Condensation of o-phthaldehydic acid and glycine anhydride in the presence of acetic anhydride and sodium acetate results in the formation of 3, 6-bis (o-carboxybenzal) dioxopioperazine (II), with 3, 6-diphthalidyldioxopiperazine (I), as a by-product. The chief condensate (II) decomposes into isocarbostyril-3-carboxylic acid when heated with sodium hydroxide and also forms this acid when boiled with hydriodic acid and red phosphorus, instead of the double bond alone being reduced. The by-product (I) decomposes into o-toluic acid and glycine when treated with hydriodic acid and red phosphorus and into isocarbostyril-3-carboxylic acid with sodium hydroxide. The same condensation of m-and p-phthalaldehydic acid as in the case of the ortho-compound results in the respective formation of bis (m-and p-carboxybenzal) dioxopiperazine which form m-and p-carboxyphenylalanine on being treated with hydriodic acid and red phosphorus.
Pharmacological action of kainic acid, the effective principle of Digenea, was further examined. Kainic acid does not inhibit the gill respiration of a clam (Corbicula leana Prime) or the respiration of the tissues of hog ascaris (Ascaris suilla), but inhibits the dehydrogenase action in the muscle. Injection of 1-2mg./kg. of kainic acid in rabbits inhibited heart motion, damaged the kidney and liver, and affected changes in the blood cell counts of the plasma. In examining the action of kainic acid on smooth muscles, it was found that the vomiting that appears on subcutaneous injection in puppies is the direct action of kainic acid on the smooth muscles and irritation of the vomiting center.
In order to generalize the new method for N-monoalkylation of β-aminoalcohols (Pharm. Bull. (Japan), 1, 325 (1953)). dl-ψ-5-phenyl-2, 4-dimethyloxazoline was synthesized by the treatment of N-acetyl-dl-ψ-norephedrine with conc. surfuric acid. The former was reacted with methylating agents to give the postulated dl-ψ-5-phenyl-2, 3, 4-trimeth-yloxazolinium salts as oily products which were very sensitive to moisture and rapidly added one mole of water to form the salt of O-acetyl-dl-ψ-ephedrine by retention. The oily product was boiled, immediately after formation, in dry acetic acid-sodium acetate, avoiding moisture, and gave O, N-diacetyl-dl-ephedrine by inversion. Thus it was found that the amine was not substituted by more than one alkyl group and that the retention or inversion of the configuration of the resultant compound could be controlled at will by the treatment either in wet or dry conditions.
L-Fucose was synthesized by deriving L-fucitol to 2:3, 4:5-diisopropylidene-L-fucitol, its oxidation to potassium 2:3, 4:5-diisopropylidene-L-fuconate with potassium permangante, hydrolyzed to L-fuconolactone, and by final reduction with sodium amalgam.
Although thienopyridines, the benzene portion of the isoquinoline ring substituted with thiophene, have been synthesized, the compound in which the benzene portion had been substituted with furan, i, e. furo [3, 2-c] pyridines have not been synthesized as yet and this was attempted. Furylnitroethylene was reduced with lithium aluminum hydride to β-2-furylethylamine which was condensed by the Schotten-Baumann method respectively with benzoyl, 3, 4-dimethoxybenzoyl, 3, 4-methylenedioxybenzoyl, and 3-methoxy-4-benzyloxybenzoyl chloride to the corresponding acid amides, and were finally cyclized by the Bischler-Napieralski reaction to the objective 1-aryl-3, 4-dihydrofuro [3, 2-c] pyridines.
Of the 6 kinds of theoretically available thienopyridines, several kinds of 1-aryl-3, 4-dihydrothieno [3, 2-c] pyridines were obtained by deriving 2-thienaldehyde to β-2-thienylethylamine to which were respectively condensed 3, 4-dimethoxy-, 3, 4-methylenedioxy-, and 3-methoxy-4-benzyloxy-benzoyl chloride to the amide, and finally cyclized to the objective compounds. Pharmacological action of three kinds of thienopyridines and furopyridines was examined and 1-(3′-methoxy-4′-benzyloxyphenyl)-3, 4-dihydrofuro [3, 2-c] pyridine was found to cause a marked intestinal contraction and inhibition of intestinal movements, and to possess oxytocic action about one-fiftieth that of Oxytocin.
Growth inhibition of Staphylococci (Terashima strain) was examined with 30 kinds of cyanine dyes, including aminovinyl, merocarbocyanine, methineoxonol, and benzal compounds. It was found that the aminovinyl and its allied series of dyes were effective while methineoxonol dyes also showed a slight growth inhibition. The aminovinyl compounds were also effective against coli bacilli. Effect of dyes on the tissue permeability was examined by mixing these dyes with black ink with or without hyaluronidase and injecting the fluid on rabbit's back and determining the size of the blackened area. It was thereby found that 5-(ω-benzoylvinyl)-thiobarbituric acid and methiodides of 2-ω-trichloromethylvinylpyridine and -quinoline gave good biological effect.
Reductive decomposition of folic acid with zinc powder and hydrochloric acid, followed by the azotometry for aromatic primary amines was found to make quantitative determination of folic acid possible.
The lignan, m.p. 146.5-147°, [α]24D: -87.3° (c=1.79 in CHCl3), were obtaind with wax from the dried leaves of cypress (Chamaecyparis obtusa). The liguan was also obtained from Chamaecyparis obtusa var. breviramea and corresponded to the molecular formula of C20H16O6. It is assumed to be identical with savinin obtained from the leaves of the juniper species by Hartwell and others, by the agreement of its specific rotation and ultraviolet and infrared absorption spectra. This substance is also identical with hibalactone obtained from Nikko cypress and Ch. obtusa var. breviramea and must be α-(3, 4-methyledioxybenzylidene)-β-(3, 4-methylenedioxybenzyl)-butyrolactone (I) from the ultraviolet and infrared spectral data.
When the mono- and di-salt of the ar -N-oxide of quinine or dihydroquinine is warmed with phosphoryl chloride in chloroform, 2′-chloroquinine or -dihydroquinine is formed, as in the case of the free base. Mono salt gives a better result and the monothiocyanate, an intermediate in the synthesis of the N-oxide of quina base, is the most suitable since it gives the 2′-chloro compound in about 90% yield. Application of sodium methoxide, phenoxide, methanethioid, and piperidine to 2′-chloroquinine and -dihydroquinine yields the corresponding 2′-methoxy, -phenoxy, -thiomethyl, and N′-piperidyl derivatives.
By the reaction of aromatic nitro compounds and α-picoline, in the presence of sulfur, p-phenoxyanilide, p-allyloxyanilide, and p-isopropoxyanilide of thiopicolinic acid and their corresponding benzothiazole derivatives were obtained. In the case of aromatic primary amines, only the p-anisidide and p-phenetidide of thiopicolinic acid were obtained and not their corresponding benzothiazole derivatives that they were submitted separately to oxidative cyclization to the benzothiazoles. It has experimentally been proved, through the use of nitrobenzene, that the aromatic nitro radical acts as an oxidative cyclization agent. Thiopicolinic acid p-anisidide was found to be effective in antibacterial tests in the following dilutions: Tubercle bacilli (judged after 4 weeks, in Kirchner medium) 320, 000; Staph. aureus, <10, 000; Esch. coli <10, 000; S. dysenteriae (Shiga), <10, 000; Vibrio cholerae <10, 000.
Using spots developed on a filter paper from 24 kinds of tar dyes for foodstuff, reflect-ance spectra in the visible range were measured with Beckman Spectrophotometer Model B, with a diffuse reflectance attachment. The majority of the spectra thereby obtained were found to agree well with absorption spectra of these dye solutions. A mixture of several kinds was submitted to paper chromatography and separated into componental parts. In order to determine the composition of such dye mixtures the separated colored spots were submitted to reflectance measurement. By comparing these measured values with the reflectance of the authentic dyes, satisfactory agreement was obtained.
Thirty-two kinds of 2, 4-dinitrophenylhydrazones were prepared of three aliphatic and eight terpenoid carbonyl compounds, furfural, 5-nitrofurfural, eleven aromatic aldehydes, and eight aromatic ketones. Their surface color was compared by photo-reflectance measured in the range of 400-700mμ with the Beckman Spectrophotometer with diffuse attachment. From the data obtained, reflectance ratio in the different wave lengths and saturated reflectance were compared. By comparing the characteristics of the inclination and the wave length at the standing point of the reflectance curves, it was found possible to represent a delicate difference in the color tone.
From the data of spectroreflectance, the surface color of 30 kinds of 2, 4-dinitrophenyl-hydrazones was plotted on the C. I. E. color chart. By this operation, x and y values, brightness (Y), dominant wave length, and purity of each color could be determined.
Absorption maxima of the 32 kinds of 2, 4-dinitrophenylhydrazones in acetone solution were measured. The absorption maxima were also compared on the addition of alkaline reagent, such as 1% ethanolic sodium hydroxide, pyridine, or ammonia water. Some observations were made as to the relationship between characteristic color tone and chemical structure.
Using Staphylococcus aureus 209P, uptake of arginine into the free amino acid pool in bacterial cell was examined. It was found that arginine in the medium is not taken up at all at 0° but is taken up at 37°, and that arginine uptake is influenced by the kind and pH of the buffer solution used for the medium. The uptake was the best when a phosphate buffer of pH 7.4 was used, followed by the Veronal buffer of pH 7.4 and glycine buffer of pH 9.9 The arginine uptake was considerably accelerated by the addition of glucose or phosphate to the glycine buffer than the glycine buffer alone. However, in the case of Veronal buffer, glucose was found to act inhibitively and the reason for it is still obscure, although the fact was also noted by Melchior. Canavanine, an antagonist of arginine, also suppressed arginine uptake but such effect was not seen in lysine and other amino acids.
Canavanine is used in biological experiments as an antimetabolite of arginine but suitable method has not been found for separatory detection of these two substances in a sample. It has been found that good results can be obtained in this direction by coloration after separation of the two substances by paper electrophoresis in Veronal buffer (pH 8.6) or by buffered chromatography with phenol and borate buffer (pH 9.3). The colorimetric determination of canavanine in the presence of arginine was found to be impossible in a biological system because of an obstruction by metabolites but a few knowledge were gained on the determination of arginine in the presence of canavanine. For example, the determination of arginine by the Sakaguchi reaction can be freed from the influence of canavanine if hypobromite is used but coloration is interfered by the presence of a reductive substances such as glucose. Microbiological assay using Streptococcus faecalis R is not affected by the growth inhibition of canavanine nor by other arginine homologs but stimulation of growth by canavanine appears and the determination values obtained are only relative. Separation of canavanine and argininewwas effected by the use of an jon exchanger, Amberlite IRC-50 or IRA-400.
Synthesis of glutamylcholine by the condensation of N-carbobenzoxy L-glutamic anhydride (I) and choline chloride (II) followed by reduction, was found to be accompanied by a minute amount of a by-product which was found to contain L-2-oxopyrrolidine 5-carboxylic acid choline ester (III) and L-glutamyldicholine (VII), confirmed with the synthetic products.
One of the active components of Cruculon (Takeda), a curcuma preparation, is said to be ferulic acid and its quantitative determination has already been reported. Having observed that the vanillic acid present as a contaminant in the ferulic acid hinders the colorimetric determination of the latter the separation of both compounds by paper chromatography was carried out and the separated ferulic acid was colorimetrically determined by diazotized sulfanilic acid solution. So far as the authors' experiments are concerned, the most suitable developing solvent for the paper chromatographic separation is 2.5% aqueous solution of boric acid.
The colorimetric determination of santonin, using sodium methoxide solution, was examined by the stochastic means. The largest factor that gives variation in error was found to be the heterogeneity of the sodium methoxide solution. It seemed that the atmospheric moisture also affects this variation that it seemed inadvisable to carry out this determination on a rainy day.
Santonin content in Artemisia kurramensis Quaz. can be determined by extracting the plant material with barium hydroxide, acidification of the baryta extract with hydrochloric acid, shaken with chloroform, and purifying the finally obtained chloroform solution of santonin, or chloroform extract of this plant itself by passage through an adsorption column filled with activated alumina or carbon, and finally by colorimetric determination with sodium methoxide. The sample required for this method is 0.1g. and the procedures are so simple that it can be adopted for the detection of the variation of santonin content during growth or in various parts of the plant body.
It had been found that the relation Y=39.6-43X holds for the Rb value (X) and tannic acid content (Y) of the leaves of Geranium nepalense. Based on these finding, Rb values of the plants collected throughout Japan during July and August were studied and following points were clarified. 1) Rb values are generally 0.29-0.56 and from the above equation, the tannin content is calculated as 27-16%. 2) Mean values of plants collected during July were approximately equal to those in August. 3) Rb values of plants bearing white flowers were smaller than those of red flowers while average values from the same species collected during July were approximately equal to those in August. 4) Taking the Rb value of the products meeting pharmacopoeial standards as below 0.45, or taking 0.47 by considering experimental errors, 79% of 107 samples were found to meet this standard. If the tannic acid content is taken as over 20%, Rb value will become 0.46, and 73% meets the standard. Summarizing the series of studies reported in Parts I to V of this series, following new method of examination is proposed. Leaves are plucked uniformly from the sample, dried at 80° for 30 minutes, and reduced to a pharmacopoeial coarse powder. To 40cc. of boiling distilled water, 0.4g. of this coarse powder is added, and refluxed for 30 minutes, under brisk boiling. This is cooled to the room temperature, 5cc. of the supernatant is added to 15cc. of distilled water, and this solution is divided into four portions. To each portion, molar eqivalent of sodium hydroxide is added, extemporaneously, 5cc. of 0.2% aqueous solution of methylene blue is mixed in with agitation, and this mixture is transferred equally to eight glass cylinders of 3cm. internal diameter and height. Two standards of 5cc. of the foregoing 0.1% methylene blue solution in identical glass cyclinder are placed in the middle, with the sample solutions on the either side, and the capillary image is developed by suspending strips of filter paper (Toyo Roshi No. 50, 2×30cm.), dipping the end in each cylinder. The average height (cm.) of methylene blue in the mixture is designated as B, that of the standard as A, and the ratio of B to A, B/A, must be less than 0.46.
Cleavage reaction of the biscoclaurine-type alkaloids, O-methyldauricine (I) and isotetrandrine (III), was carried out with the Grignard reagent, cobalt chloride as the catalyst, and tetrahydrofuran of dimethylaniline as a solvent. (I) was recovered on carrying out the reaction at a room temperature but at a boiling temperature (70-85° bath temperature in the case of dimethylaniline) for 24 hours resulted in the demethylation of the methoxyl group neighboring the oxygen bridge composing the diphenyl ether, which was not affected, and dauricine (II) was formed from (I). (II) was confirmed as its perchlorate of m.p. 215-216°. In the case of (III), the material was completely recovered in either of the solvents and it was found that this reagent is completely inactive to (III). The same treatment of 7-phenoxyquinoline (IV) in ether also ended in the recovery of the starting material.
1) Reaction between 2-benzimidazolethiol and alkyl halide was examined and alkyl derivatives of 2-benzimidazolethiols listed in Tables II, III, and IV were synthesized by heating those compounds in pyridine. Some observations were made on the reaction mechanisms. 2) In these reactions, the alkylation was found to proceed approximately step by step and it was found that the order of the reactivity of each atomic group against alkylation agent, i.e. electrophilic reagent, in the 2-benzimidazolethiol derivatives to be -SH>-N=(ring nitrogen)>-NR- (ring nitrogen).
Two kinds of steroidal sapogenins were isolated from the bulb of Polianthes tuberosa L. (Amaryllidaceae). One is the chief component, melts at 265°, and gives an acetate of m.p. 243-245°, and a 2, 4-dinitrophenylhydrazone of m.p. 275°. Admixture of the genin and its acetate respectively with hecogenin and its acetate showed no depression of the melting point. Further, its chemical properties, analytical values, and infrared absorption spectrum all agreed with those of hecogenin. The other, obtained in a small amount, showed m.p. 207° and the admixture of the genin and its acetate with tigogenin and its acetate showed no depression of the melting point.
From the crude drug, Chih-mu, the rhizome of Anemarrhena asphodeloides Bunge, sarsasapogenin (I) was obtained as the chief component, with a small amount of crystals (II) of m.p. 242-243°. The analytical values of (II) indicated the substance to correspond to the formula of C27H42o4 or C27H44O4, but there is no compound corresponding to it in the known sapogenins. From the rhizome of this plant collected during the winter, only (I) was obtained while the crystals (III) of m.p. 246-248° alone were obtained from its flowers. From the rhizome collected during the summer, the chief component, (I), and crystals (IV) of m.p. 258° were isolated. (IV) formed an acetate of m.p. 185°, and from the analytical values and infrared absorption spetra of the genin and its acetate, (IV) was proved to be identical with markogenin, 22b-spirostan-2ξ, 3β-diol, C27H44O4, isolated from yucca sp. and reported as a new substance by Wall, et al., in 1953. It was also found that (II) and (III) were both impure substances.
The reaction between thiol-type thiamine and cystine (or oxidized-type glutathione) and between thiamine disulfide and cysteine (or reduced-type glutathione) was examined by paper partition chromatography. Both of these reactions showed the same chromatogram and the formation of thiamine-cysteine (or thiamine-glutathione) was confirmed. The former reaction is likely to take place in a living body and may give a suggestion as to the action mechanism of thiamine in a human body.
1) Condensation of 2-hydroxy-4-phenyl-5-aminothiazole and benzaldehyde yielded the 5-benzalamino derivative, which was also obtained by the reaction of α-aminophenylacetonitrile, benzaldehyde, and carbonyl sulfide. Use of p-acetaminobenzaldehyde in place of benzaldehyde in this case results the formation of the 5-p-acetaminobenzalamino derivative. A series of 2-alkoxy or -allyloxy derivatives were prepared by the application of alkyl halide or allyl bromide to the sodium salt of these Schiff's bases. 2) 2-Mercapto-4-methyl-5-benzalaminothiazole was obtained by the reaction of α-aminopropionitrile, benzaldehye, and carbon disulfide. 3) Application of benzaldehyde or p-acetaminobenzaldehyde to 2-mercapto-4-phenyl-5-aminothiazole yielded the corresponding Schiff's bases which were converted to the 2-alkyl (or allyl) thiol derivatives.
1) Side-reactions that occur during oxidation of acylamino compounds with hydrogen peroxide was examined and the chief side-reaction was found to be the formation of ammonia. The cause of this reaction was assumed. 2) The yield of the nitro compound was found to decrease markedly when glacial acetic acid is not used as the solvent. In this case, the amine formed by the hydrolysis of the acetamino group was isolated and it was assumed that hydrogen peroxide also caused hydrolysis.
Negative correlationship was found to exist between the weight and content of the chief component in the thiamine tablets manufactured. This relationship exists due to the correlative state of the granular size and thiamine content of such tablets, which is further influenced by the thiamine-retaining power of various bulking agents, chiefly starch and lactose in the present case, and working conditions.
The die pressure of various tablets coming out of the tabletting machine was measured by the electric resistance strain gauge and the following results were obtained. 1) The tabletting pressure for the tablets of the size and kinds used in the present experiment is in ton units, much larger than was anticipated. 2) With a tabletting machine such as the present one, with movable upper die and stationary lower die, the tablet receives different pressures from either side.
Equilibrium diagram for the diethylamine-water-sodium hydroxide system at 0°, 20°, and 40° was obtained. Diethylamine and water form a completely miscible solution but addition of sodium hydroxide results in partially miscible two liquid phases. Their mutual solubilities decrease with the increase of temperature so that the dehydration effect of sodium hydroxide becomes greater as the temperature increases. In the system of equal composition, the concentration of diethylamine in the upper layer becomes higher with the rise in the temperature. The composition of conjugate solutions of over ten pairs at 0°, 20°, and 40° was determined. According to this result sodium hydroxide does not dissolve into the upper diethylamine layer when the concentration of sodium hydroxide reaches a certain level and diethylamine does not dissolve into the lower sodium hydroxide layer. The concentration of diethylamine present with the saturated solution of sodium hydroxide is approximately 100%. It has become possible, by the utilization of binodal curves and tie-lines of a three-component system, to find the most advantageous method for the dehydration or isolation of organic amines.
Oxidation of L-fucitol with platinum catalyst gave L-fuconic acid only in a maximum of 10% yield and the remainder was found to have decomposed into oxalic and tartronic acids. The same oxidation of D-glucoxe gives D-gluconic acid in 30% yield, about the same result obtained with D-sorbitol.
A kind of cardiotonic glycoside of m.p. 226-228° (decomp.) was isolated in 0.013% yield from the root of Digitalis purpurea L. This substance was found to be identical with digitalinum verum, one of the cardiotonic glycosides present in the leaves and seeds of Digitalis purpurea L.
Microdetermination of acetyl or benzoyl in organic compounds was attempted with the use of a cation exchange resin. The samples were saponified with 5N ethanolic sodium hydroxide solution, passed through a resin bed of Amberlite IR-120, and the amount of acetic acid or benzoic acid in the effluent was titrated with 0.01N sodium hydroxide. The determination was made within an error of ±0.5%. The present analytical procedures are very simple, do not necessitate any special apparatus, take comparatively short time, and good results can be obtained.
The effect of the addition of nonionic surface active agents, such as Tween 80, to the halide solutions in the silver titration with an adsorption indicator, was examined. It was found that the presence of such surfactants prevented the precipitation of silver halides during the course of titration and the solution remained colloidal and clear. The color change of the indicator was sharp and clear as that in acid-alkali titration and the end-point was observed much more easily. The presence of interfering electrolytes, such as sodium sulfate, failed to reduce the accuracy of this titration by the action of the surface active agents.
Bases of aconite roots were extracted in a good yield and in a comparatively short period by continuous extraction using cation exchange resin, Amberlite IRC-50, the procedures being more simple than the existing extraction methods.