Biological change of mesoxalic acid was examined. The paper chromatography of the urine of a dog given mesoxalic acid revealed the presence of mesoxalic acid and a small amount of tartronic acid. Action of the mouse liver homogenate on mesoxalic acid was also examined and it was found to effect reduction of mesoxalic acid to tartronic acid under anaerobic conditions. In order to make a quantitative confirmation of this reduction, mesoxalic acid was injected into a rat and its urine was examined. The urine of a dog given calcium and sodium mesoxalate was also examined and it was found that the substances excreted differed in accordance with the conditions of administration (oral or parenteral) and with calcium and sodium salts. Further decomposition of tartronic acid will be examined in future.
Benzyl cyanide was condensed with dialkylaminoethyl chloride (alkyl=methyl or ethyl) or piperidylethyl chloride in toluene, in the presence of sodium amide, and 2-phenyl-4-dialkylamino (or piperidyl) butyronitrile was obtained. Further condensation of these compounds with 4-chloro-2, 6-lutidine, in the presence of sodium amide, afforded the corresponding 4-lutidyl derivatives. Grignard reaction of such derivatives with ethylmagnesium bromide or methylmagnesium iodide easily afforded the ketone compounds. Reaction of 2-phenyl-2-[4′-(2′, 6′-lutidyl)]-4-dimethylaminobutyronitrile and ethylenediamine gave a compound corresponding to an imidazolyl derivative.
Extract of crude drugs containing biotin-like factors and crystalline biotin dissolved in some kind of solvents afford two separate spots by paper chromatography and they were termed A and B factors. It was assumed from the reports of Melville and Wright that they were respectively biotin and biotin d-sulfoxide. In what form these factors were present in crude drugs and other natural substances was examined by microbiological assay and it was found that biotin d-sulfoxide was not contained in the ligusticum root but both biotin and its d-sulfoxide were present in Liquorice root. Biotin obtained from the extracts of ligusticum and liquorice produced a larger amount of biotin d-sulfoxide than crystalline biotin, the tendency being especially marked in liquorice. The biotin-like factor in Royal jelly, which is reported to contain extremely large amount of biotin, was examined by microbiological assay and the factor was found to be active against Lactobacillus arabinosus but not to Lact. cascei. It was thereby assumed that the biotin-like factor in the Royal jelly is not biotin but biotin d-sulfoxide but this point requires further detailed study.
1) Seasonal variation of the santonin content in the grown-up plants (second, third, fourth, and fifth year growths) of Artemisia kurramensis Qaz. was followed during July 1 to August 21, 1954 (cf. Table I). In these grown-up plants, the maximum in santonin content was reached during the first half of August, being one month later than the usual. This is assumed to have been due to the exceptionally low temperatures prevailing during July, 1954, and the growth had been weak during this period. The atmospheric temperature rose during August to the normal level and the growth became normal. 2) In all first, second, and third year growths of these plants, the growth and development, yield of crop, and santonin content were better in the acclimatized plot than those in the Pakistan origin plot (cf. Tables I, II, and III). The percentage of survived plants after wintering was also higher as the generation of acclimatization was higher. 3) Seasonal variation of the santonin content in the third, fourth, and fifth year growths was approximately of the same tendency as that of the second year growth the content itself being not much different from that of the second year growth. 4) In the experimental plots of the first year growth, the highest santonin content was observed in the first generation of acclimatization by artificial crossing between plants of higher santonin content (cf. Table II). This indicates the possibility of breeding by crossing. 5) Santonin content was analyzed individually in 100 plants of the second year growth and it was found that the range of distribution was 0.00-3.00%, average, 1.486±0.052%, and the mode was 1.00-1.25% (25% of the total) (cf. Table IV). This indicates the necessity and desirability of improving the plants through breeding by clonal separation. 6) Some knowledges were gained regarding the season for carrying out the cutting. 7) It was confirmed that this Artemisia is a short-day plant. 8) The optimum soil moisture for growth seems to lie in the range of 70-90% water capacity of the soil. 9) The santonin content tended to be higher in the plants grown in the soil whose pH was on the alkaline side (pH 7.0-8.5), rather than on the acid side. However, it was observed that this plant grew well in a wide range of pH (3.5-8.5) of the soil.
Some time ago, Ueno and others obtained alkylpyrrolidine by the reduction of N-methyldihydrokaininediol or N-methyldihydrokainine dichloride, derived from kainic acid, and from the results of its Hofmann degradation and other reactions, assumed the structure of 1, 2-dimethyl-3-ethyl-4-isopropylpyrrolidine for the alkylpyrrolidine. This compound was prepared and its first-stage Hofmann degradation and reduction of the methine thereby obtained afforded a dihydromethine compound, possessing only one asymmetric carbon. The infrared absorption spectra of its methiodide, m.p. 186-187°, and that of dihydromethine methiodide, m.p. 207°, [α]D18: -11.5±0.5° (c=1, MeOH), derived from kainic acid, measured in chloroform, agreed in every respect. This has confirmed the alkylpyrrolidine structure and also offered one evidence for the appropriateness of the planar structure of kainic acid proposed earlier.
Pharmacological activity in general of three kinds of safrole derivatives of imidazole and thiazole series was examined and it was found that they possessed only a weak antiepinephrine action. Antispasmodic action of 2-(β-diethylaminoethylamino)-4-(3′, 4′-methylenedioxyphenyl)-5-methylthiazole to excised intestines of a rabbit corresponded to about 4/5 of papaverine and the action was stronger in its methobromide. These compounds also had the action of contracting excised uterine muscle of a guinea pig.
Antispasmodic action of six kinds of safrole derivatives of isoquinoline series, 1-β-dialkylaminoethoxyphenyl-3-methyl-6, 7-methylenedioxyisoquinoline, against smooth muscles was examined. A compound possessing a diethylaminoethoxyl group in the meta-position of the 1-phenyl radical had the strongest action and its effect was somewhat similar to papaverine. This compound also possessed paralytic action on peripheral distribution of the parasympathetic nerves, though much weaker than that of atropine, and its toxicity, LD50, was 0.25mg./g. by subcutaneous injection in mice. Its methobromide was much stronger than the original compound, possessed a weak antihistaminic action, and its toxicity, LD50, was about 0.02mg./g.
1) The strength of antispasmodic action on smooth muscles and toxicity of 1-β-diethylaminoethoxyphenyl-3-methyl-6, 7-methylenedioxyisoquinoline (I), derived from safrole, and its 3, 4-dihydro compound (II) were compared from which (II) was found to have weaker antispasmodic action but there was no significant difference between the toxicity of the two compounds. Analgesic action, thought to be the secondary symptoms of antispasmodic action, was examined in (II) by the modified Hardy method and an effect somewhat stronger than Pethidine Hydrochloride, by the same dose, was observed. 2) 2-Methoxy-6-allyl-1-phenoxyethyldiallylamine, similar to (I) in chemical structure, possessed a strong action of uterine contraction and increased the tonus of smooth muscles. Its methiodide possesses the action of blocking the autonomous nerves but lost the uterine contracting action of the original compound, while it showed antispasmodic action similar to papaverine against the smooth muscles, i.e. actions similar to the methobromide of (I). 3) Relationship between the chemical structure and pharmacological activities of safrole derivatives of isoquinoline, thiazole, and imidazole series was examined.
Relationship between the yield and the heating time, kinds of bases used, and its amount, during the preparation of coumarin-3-carboxylic acid by the condensation of salicylaldehyde and malonic acid were examined and the results obtained are shown in Tables II and III. It was thereby found that (1) the starting material should be mixed while hot, (2) as for the kind and amount of the base used, the yield was better when the ratio of salicylaldehyde: malonic acid: base was 1:2:1 with primary bases and 1:1:2 with secondary bases, while the yield was invariably poor with tertiary bases, and (3) aniline proved to be the best of the bases used on the yield. From the reaction of 2-hydroxybenzylidene-aniline, -p-toluidine, and -bispiperidine, this condensation reaction was found to require a 2-hydroxybenzylidene base as the intermediate.
Reaction conditions during the preparation of 6-nitrocoumarin-3-carboxylic acid from 5-nitrosalicylaldehyde were examined and results shown in Tables I and II were obtained. It was found that (1) there was marked effect from the time of heating, differing from the case of salicylaldehyde, (2) the yield was better when the amount of aniline added was less than 0.5 mole of the amount of malonic acid, though the effect was not as marked as in the case of salicylaldehyde, and (3) from these reactions, this condensation was also found to pass through the intermediate of 5-nitro-2-hydroxybenzylideneaniline which underwent condensation by the action of an excess of malonic acid or hydrochloric acid. These conditions were utilized in preparing nitrocoumarin-3-carboxylic acids with a nitro group in 5-, 7-, or 8-position, and their ethyl esters.
2-Hydroxy-5-Acetylaminobenzylideneaniline (I) was prepared by the reduction of 5-acetylaminosalicylic acid with sodium amalgam, in the presence of aniline, and (I) was derived to 6-acetylaminocoumarin-3-carboxylic acid (II), its ethyl ester (IV), and 6-aminocoumarin-3-carboxylic acid (III) and its ethyl ester (V) (cf. Fig. 1). The products obtained by the catalytic and chemical reduction of 6-nitrocoumarin-3-carboxylic acid and its ethyl ester were confirmed to be (III) and (V). 4-Aminosalicylic acid was derived, similarly as the 6-amino derivative, to 7-acetylaminocoumarin-3-carboxylic acid (VII) and its ethyl ester (IX), and 7-aminocoumarin-3-carboxylic acid (VIII) and its ethyl ester (X) (cf. Fig. 2). It was observed that the reduction of 7-nitrocoumarin-3-carboxylic acid and its ethyl ester also yielded the 7-amino derivatives. Both the 7-amino and 7-acetylamino derivatives thereby obtained possessed a strong fluorescence.
Soporific and sedative actions and toxicity of 25 kinds of coumarin-3-carboxylic acid derivatives were examined in mice and the results obtained are listed in the accompanying Tables and Figures. 1) Esters possess some efficacy and the effect is stronger in alkyl esters than in phenyl esters. 2) In alkly esters, the activity became weaker with the increase in the size of the alkyl group and in the same alkyl group, the normal chain was weaker than the iso chain. 3) Compounds of -CONHR type where the R is an aromatic ring or ureido compounds were ineffective and devoid of toxicity. 4) Substitution of coumarin ring in the 6-position with nitro, amino, or acetylamino group caused loss of efficacy and toxicity. 5) The same substitution in the 7-position of the coumarin ring also caused loss of the sedative and soporific activities but not of toxicity, which appeared gradually over a few days.
Ethyl 7-nitrocoumarin-3-carboxylate and ethyl coumarin-3-carboxylate were each injected into a mouse or applied on excised liver and kidney of a mouse and the fluorescent substance found in the urine, liver, and kidneys was examined by paper chromatography. The results shown in Tables I, II, and IV, suggested the in vitro change of these compounds to be as follows: 1) Ethyl 7-nitrocoumarin-3-carboxylate is chiefly reduced in the liver and the kidney and hydrolyzed at the same time to 7-aminocoumarin-3-carboxylic acid. A part of it is further acetylated in the liver to the 7-acetylamino derivative, these two substances being excreted in the urine. 2) Ethyl coumarin-3-carboxylate undergoes hydrolysis and other changes in the liver and a part of it changes to coumarin-3-carboxylic acid.
Determination of sodium ion by flame photometry by direct ignition of hot spring and mineral spring water, using the standard calibration curve of standard sodium chloride solution gives negative error which cannot be disregarded, with the exception of a common salt spring. Good analytical result can be obtained if the spring water is treated at pH 4 to remove carbon dioxide and hydrogen sulfide, passed through an R-Cl type column of ion exchange resin (Amberlite IRA-410), thereby changing all the anions in the sample water to chlorine ion, and finally submitted to flame photometry. This method can be applied widely to the majority of neutral salt springs in Japan, excluding the acid, alum, and acid alum-vitriol springs. It is better than the chemical determination of sodium in that it gives better accuracy and the shorter time required. The weak acid components which cannot be substituted by the foregoing resin can be disregarded.
It has been revealed that the catalytic vapor phase reaction of allyl alcohol, ketones, and ammonia afforded pyridine bases with a substituent in 2 or in 2 and 3 positions. 2-Picoline and 2-phenylpyridine were obtained respectively from acetone and acetophenone, 2, 3-dimethylpyridine from methyl ethyl ketone, and 2-methyl-3-ethylpyridine from methyl propyl ketone. Reaction of diethyl ketone, allyl alcohol, and ammonia gave 2-ethyl-3-methylpyridine. Of the catalysts used, cadmium phosphate on activated white clay gave the best results, while the optimal reaction temperature was 350-400°. Use of about 3moles of ammonia to 1 mole each of allyl alcohol and the ketone afforded the desired base in 20-30% yield.
2-Methyl-3-alkylpyridines were obtained by the catalytic vapor phase reaction of allyl alcohol, methyl alkyl ketone, and ammonia, using cadmium phosphate-acid clay catalyst. The use of ethyl as the alkyl in the ketone afforded 2, 3-lutidine (yield, 24%), propyl gave 2-methyl-3-ethylpyridine (yield, 16%), butyl gave 2-methyl-3-propylpyridine (yield, 15%), and amyl gave 2-methyl-3-butylpyridine (yield, 18%), with minute amounts of 3-picoline and 3, 5-lutidine as by-products. 2-Alkylpyridine was not detected at all from this reaction. Reaction temperature was 375-425° and the molar ratio of 1-1.5:1 for allyl alcohol to methyl alkyl ketone, and 3-6 times the theoretical amount of ammonia were used. When the amount of allyl alcohol was larger, the yield of 2-methyl-3-alkylpyridine became greater. Cadmium tungstate or chromate also was good and could be used as a catalyst.
1) Testosterone and its esters show sensitive blue coloration with 0.1% ferric chloride in 70% sulfuric acid and 3% acetic acid, up to 0.2mg. 2) This coloration is extremely stable compared to coloration reactions of steroids in general. 3) Testosterone and its esters, within a definite concentration range, show linearity and parallelism in their coloration, which are accurate and can be used for colorimetry. 4) Application of this colorimetry to commercial males hormone injections gave much more simple and accurate means of determination than the existing bioassay and the fiducial limit of error was found to be in 90-110% range.
Root powder of Aristolochia debilis was extracted with benzene and the crystalline component thereby separated was designated aristolone. Aristolone, m.p. 101°, C15H24O, is a saturated sesquiterpene ketone, gives negative iodoform reaction, and does not form isonitroso or benzylidene compound. From its analytical data, aristolone must be tricyclic. Aristolone does not give naphthalene derivative when heated with selenium. Isoaristolone, obtained by treating aristolone with sulfuric tricyclie acid, was reduced with lithium aluminum hydride aud then heated with selenium, bywhich a hydrocarbon, aristolin, C15H18, was obtained. Its picrate, m.p. 135.5°, is a new naphthalene derivative. From its behavior to selenium, sulfuric acid, and sodium amide, and the fact that it is a six-membered cyclic ketone, its structural formula was assumed to be either A, B, or C (XVII).
A simple, wide-rage millicoulometer, utilizing blocking oscillator, was made on trial. By utilizing the change in electric pressure by the current flowing through the resistance, Rg (Fig. 1), in the grid circuit, the resistance in the resistance circuit containing 65J7-GT tube was made to change automatically and the oscillating frequency was made proportional to the currency. The linearity was as shown in Fig. 3. Reproducibility was within 0.2%. Controlled potential coulometric titration of microgram amounts of copper, lead, and cadmium was carried out with this coulometer. If the background count is made definite and the ratio of that to the total count made as small as possible, copper can be determined within an error of 1μg. (cf. Table I). Determination of a mixture of lead and cadmium gave larger error, as shown in Table II.
Titration of carboxylic acids in nonaqueous solution sometimes gives an N-shaped characteristic curve. This was proved to be due to the kind of the solvent used, or rather to the value of the dielectric constant of the solution, by examining the matter with methanol-benzene system and other mixed solvent systems. The distance between the carboxyl groups in each acid defines the dielectric constant (higher value), Dc, of the limit at which an N-shaped curve appears. By the application of the Coulomb's law to the mutual action of the carboxyl groups, and when its force f is fc at Dc, fc becomes a common constant for all the carboxylic acids, and the equations (3) and (4) are obtained. From these equations, K is obtained with acetylenedicarboxylic acid as the standard, and interpolation of the value of r enables calculation of the value of Dc. Values thereby obtained agree well with experimental values (cf. Table I). This has proved that the formation of an N-shaped curve is chiefly due to the Coulomb force between the carboxyl groups fortified in a solvent with a small dielectric constant.
Effect of cadaverine, putrescine, agmatine, ethylendiamine, ethanolamine, histamine, tyramine, choline, γ-aminobutyric acid, and δ-aminovaleric acid on the riboflavin production by Eremothecium ashbyii was examined with synthetic medium. Addition of 10-5 mole of cadaverine, putrescine, and ethylenediamine effected 72%, 21%, and 37% respective increase in riboflavin production, compared to the control without any addition. Agmatine, histamine, and tyramine in 10-3 mole concentration clearly injured riboflavin production but other amines were ineffective (cf. Tables II to XII).
Surface area and acetylene adsorptive ability of various zinc oxide catalysts of different make were measured. It was found that in adsorption of acetylene, Van der Waal's adsorption occurred at temperatures below 80°, and chiefly activated adsorption, accompanied with partial polymerization and decomposition, occurred at temperatures above 80°C. The physical adsorption of acetylene was found to be the maximum in a catalyst prepared by the pyrolysis of basic zinc carbonate precipitated with sodium carbonate. The same results were obtained with the surface area of the catalyst. Activated adsorption was the greatest in a catalyst prepared by the use of ammonium carbonate as the precipitant of basic zinc carbonate. As for the effect of sodium as an impurity and the temperature of heating, sodium was found to reduce both the amount of activated adsorption of acetylene and surface area of the catalyst. The surface area and acetylene adsorption were also found to be markedly decreased by heating zinc oxide to 500°C.
There is a possibility of a formation of various cyclization products by the application of monochloroacetic acid to potassium 3-benzoyldithiocarbazate (VI). Actually, two products of m.p. 174° and m.p. 170° are obtained and from various reactions, the former was found to be 3-benzoylaminorhodanine (X) and the latter an oxadiazole derivative (XV). (X) undergoes condensation with benzaldehyde to give a benzal compound (XX) but transits to a compound of m.p. 184° by treatment with hydrochloric acid, which was confirmed to be a thiadiazole derivative (XI). Application of ethyl monochloroacetate to (VI) yields an oily substance from which an oxadiazole derivative (XIV) and a thiadiazine derivative (XXV) were obtained.
Bond energy and entropy changes of the hydrogen bonding between quinoline 1-oxide and phenol and various phenol derivatives were examined by the measurements of near-ultraviolet absorption and following considerations were made from the values thereby obtained: 1) Hydrogen-bonding ability of phenol derivatives was examined with o-, m-, and p-cresols, o-, m- and p-chlorophenols, and guaiacol. 2) Strength of quinoline or pyridine 1-oxide as a proton acceptor was examined. 3) Strength of hydrogen bonding between N-oxides and phenols or alcohols was calculated, using an electrostatic model and the measured values were compared. 4) Ability to form hydrogen bonding in excited states was examined with theforegoing systems and it was found that the hydrogen bonding of aromatic N-oxides became weaker in an excited state. 5) It is assumed that the contribution of molecular compound formation to the energy of these kinds of hydrogen bonding is extremely small.
Dissociation constant, pKa, of pyridine and quinoline 1-oxides was determined spectrochemically and the respective values of 0.56±0.01 and 0.70±0.07 were obtained. These values are far smaller than those (4.04-4.96) of aliphatic, tertiary amine N-oxides. This was assumed to be chiefly due to the resonance of heterocyclic N-oxides as shown in (II) and its contribution was calculated to be 5.2-5.6kcal./mole from equation (4). These values agree well with the tendency of the shift of absorption spectra. Dissociation constant becomes larger when an amino group is introduced into the 4-position and this was assumed to be due to the contribution of an ionic resonance structure such as shown in (V). The fact was comparatively examind with pyridine derivatives.
When diethyl crotonoylmalonate is treated under conditions for the Michael condensation, (1) diethyl β-methyl-γ-ethoxycarbonylglutarate is formed in the case of sodium ethoxide in ethanol, and dimethyl β-methyl-γ-methoxycarbonylglutarate with sodium methoxide in a large amount of methanol, (2) in the case of tert-butoxide in tert-butanol alcoholysis does not occur and there is no formation of the foregoing glutarate, and (3) in the case of piperidine, β-(1-piperidyl) butyrate is formed.
Treatment of diethyl crotonoylmalonate under conditions for the Michael condensation does not essentially cause alcoholysis when alcohols are not present, and (1) the starting material is recovered in the case of piperidine and (2) the intermolecular Michael condensation occurs when sodium ethoxide is used in benzene to form a cycloöctanedione derivative (assumed).
Difference in the ability to absorb carbon dioxide by the anhydride and hydrate of diphenylhydantoin sodium was studied using the constant-pressure adsorption measuring apparatus. It was found that whereas the anhydride did not absorb dry carbon dioxide, the hydrate absorbed carbon dioxide, slowly at first and then rapidly, about 1 mole of carbon dioxide per mole of hydantoin sodium, liberating diphenylhydantoin and sodium hydrogen carbonate. Absorption of carbon dioxide by the hydrate was shown to become more active as the moisture and temperature of the vapor phase and partial pressure of carbon dioxide became higher.
For comparison with mono-β-chloroethylsuifonyl compounds, bis (β-chloroethylsulfonyl) derivatives of ethane (1, 2), benzene (m, p), toluene (3, 4), and naphthalene (1, 5) were prepared and their bacteriostatic activity in vitro against Myco. tuberculosis, Staph. aureus, and E. coli was tested (Table I). These bifunctional compounds showed, contrary to expectations, almost the same grade of antibacterial activity as the monofunctional ones. It was clarified that bis (β-chloroethylsulfonyl) group was easily converted to bis (vinylsulfonyl) group in sodium hydrogen carbonate buffer solution and moreover, the latter reacted with many nucleophilic reagents, such as mercaptans and amines, under the formation of bis-β-substituted derivatives.
Two kinds of new substances, named justcin and isojusticin, were obtained as crystals melting respectively at 216° and 251-252°, from the whole herb of Justicia procumbens L. var. leucantha Honda, in 0.1% and 0.01% yield. Both compounds agreed with the formula C16H12O5, possessed one lactone ring and two methoxyl groups, and the nature of the lactone ring was not that of ordinary coumarins. The same treatment of Dicliptera japonica Makino, Hygrophila lancea Miq., and Acanthus mellis L., all belonging to the same Acanthaceae family, failed to afford any compounds similar to justicin.
Gein was detected from Geum jeponicum. Phenolic glycosides and sugars in allied plants, Geum aleppicum var. sachalinense, Pavageum calthifolium, Potentila fragarioides var. Sparengelliana, P. Freyniana, and Agrimonia Eupatria were detected by paper chromatography.
Triethyl butylcitrate (IV) was prepared from diethyl oxalylcaproate and ethyl bromoacetate, and triethyl tetradecylcitrate (V) from triethyl oxalylpamitate and ethyl iodoacetate. Saponification of (V) with alkali hydroxide afforded palmitic acid and tetradecylcitric acid (VII).