Moisture absorption and heat stability were examined with magnesium oxide, heavy magnesium oxide, desiccated aluminum hydroxide gel, and synthetic aluminum hydroxide, all in the Japanese Pharmacopoeia, by storage over a long period of time in 5-90% relative humidity. It was thereby found that these samples are comparatively stable below 60% of relative humidity and that majority of the samples showed a two-step decrease in weight by heating. This two-step decrease was found to be the liberation of adsorbed water molecule alone in the first step and that of water molecule realted to the structure in the second step. This fact agreed with the result of X ray diffraction data. Based on the above data, chemical kinetic analyses were made according to the method of Freeman and Carroll, and activation energy and reaction order were calculated.
A new flavone glucoside, named unranin and linarin, were obtained from the flowers of Linaria japonica MIQ. and the structure of unranin was assumed to be 7-rhamnoglucoside or 7-glucorhamnoside of 4'-O-methylscutellarein.
A conventional coulometric titration method for the determination of a small amount of Sulfamethizole, Sulfaphenazole, and Sulfamethoxypyridazine (listed in the Pharmacopoeia of Japan, 8th Ed., 1971) was examined. Sulfamethizole, Sulfaphenazole, and Sulfamethoxypyridazine were brominated quantitatively with electrolytically generated bromine, in an aqueous hydrochloric acid-potassium bromide medium. The end point was detected by a potentiometer. Sulfamethizole and sulfamethoxypyridazine reacted with 2 moles of bromine and changed to their dibromides, 2-(3, 5-dibromosulfanilamido)-5-methyl-1, 3, 4-thiadiazole and 3-(3, 5-dibromosulfanilamido)-6-methoxypyridazine, respectively, while sulfaphenazole reacted with 3 moles of bromine and changed to its tribromide, 5-(3, 5-dibromosulfanilamido)-4-bromo-1-phenylpyrazole. The brominated position of these sulfonamides was checked by infrared and nuclearmagnetic resonance spectrograms.
1. Ultraviolet irradiation of thyroxine, diiodotyrosine, and their analogous compounds was carried out to elcidate the process of photodegradation of thyroxine by ultraviolet ray. 2. For separation and detection of the degraded products, measurement of ultraviolet and infrared absorption spectra, and paper chromatography, thin-layer chromatography, and column chromatography were carried out. 3. Experimental results revealed that the irradiation of ultraviolet ray to thyroxine liberated iodine at first and then cleaved the ether bond in the molecule, and liberated α-phenyl ring from thyroxine. Triiodothyronine, diiodotyrosine, monoiodotyrosine, tyrosine, phenylalanine, serine, alanine, and glycine were detected as the photodegradation products. 4. Production of diiodotyrosine from ultraviolet ray irradiated thyroxine was confirmed by the measurement of infrared absorption spectrum. 5. The fact that thyronine or tyrosine which lacked iodine in the molecule was more resistant than iodoamino acids to ultraviolet ray irradiation suggested that instability of thyroxine and related compounds to ultraviolet ray was due to the liberation of iodine from these compounds.
Preparation of d-cis-3-acetoxy-5-[2-(dimethylamino)ethyl]-2, 3-dihydro-2-(p-methoxyphenyl)-1, 5-benzothiazepin-4(5H)-one hydrochloride (IIId) with potent coronary vasodilatory activity is described. Attempted resolution of III, IX, and VI with various optically active acids was unsuccessful. VIIId was barely obtained by numerous repeated recrystallization of d-10-camphorsulfonate of VIII in a low yield. On the other hand, resolution of V with cinchonidine was carried out successfully. VIl was also obtained by resolution of VI with ephedrine. IIId and IIIl were easily prepared from Vd and Vl, respectively.
Two new phenolic compounds, glycoside A, C16H22O7, mp 110-112°, [α]20D-52.1°, and glycoside B, C20H22O8, mp 235-238°, [α]20D-72.0°, were isolated from Rheum rhizome. Structures of these substances were determined by nuclear magnetic resonance and mass spectra.
In order to clarify the ability of complex formation and solubilizing action of acid amides, quantum chemical approach using semi-empirical Pariser-Parr-Pople method was made and the following results were obtained. 1) On the basis of molecular orbital considerations, it was expected that acid amides act as electron acceptors and the solubilizates act as electron donors. 2) Formation constants of complexes had a linear relationship with energy levels of the lowest-vacant molecular orbitals (εlv) of acid amides. These results calculated by using the Pariser-Parr-Popple method agreed well with those obtained by Huckel method. The Huckel method was used to elucidate the structure of solubilizer-solubilizate complexes, and stabilization energy obtained from the formation of complexes was calculated by using the purturbation method. It was recognized that the formation constant of complexes had a linear relationship with the stabilization energy, and the correlation coefficient was 0.87.
To study enzymological properties and usefulness of continuous enzymic reaction of the insoluble derivatives of semi-alkaline proteinase (SAP) from Aspergillus melleus, the purified SAP was transformed into insoluble forms by coupling its native form to insoluble carriers, using CM-cellulose (CMC), copolymer of ethylene and maleic anhydride, (EMA) and Sephadex. The optimum temperature and thermostability increased by the insolublization. The optimum pH of EMA-SAP and CMC-SAP was at pH 7-10 and 10-11, respectively, against soluble SAP and Sephadex-SAP at pH 8.0. The effect of protein denaturation reagents, metal ions, and surface-active reagents on insoluble SAP was also examined. Insoluble SAP preparations were little inhibited by sodium dioctylsulfosuccinate whereas soluble SAP lost most of the activity. The activity remaining in the insoluble SAP stored in a buffer of pH 6.0 for several months was much higher than that of soluble SAP and a column of Sephadex-SAP through which casein solution was continuously passed for 20 days retained about 40% of initial activity. Little difference between the soluble and insoluble SAP with respect to Km values and the energy of activation suggested that the conformation of SAP does not change by insolublization. From the analysis of amino acid composition of Sephadex-SAP, the basic amino groups of SAP useful for coupling with activated Sephadex were lysyl and arginyl residues.
The metabolic fate of 5-fluorouracil was studied in the rat by using 14C-labeled compound in ointment. Following the percutaneous administration of 5-fluorouracil in ointment, the rat excreted about 4% of the administered radioactivity in urine, 0.2% in feces, and 2.7% in respiratory CO2 within 72 hr, and about 90% of administered radioactivity remained at the site of application. Absorption of 5-fluorouracil in ointment from the skin was almost completed within the first 1 hr after ointment application and the maximum levels of radioactivity in rat tissues and blood also reached in about 1 hr after the application. Distribution of radioactivity in rat tissues was more highly located in the liver, kidneys, and in dermic and hypodermic tissues than in other tissues. After application of 0.5g per rat, of ointment containing radioactive 5-fluorouracil, the radioactive substances excreted in urine after 72 hr were unchanged 5-fluorouracil, α-fluoro-β-ureidopropionic acid, α-fluoro-β-guanidopropionic acid, and α-fluoro-β-alanine which were present in about 7.9%, 13.5%, 5.8%, 66.1%, and 3.7% of total radioactivity in the urine, respectively.
1) An in vivo study was made on the effect of ascorbic acid and ferrous ion on the formation of lipid peroxide in the rat liver mitochondria. Lipid peroxide content increased 24 hr after the intraperitoneal administration of ascorbic acid (176 mg/kg/day) and ferrous chloride (28.2 mg Fe2+ kg/day). In addition, Iipid peroxide content increased by about two fold after daily administration of ascorbic acid and ferrous ion for 1 week. 2) The amount of unsaturated fatty acids (C18 : 2, C20 : 4) in mitochondria decreased during increase in the formation of lipid peroxide. Among the composition of phosphatidylserine and phosphatidylinositol, diminution of linoleic acid and arachidonic acid was predominant. 3) Total sulfhydryl content in mitochondria decreased markedly by the application of ascorbic acid and ferrous ion.
An octapeptide fragment containing the heme moiety hemeoctapeptide (CHP) was prepared by peptic and tryptic digestion of yeast cytochrome c (Candida krusei). CHP was highly purified by chromatography and other methods by which any impurity due to pyrogenic and antigenic substances was completely excluded. Two different types of experimental hypoxia were employed in which a protective effect of CHP and its related compounds against the denaturation of cells and organelles due to oxygen deficiency was examined. First, hemorrhagic lipemia caused by hemic hypoxia was produced in rabbits by a consecutive bleeding for several days' duration. The animals which were given cytochrome c or CUP (1 mg/kg, daily, as a form of saline solution) intravenously showed less elevation of serum lipid levels than control animals treated with an equivalent volume of saline. Second, anoxic hypoxia was produced by the exposure of rats to simulated high altitude, which resulted in the disruption of lysosomes and in release of tissue enzymes such as acid phosphatase and transaminases into the circulatory system. In each experiment, 6 or 8 rats, which were injected with saline or drug solution intraperitoneally 15 minutes before the exposure, were placed in a well ventilated vacuum box and the atomospheric pressure was reduced and kept constant at the 300 Torr during the test period. Such changes in both tissue and serum enzyme levels were fairly well suppressed in the aimals pretreated with CHP (1 mg/kg). In addition to these biological data, histopathological and histochemical observations showed that tissue damages due to high altitude which were most apparent in the liver were limited when the animals were given CHP. It is suggested that the physiological function of externally given cytochrome c may be attributed to the biological activity of an effective intermediate such as CHP.
Pyrolysis gas chromatography was applied for the analysis of (35 species of) natural resins. Benzene, toluene, xylene, styrene, and light gases (CO, CO2, CH4, C2H6, C2H4) were identified as their pyrolysis products at 800°. Siam Benzoin produced much benzene, while Sumatra Benzoin, Storax, and Peru Balsam yielded much styrene. Amber, Dammar, and Copal produced much toluene and xylene besides benzene. Rosins and gum resins showed less, and plant gums showed the least production of these compounds. Natural resins were thus classified according to their pyrograms.
Pyrolysis gas chromatography at 450° was investigated for the identification of natural resins. Each natural resin tested gave different pyrogram, and the characteristic peaks were identified as follows : From Copal, m- and p-xylene, and limonene, acetone from Rosins, C9H14 and isoprene from Gamboge, ethyl acetate from Guaiacum, methanol from Dragon's Blood, toluene and styrene from Storax and Peru Balsam, and monoterpene compounds from Elemi and Canada Balsam. Consequently, it was concluded that natural resins can be identified by their characteristic pyrogram at 450°.
Non-steroidal anti-inflammatory drugs and α-tocopherol were tested in vitro for inhibition against hemolysis and lipid peroxidation induced by H2O2 in erythrocytes from healthy mongrel dogs. In concentration over the range of 10-6 to 4.5×10-4M, anti-inflammatory drugs inhibited both hemolysis and lipid peroxidation, while α-tocopherol had no effect. Against peroxidation of free unsaturated fatty acids caused by various means (H2O2, UV irradiation, ascorbic acid+Fe+2, and hemoglobin), antioxidants including α-tocopherol showed a marked inhibition, whereas almost all anti-inflammatory drugs had no effect. H2O2 was not destroyed directly by both antioxidants and anti-inflammatory drugs. Diphenyl-p-picrylhydrazyl, a model compound of free radical, was reduced powerfully with antioxidants but not with anti-inflammatory drugs.
The effect of particle size on ζ-potential was studied by the streaming potential method. The streaming potential was measured with varying particle size of untreated glass particles and glass particles coated with dodecylamine in sodium chloride or dodecylammonium chloride solution. These experiments showed that |ζ| became larger, the smaller the particle size was. Necessary corrections have to be made to obtain a reliable value of ζ-potential. The corrections for surface conductance, overlapping of electrical double layer, and radius effect were proposed by many authors experimentally and theoretically, but these do not seem to give reliable values. A corrected equation [numerical formula] was derived here, theoretically, with consideration of the tortuosity of liquid flow, qη, and the tortuosity of ion flux, qκ, but a reasonable value was not obtained.
Previous studies on the photodynamic inactivation of various biological substances in the presence of riboflavin have shown that photoinactivation of yeast alcohol dyhydrogenase (EC 220.127.116.11) and Bacillus subtilis transforming DNA was greatly accelerated by the addition of adenine. Attempts were made to examine whether a similar phenomenon would also be observed in the riboflavin-sensitized photoinactivation of Escherichia coli tRNA and to elucidate the basic mechanism involved in the reaction. The riboflavin-sensitized photoinactivation of tRNA is remarkably accelerated by the addition of adenine under aerobic conditions. Adenine can also promote the photodynamic inactivation of tRNA with FAD, FMN, and lumiflavin, while the reaction with lumichrome, Methylene Blue, thiopyronine, and Eosin Yellow is not affected by adenine. The adenine derivatives such as adenine 1-N-oxide, AMP, ADP, and NAD also accelerate the riboflavin-sensitized photoinactivation of tRNA, whereas guanine, cytosine, thymine, uracil, and adenosine do not accelerate the photoinactivation significantly. The enhancement of the riboflavin-sensitized photoinactivation of tRNA by adenine may be ascribed to the promotion of the destruction of guanine residues in tRNA and to the acceleration of changes of base stacking and the disruption of base pairs, which seems to result from the destruction of guanine residues in tRNA. The promotion of the photodynamic degradation of 4-thiouracil residues may also be responsible for the enhancement of the inactivation of tRNA, since disappearance of its characteristic absorption maximum near 335mμ was promoted by adenine. The mechanism through which adenine causes the enhancement of the riboflavin-sensitized photoinactivation of tRNA is discussed.
During the preceding studies on the rearrangement of allylic xanthate to allylic dithiolcarbonate, 3) it was sometimes observed that the product was contaminated with trithiocarbonates. The present study was undertaken to clarify the formation condition and the formation mechanism of trithiocarbonates. As a result, it was concluded that sodium 2-alkenyl trithiocarbonate, the precursor of di(2-alkenyl) and 2-alkenyl alkyltrithiocarbonates, might be formed by the following routes : Sodium 2-alkenyl dithiolcarbonate was formed by thermal rearrangement of sodium 2-alkenyl xanthogenate and then decomposed into sodium 2-alkenyl sulfide which reacted with carbon disulfide to give sodium 2-alkenyl trithiocarbonate. For the preparation of unsymmetric trithiocarbonates, it is recommended that alcohols are reacted with excess carbon disulfide in dimethyl sulfoxide containing powdered sodium hydroxide.
When the chemical shift of the methyl-proton and methoxyl-proton is plotted against the concentration of a mixed solution of water with 1-methoxy-2-propanol, methylcarbitol, carbitol, cellosolve, dipropylene glycol, isopropylcellosolve, or hexylene glycol as the solubilizing agent, shift to a higher magnetic field is observed at a certain concentration in any of the mixtures. The plot of the chemical shift against dielectric constant of the mixed solution showed a linear relation between dielectric constant at ca. 15-45 and the chemical shift of methyl-proton and methoxyl-proton. Consequently, these mixed solutions are considered to undergo association at this concentration. In mixed solutions of water with 1-methoxy-2-propanol and hexylene glycol, a change in spin coupling state is seen in the methylene-proton at a specific concentration and this concentration is that at which the foregoing methyl- and methoxyl-protons show a shift to a higher magnetic field, and the same concentration at which an inflexion appeared in the plot of dielectric constant with solubility and physical properties. This phenomenon is considered to result from the association of the mixed solutions at this concentration.
The present work was undertaken in order to synthesize oxazolo[4, 5-d]pyrimidines which are of interest as antimetabolites. The Schotten-Baumann reaction of 2, 4-diamino-5-hydroxy-6-methylpyrimidine (III) with ethyl chlorocarbonate gave 2, 4-diamino-5-ethoxycarbonyloxy-6-methylpyrimidine (XII). XII was refluxed in pyridine to give 5-amino-7-methyloxazolo[4, 5-d]pyrimidin-2(3H)-one (XIII), which was alkylated with alkyl halide to give 3-alkyloxazolo[4, 5-d]pyrimidine derivatives (XIVa-d). Also the Schotten-Baumann reaction of III with ethyl bromoacetate and ethyl α-bromopropionate gave pyrimido[5, 4-b][1, 4]oxazine derivatives (XV) and (XVII), respectively.
Seasonal variation in the chemical constituent of Artemisia feddei growing in Kochi Prefecture was examined and it was found that the components contained in the spring plant gradually change to those of the mature plant during the period from the middle of July to early August.
Zirconium formed a blue-violet complex with Bromopyrogallol Red in the presence of zephiramine in weakly hydrochloric acid solution. This complex showed an absorption maximum at around 639 nm, and the absorbance was constant over the acidity range of 0.18-0.28N (the final acidity). The proposed method covered a concentration range of 3-40 μg of zirconium in 25 ml of solution. The molar absorptivity was about 37000. The precisions in coefficient of variation were 2.0 and 1.2% for 10 and 30 μg, respectively, of zirconium. Many ions did not interfere with the experiment, but hafnium, vanadium, molybdenum, iron(III), mercury(II), fluoride, and phosphate interfered even when their amount was the same as that of zirconium. The permissible amount of iron (III) was increased by the addition of L-ascorbic acid.