The flow properties of mixed powder of two components were investigated. Three kinds of magnesium aluminosilicate (MAS), which have different particle sizes, were preparated for use as an additive, and potato starch was used as a base particle. The fluidity was estimated by the measurement of the angle of repose. It was found that the optimal fluidity was obtained at some proportion of the additive, and this optimum proportion varies with particle size and moisture content of MAS. For example, optimum proportion was 0.75% by weight for MAS-I (7.72 μm in mean particle size) and 0.18% by weight for MAS-III (1.63 μm in mean particle size), A significant decrease was observed in optimum proportion for heat-treated MAS (200-400°, 1 hr) and steam-treated MAS (92% R.H.). The reason for the former seems to be adhesiveness due to activity of the performed dehydroxyl sites, and rehydration occurring on the surface of the base particle. In the latter, it seems that adsorbed water molecules on the surface of additives build bridges between MAS and base particle, and the contact between base particles is prevented by the additive particles.
Since there is little report on the photo-oxidation of tocopherols, known as the natural antioxidant, photo-oxidation of a model compound of tocopherols was carried out and its products were examined. Reaction of 6-hydroxy-2, 2, 5, 7, 8-pentamethylchroman (1) in benzene afforded 6.8% of trimer (2), 9.5% of 5-formyl-6-hydroxy-2, 2, 7, 8-tetramethylchroman (3), and 5.6% of 2-(3-hydroxyisoamyl)-3, 5, 6-trimethyl-p-benzoquinone (4), while that in ethanol solution gave 6.8% of 2, 2.5% of 5-ethoxymethyl-6-hydroxy-2, 2, 7, 8-tetramethylchroman (6), and 25.7% of 4. The reaction of 6-hydroxy-2, 2, 5, 8-tetramethylchroman (7) in benzene gave 9.0% of 6-hydroxy-2, 2, 5, 8-tetramethyl-7-(2, 2, 5, 8-tetramethyl-6-chromanyloxy) chroman (8), 8.9% of trimer (9), 13.5% of 5-formyl-6-hydroxy-2, 2, 8-trimethylchroman (10), and 6.4% of 3, 6-dimethyl-2-(3-hydroxyisoamyl)-p-benzoquinone (11). Similarly, 6-hydroxy-2, 2, 7, 8-tetramethylchroman (12) gave 6.8% of 6-hydroxy-2, 2, 7, 8-tetramethyl-5-(2, 2, 7, 8-tetramethyl-6-chromanyloxy) chroman (13) and 26.6% of 6-hydroxy-2, 2, 7, 8-tetramethyl-5-(6-hydroxy-2, 2, 7, 8-tetramethylchroman-5-yl) chroman, while 6-hydroxy-2, 2, 8-trimethylchroman (16) gave 6.2% of 6-hydroxy-2, 2, 8-trimethyl-5-(2, 2, 8-trimethyl-6-chromanyloxy) chroman (17) and 8.5% of 6-hydroxy-2, 2, 8-trimethyl-5-(6-hydroxy-2, 2, 8-trimethylchroman-5-yl) chroman (18) The methyl group in 5-position of 1 and 7 seems to be active and 2, 3, 9, and 10 formed by the reaction progressing with the benzyl radical so formed were obtained. In 7, 12, and 16, in which the postion ortho to 6-hydroxy group is vacant, compounds 8, 13, 14, 17, and 18 formed by the carbon-oxygen coupling or carbon-carbon coupling were obtained.
Oxidation of the model compounds of tocopherols with bis (salicylidene) ethylenediiminocobalt (1) was examined. Oxidation of 6-hydroxy-2, 2, 5, 7, 8-pentamethylchroman (2) in methanol gave 12.8% of trimer (3), 8.3% of spirodienonedimer (4), 3.7% of 1-(6-hydroxy-2, 2, 7, 8-tetramethylchroman-5-yl)-2-[5, 6-dimethyl-2-(3-hydroxyisoamyl)-p-benzoquinon-3-yl] ethane (5), and 20.3% of 2-(3-hydroxyisoamyl)-3, 5, 6-trimethyl-p-benzoquinone (6), while that of 6-hydroxy-2, 2, 5, 8-tetramethylchroman (9) gave 21.2% of 6-hydroxy-2, 2, 5, 8-tetramethyl-7-(2, 2, 5, 8-tetramethyl-6-chromanyloxy) chroman (10) and 8.9% of 3, 6-dimethyl-2-(3-hydroxyisoamyl)-p-benzoquinone (11). The reaction of 6-hydroxy-2, 2, 7, 8-tetramethylchroman (12) in methanol afforded 49.6% of 2, 2, 7, 8-tetramethyl-5, 6-chromandione (13), 5.3% of 8-methoxymethyl-2, 2, 7-trimethyl-5, 6-chromandione (14), and 6.2% of 1, 2-bis (2, 2, 7-trimethyl-5, 6-chromandion-8-yl) ethane (15). The structure of 13 and 14 was determined by deriving them to phenazine derivatives, 2, 3-dihydro-3, 3, 5, 6-tetramethyl-1H-pyrano [3, 2-a] phenazine (16) and 2, 3-dihydro-5-methoxymethyl-3, 3, 6-trimethyl-1H-pyrano [3, 2-a] phenazine (17), respectively. The reaction of 12 in benzene or dimethylformamide gave 6-hydroxy-2, 2, 7, 8-tetramethyl-5-(2, 2, 7, 8-tetramethyl-6-chromanyloxy) chroman (18) (28.6% from benzene, 13.8% from dimethylformamide), 6-hydroxy-5-(6-hydroxy-2, 2, 7, 8-tetramethylchroman-5-yl)-2, 2, 7, 8-tetramethylchroman (19) (15.2 and 3.5%), 13 (24.3 and 40.9%), and 15 (1.9 and 10.0%). Oxidation of 6-hydroxy-2, 2, 8-trimethylchroman (21) gave 4.8% of 6-hydroxy-2, 2, 8-trimethyl-5-(2, 2, 8-trimethyl-6-chromanyloxy) chroman (22), 4.8% of 6-hydroxy-5-(6-hydroxy-2, 2, 8-trimethylchroman-5-yl)-2, 2, 8-trimethylchroman (23), 2.7% of trimer (24), and 19.7% of 2, 2, 8-trimethyl-5, 6-chromandione (25). The structure of 25 was determined by deriving it to the phenazine derivative, 2, 3-dihydro-3, 3, 5-trimethyl-1H-pyrano [3, 2-a] phenazine (26). The reaction of 2, 2-dimethyl-6-hydroxychroman (27) afforded 3.5% of 5-(2, 2-dimethyl-6-chromanyloxy)-6-hydroxychroman (28), 2.1% of 2, 2-dimethyl-5-(2, 2-dimethyl-6-chromanyloxy)-6, 7-chromandione (29), and 1.3% of 2, 2-dimethyl-6-hydroxy-5-(6-hydroxy-2, 2-dimethylchroman-5-yl) chroman (30). Thus, oxidation with 1 resulted in the simultaneous progress of introduction of oxygen and carbon-oxygen or carbon-carbon coupling.
Combined effect of drugs of animal origin (musk, bezoar, bufotalis) was examined by Boyden's method, using guinea pig polymorphonuclear leucocytes for inhibitory effect on leucocyte migration. A migration-inhibitory activity was found in musk, bezoar, ninhydrin reaction-positive substances musk peptide fraction (MP) extracted from musk, and smooth muscle contracting substances extracted from bezoar. MP was the most effective of substances tested. The active component of MP was determined to be a peptide from the fact that the activity was lost by tryptic hydrolysis and that the molecular weight was estimated to be about 1000, using ultrafiltration and Sephadex. The combined action of musk-bezoar was an antagonistic type. That of muskbufotalis was not observed but that of bezoar-bufotalis was a synergistic type. The apparent combined action of muskbezoar-bufotalis was not observed but interactions described above seem at least to be included.
1, 4, 5, 6-Tetrahydropyrrolo [3, 4-b] pyridin-7-ones (III) and 2, 5, 5, 7, 7-pentamethyl-1, 4, 5, 7-tetrahydrofuro [3, 4-b] pyridines (XII) were synthesized from 4-benzylidene-2, 3-dioxopyrrolidines (I) or 4-benzylidene-2, 2, 5, 5-tetramethyltetrahydrofuran-3-ones (XI), respectively, to find new cardiovascular drugs. Their 1, 4-dihydropyridine structures were confirmed from spectral data and by oxidation to the corresponding pyridines. It was also found that Madhav's intermediate was not 1, 4-dihydropyridine (III) but 4-(3-amino-2-ethoxycarbonyl-1-phenyl-2-butenyl)-2, 3-dioxopyrrolidine (II1). 1, 4, 5, 6-Tetrahydropyrrolo [3, 4-b]-pyridin-7-ones possessing 6-ω-aminoalkyl substituent (VII) were obtained from 6-ω-hydroxyalkyl derivatives (V) by the usual way. These compounds were tested for coronary vasodilating and antihypertensive activities, and several of them showed a marked coronary vasodilating activity.
Potentiation of β-adrenergic responses by musk, an animal-origin drug, has been studied mainly in cat papillary muscle preparations driven by electric stimulation (10 V, 0.8 msec, 0.4 Hz). In order to isolate the β-adrenergic potentiation factors, musk (20 g) was extracted and the potentiation activity of each fraction was assayed, using the cardiac muscle. The resultant fraction W5-a (64 mg) was homogeneous as judged by thin-layer chromatography. Dialyzed W5-a was soluble in ethanol and water but not in ether. The β-adrenergic potentiation of the cardiac muscle induced by W5-a was not due to the inhibition of catechol-O-methyl transferase (COMT) from the following facts : Parallelism between the COMT-inhibition activity and the β-adrenergic potentiation activity was not observed among the lots of musk tested, a purified musk fraction was a weaker inhibitor of COMT activity than the crude, and musk induced a potentiation of isoproterenol (IsP) to the same extent even in the high doses by which the inhibitory effect of pronethalol was surmounted. On the other hand it was observed that the purified fraction did not potentiate IsP-induced relaxation of smooth muscle preparations of trachea and taenia coli in guinea pigs. The crude musk itself potentiated the effect of IsP but not that of salbutamol which is well known as a COMT-resistant β-agonist. To conclude, the β-adrenergic potentiation induced by W5-a in musk was very selective to cardiac muscle, and also there may be other factors in musk which induce the β-adrenergic potentiation in tracheal muscle, at least in part due to the COMT-inhibitory activity.
The mean diameter, dm, of aggregated kaolin particles was measured by the sedimentation balance method in aqueous solution of various concentrations of chondroitin sulfate (Ch.) and/or of simple salt. It was found that dm decreases with increasing concentration of Ch. or of simple salt, because the adsorption of Ch. on kaolin disrupts the card-house structure of kaolin particles and because a simple salt shields the electrostatic attractive force between the face (negatively charged) and the edge (positively charged) of kaolin particles at low concentration of the salt, although the dm of kaolin particles becomes large at sufficiently high concentration of the simple salt by aggregation of kaolin particles. The dm is also affected by the sequence of mixing of Ch. and simple salt ; the dm is smaller in the case Ch. is added before simple salt, than when added after simple salt. The mean diameter, dm, of kaolin particles increases with pH when adjusted with Ca(OH)2, but decreases with pH when adjusted with NaOH. It was found that the amount of calcium ion adsorbed increases with pH. It was, therefore, concluded that calcium ions bind kaolin particles face-to-face by the adsorption of calcium ions on the face of kaolin. It was also found that OH ions are adsorbed on kaolin particles, and then pH of the suspension decreases to some value, depending on both the concentration and species of the salt added. This may be called a buffer action of kaolin.
A method for the quality estimation of musk in Japanese Pharmacopoeia, based on the determination of characteristic components, was investigated. By preparative thin-layer chromatography of ether extract, fractions containing muscone and a group of C19-steroids were separated and gas chromatographically analyzed. For the determination of muscone, cyclopentadecanone was found to be an excellent internal standard. The content of C19-steroids was determined after chromic acid oxidation leading to androstane-3, 17-diones. Application of the present method to the samples from various sources indicated it was most reliable for quality classification.
Ion-exchange celluloses and resins, silicic acids, cellulose fibers, and cotton wool were tested as adsorbents for leukocytes and platelets in whole blood and packed red cell suspension. Among the adsorbents tested, KC-Flock W-50S, Avicel PH101, and cotton from Gossypium barbadense were effective for removing leukocytes and platelets. When 3 column volume of blood was passed through a column packed with cellulose fibers, KC-Flock W-50S, 97% of leucocytes and 88% of platelets were removed. Recovery of red cells was quantitative. Efficacy of various adsorbents tested is discussed. Roughness and large surface area seem to be requisite factors for effective adhesion of leukocytes and platelets.
In order to examine the characteristics of intestinal absorption of stannic chloride, an in vitro experiment was carried out by circulating the reagent through segments of everted rat small intestine. The disappearance of tin from the mucosal solutions of rat duodenum, jejunum, and ileum followed apparent first-order kinetics. Tin absorption was approximately proportional to the concentration of tin in the mucosal solution and no difference was observed in the absorptions by rat duodenum, jejunum, and ileum. These results suggested that the absorption of tin from the rat small intestine occurs by passive diffusion. The effect of various food components such as organic acids, amino acids, and proteins on tin absorption from the rat intenstine and on its uptake by the intestine was also studied by the use of the small intestine in vitro and in vivo. Organic acids such as citric acid, tartaric acid, malic acid, and ascorbic acid significantly enhanced the tin absorption in vitro and in vivo. Tin uptake greatly decreased in the presence of these acids. Amino acids such as L-histidine, L-proline, and L-glutamic acid, and proteins such as ovalbumin and gelatin had little effect on tin absorption in vitro and in vivo. In addition, it was suggested that the effect of citric acid on the intestinal absorption of tin was due to a modification of intestinal permeability and the enhanced permeation of tin across the intestine in the form of tin-citric acid complex.
The reaction of 3, 3-diacetyl-1-acyltriphenylphosphonium allylides (4) with bases gave 5-substituted 2-acetyl-4-triphenylphosphoniophoniophenolates (5) and 5-substituted 4-acetyl-2-triphenylphosphoniophenolates (6). These structures were established by chemical and ultraviolet spectral methods. On the other hand, deacetylation occurred by the treatment of 4 with 10% hydrochloric acid to give 3-acetyl-1-acyltriphenylphosphonium allylides (18). 3, 3-Diacetyl-1-benzoyltriphenylpuhsphonium allylide (4a) was pyrolyzed to 1, 1-diacetyl-4-phenylbutenyne (17).
A new apparatus to measure the extra-weak chemiluminescence based on single photo-electron counting method was constructed and some working conditions for this apparatus were examined experimentally. The upper limit of the intensity of input light for quantitative determination in this apparatus is about 10-12 W and the lower one is about 6×10-16 W. Luminol was determined by the use the apparatus in 0.1 pg order of the magnitude by its oxidative reaction system. Small quantities of ATP (10-13 mol) and NADH (10-13 mol) were also determined in the firefly and bacterial luciferase system, respectively.
An attempt was made to measure the extra-weak chemiluminescence from aqueous solutions of various drugs by the apparatus newly constructed. From the solutions of comparatively unstable drugs such as L-ascorbic acid, pyridoxal-5-phosphate, etc., emission of extra-weak light was observed, and no luminescence was observed in solutions of comparatively stable drugs such as succinic acid, glycine, etc.
After oral administration of 5-fluoro-1, 3-bis (tetrahydro-2-furanyl)-2, 4-pyrimidinedione (FD-1), the level of 5-fluoro-2, 4-pyrimidinedione (5-FU) was 5 to 7 times higher in the plasma and normal tissues and 8 to 12 times in tumor tissue than after administration of 5-fluoro-1-(tetrahydro-2-furanyl)-2, 4-pyrimidinedione (FT). Moreover, these levels were maintained longer than after administration of FT. In tumor tissue, the concentration of 5-FU was still as high as 1.42 μg/g 12 hr after administration of FD-1. FD-1 was degraded to 5-fluoro-3-(tetrahydro-2-furanyl)-2, 4-pyrimidinedione (3-FT) by liver microsomal drug-metabolizing enzymes in vitro and to FT spontaneously. Subsequently, FT was converted enzymically to the active substance, 5-FU, and 3-FT changed to 5-FU spontaneously. Conversion of FD-1 to 5-FU via 3-FT was greater than via FT. It is concluded that a large amount of 5-FU formed after administration of FD-1 is formed via 3-FT. γ-Hydroxybutyric acid was found to be formed in vivo and in vitro from the tetrahydrofuranyl group of FD-1.
It was confirmed from spectrophotometric method and solubility method that there is an interaction between benzoic acids and caffeine compounds in an aqueous solution, and a constant (K) for complex formation was calculated by the solubility method. In order to examine the force of interaction between benzoic acids and caffeine compounds, electronic property was analyzed by the Pariser-Parr-Pople method from quantum chemical standpoint. There was a correlation between the size of complex formation constant and the lowest empty orbital energy level (εlv) of benzoic acids, assum ing that benzoic acids had an electron affinity and caffeine compounds at electron-donating property. Phenacetin has a large electron affinity, and its complexasation showed no correlation between εh0 of benzoic acids and complex formation constant. Caffeine compounds are molecules with a planarity, and there was a correlation between K and the size of molecular plane, suggesting the participation of a hydrophobic combination. From these evidences, interaction between benzoic acids and caffeine compounds seemed to be a complex mechanism in which charge transfer and hydrophobic binding participate.
In order to examine fragrant components, the essential oil of the flowers of Paulownia tomentosa STEUDEL was examined and, as the odor substances, hydroquinone dimethyl ether, 1-octen-3-ol, cis-3-hexen-1-ol, l-linalool, benzyl alcohol, phenethyl alcohol, benzaldehyde, anisaldehyde, ethyl palmitate, phenol, cresol (o-, m-, p-), 3, 4-dimethoxyphenol, and 12 kinds of volatile acid were identified. Apigenin, ursolic acid, β-sitosterol, campesterol, stigmasterol, β-sitosterol-β-D-glucoside, caffeic acid, p-hydroxybenzoic acid, p-coumaric acid, 3 kinds of fatty acid, and 17 kinds of normal paraffine were isolated from the steam distillation residue, as odorless components.
Fifteen strains of Actinomycetes were isolated from water and bottom sediment of Lake Tairo and their metabolites were examined by gas chromatography-mass spectrometry. The Actinomycetes were cultured on two different media, yeast-malt extract agar and oatmeal agar, at 28°for 14 days and the caltured media were subjected to steam distillation to recover volatile metabolites. 2-Methylisoborneol and geosmin were found in almost all the cultured media. 2-Phenylethanol was identified in the cultured solution of S. phaeofaciens, S. lavendulae, S. versipellis, and S. neyagawaensis. It was also noted that salicylaldehyde, phenol, and 3-octanone were biosynthesized by S. lavendulae, S. phaeofacience, and S. chibaensis, respectively.
The conformation of calcium ketyl radicals derived from (1-naphthalenyl) phenylmethanone and (2-methyl-1-naphthalenyl) phenylmethanone was examined from electron spin resonance spectra. The phenyl and carbonyl groups were coplaner and the twisting angle about the C-C bond connecting naphthyl and carbonyl was found to be 26°at room temperature and 46°at -20°. The twisting angle of the radical was smaller than that of a neutral molecule.
Effect on the leucocytosis-promoting activity of Parotin was examined by incubation of Parotin with rabbit serum or organ homogenate at 37°for 60 min, and the following facts were found. 1) The leucocytosis-promoting activity of Parotin was inactivated by incubation with rabbit serum or with homogenate of the bile, liver, or spleen. 2) This activity was not affected by incubation with the homogenate of the heart, kidneys, adrenal, thymus, thyroid, testis, sublingual gland, parotid gland, or submaxillary gland.